Bidirectional multi-enzymatic scaffolds for biosynthesizing cannabinoids

BIDIRECTIONAL MULTI-ENZYMATIC SCAFFOLDS FOR BIOSYNTHESIZING CANNABINOIDS

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Application Serial Nos. 62/836,265, filed on April 19, 2019 and 62/771,839, filed on November 27, 2018. The disclosures of the prior applications are considered part of the disclosure of this application, and are incorporated in their entirety into this application.

TECHNICAL FIELD

This document relates to methods and materials for biosynthesizing cannabinoids, and more particularly to using bidirectional multi-enzymatic scaffolds to biosynthesize cannabinoids.

BACKGROUND

The emerging therapeutic potential of cannabinoids warrants industrial-scale production to meet compounding future demands. Traditional cannabinoid production efforts rely on large-scale farming of Cannabis sativaL. However, agricultural cannabinoid production is problematic due to issues such as uncontrollable environmental factors and scaling limitations.

SUMMARY

This document is based, at least in part, on the discovery that a bidirectional, multi-enzymatic scaffold can be engineered to allow high-throughput cannabinoid production in recombinant host cells. By controlling the localization, spatial orientation, and stoichiometry of enzymes catalyzing the biosynthesis of cannabinoids and cannabinoid precursors, the multi-enzymatic scaffolds described herein allow fluxoptimized cannabinoid biosynthesis in genetically-engineered host cells.

In one aspect, this document features a host cell capable of producing one or more cannabinoids selected from the group consisting of cannabigerolic acid, cannabidiolic acid, and cannabichromenic acid. The host cell includes at least three different exogenous 1

nucleic acids, wherein the first and the second exogenous nucleic acids each encode a plurality of engineered enzymes selected from the group consisting of acetyl-CoA acetyltransferase, a 3-hydroxybutyiyl-CoA dehydrogenase, an enoyl-CoA hydratase, a beto-ketothiolase, a trans-enoyl-CoA reductase, an HMG-C0A synthetase, an HMG-C0A reductase, a mevalonate kinase, a phosphomevalonate kinase, a diphosphomevalonate decarboxylase, an isopentenyl-diphosphate delta isomerase, a geranyl-diphosphate synthase, an olivetol synthase, an olivetolic acid cyclase, and a CBGA synthase; wherein each of the engineered enzymes includes a heterologous interaction domain, wherein the heterologous interaction domain comprises a first and a second peptide motif, and wherein each heterologous interaction domain is different from each other; and wherein the third exogenous nucleic acid encodes a polypeptide scaffold comprising a plurality of peptide ligands, wherein each peptide ligand comprises an amino acid sequence that can bind to the first or the second peptide motif of one of the heterologous interaction domains. The plurality of engineered enzymes further can include an ATP citrate lyase and an acetyl-CoA carboxylase. The host cell further can include an exogenous nucleic acid encoding a cannabidiolic acid synthase (CBDAS) and a cannabichromenic acid synthase (CBCAS). The host cell can include an exogenous CBDAS. The host cell can include an exogneous CBCAS. The host cell can include an exogenous CBDAS and an exogenous CBCAS. The host cell can include an exogenous hexanoyl-CoA synthetase. The host cell can include at least four different exogenous nucleic acids, wherein the first, second, and fourth nucleic acids each encode a plurality of the engineered enzymes. The host cell can include at least five different exogenous nucleic acids, wherein the first, second, fourth, and fifth nucleic acid each encode a plurality of the engineered enzymes. The host cell can include at least six different exogenous nucleic acids, wherein the first, second, fourth, fifth, and sixth nucleic acids each encode a plurality of the engineered enzymes. Each exogenous nucleic acid can include a constitutive promoter operably linked to the sequence encoding the engineered enzyme or polypeptide scaffold or an inducible promoter operably linked to the sequence encoding the engineered enzyme or polypeptide scaffold. In some embodiments, the promoter is a GALI-10 promoter. In some embodiments, a constitutive promoter used to express the polypeptide scaffold has

weaker constitutive activity level than a constitutive promoter used to express the engineered enzymes. In some embodiments, a constitutive promoter is used to express the engineered enzymes and an inducible promoter is used to express the polypeptide scaffold. In some embodiments, an inducible promoter is used to express the engineered enzymes and a constitutive promoter is used to express the polypeptide scaffold.

Any of the host cells can be bacterial, yeast, algae, or plant cells. Abacterial cell can be selected from the group consisting of Escherichia coh, Bacillus, Brevibacterium, Streptomyces, and Pseudomonas cells. A yeast cell can be selected from the group consisting of Pichiapastoris, Saccharomyces cerevisiae, Yarrowia lipolytica, Kluyveromyces marxianus, and Komagataellaphaffii cells. An algae cell can be Dunaliella sp., Chlor ella variabilis, Euglena mutabilis, or Chlamydomonas reinhardtii cells. A plant cell can be a Cannabis or tobacco cell.

In some embodiments, each of the engineered enzymes is of the formula: enzyme - linkeri - spacer - linker2 - motifi - linkers - motif2, where linkers 1, 2, and 3 can be the same or different, motif 1 and motif 2 can be the same or different, and where motif 1 and motif 2 form the heterologous interaction domain. A scaffold polypeptide can be of the formula: N-terminus-[Ligand 1 - linker - Ligand 2 -Spacer]n -(optionally-tagged) Cterminus, where n is the number of heterologous interaction domains, and where ligand 1 and ligand 2 bind motif 1 and motif 2, respectively, of the heterologous interaction domain. The scaffold polypeptide can be tagged with a MYC tag, FLAG tag, or HA tag. The host cell further can include a nucleic acid encoding a second polypeptide scaffold comprising a plurality of peptide ligands, wherein each peptide ligand comprises an amino acid sequence that can bind to a different motif of the heterologous interaction domain. The linker can have a flexible GS-rich sequence flanking a rigid a-helical moiety. The spacer can be the cTPR6 spacer.

This document also features a method of producing one or more cannabinoids selected from the group consisting of cannabigerolic acid, cannabidiolic acid, and cannabichromenic acid. The method can include culturing any of the host cells described herein under conditions wherein the host cell produces the one or more cannabinoids. The host cells can be cultured in a culture medium supplemented with citrate, glucose,

hexanoic acid, and/or other carbon source, and/or in a culture medium supplemented with malonyl-CoA. The method further can include extracting the one or more cannabinoids from the host cells.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Other features and advantages of the invention will be apparent from the following detailed description, and from the claims.

DESCRIPTION OF DRAWINGS

FIG lAis a schematic of one representative embodiment of a multi-enzymatic cannabinoidergic scaffold within a cell. The multi-enzymatic scaffold includes enzymes of the hexanoyl-CoA pathway, enzymes of the upper cannabinoid pathway, and enzymes of the mevalonate pathway. The schematic also depicts a second scaffold according to one embodiment containing enzymes of the malonyl-CoA pathway and depicts a nonscaffolded cannabidiolic acid synthase (CBDAS) and a non-scaffolded cannabichromenic acid synthase (CBCAS). ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzyme-linked ID. The target products cannabigerolic acid (CBGA), cannabigerol (CBG), cannabidiolic acid (CBDA), cannabidiol (CBD), cannabichromenic acid (CBCA), and cannabichromene (CBC), are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘A’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. IB is a schematic of one representative embodiment of a bidirectional, multi-enzymatic scaffold within a cell (e.g., a yeast cell). The multi-enzymatic scaffold (referred to as SCF gene cassette in the nucleus) includes enzymes of the hexanoyl-CoA pathway (referred to as HCA cassette in nucleus), enzymes of the upper cannabinoid pathway (referred to as CAN cassette in nucleus), and enzymes of the mevalonate pathway (referred to as GPP cassette in nucleus). The schematic also depicts a second scaffold according to one embodiment containing enzymes of the malonyl-CoA pathway and depicts a non-scaffolded CBDAS and a non-scaffolded CBCAS. ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzyme-linked ID. The target products CBGA, CBG, CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘A’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 2Ais a schematic of gene cassettes according to one embodiment for the engineering of cannabinoidergic cells.

FIG. 2B is a schematic of gene cassettes used in Examples 2-4 for biosynthesizing cannabinoids in yeast.

FIG. 3 is an example of an enzyme-scaffold complex.

FIG. 4 is a schematic of one representative embodiment of a multi-enzymatic cannabinoidergic scaffold within a cell. The multi-enzymatic scaffold includes enzymes of the hexanoyl-CoA pathway, enzymes of the upper cannabinoid pathway, and enzymes of the mevalonate pathway. The schematic also depicts a second scaffold according to one embodiment containing enzymes of the malonyl-CoA pathway and depicts a nonscaffolded CBDAS and a non-scaffolded CBCAS. Pyruvate dehydrogenase (El) and dihydrolipoyl transacetylase (E2) are substituted for ATP citrate lyase in both of the depicted scaffolds. ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzyme-linked ID. The target products CBGA,

CBQ CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and

CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘A’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 5 is a schematic of one representative embodiment of a multi-enzymatic cannabinoidergic scaffold within a cell. The multi-enzymatic scaffold includes enzymes of the hexanoyl-CoA pathway, enzymes of the upper cannabinoid pathway, and enzymes of the MEP (2-C-methylerythritol 4-phosphate) pathway. The schematic also depicts a second scaffold according to one embodiment containing enzymes of the malonyl-CoA pathway and depicts a non-scaffolded CBDAS and a non-scaffolded CBCAS. ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzyme-linked ID. The target products CBGA, CBG; CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘A’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 6A contains the amino acid sequence of each of the following enzymes: an ATP citrate lyase (SEQ ID NO:83), acetyl-CoA acetyltransferase (atoB) (SEQ ID NO:84), a 3-hydroxybutyiyl-CoA dehydrogenase (SEQ ID NO:85), an enoyl-CoA hydratase (SEQ ID NO:86), a trans-enoyl-CoA reductase (SEQ ID NO:88), a betaketothiolase (bktB) (SEQ ID NO:87), an HMG-C0A synthase (SEQ ID NO :90), a truncated HMG-C0A reductase (SEQ ID NO:91), a mevalonate kinase (SEQ ID NO:92), a phosphomevalonate kinase (SEQ ID NO:93), a diphosphomevalonate decarboxylase (SEQ ID NO:94), an isopentenyl-diphosphate delta isomerase (SEQ ID NO:95), a mutant geranyl-diphosphate synthase (ERG20ww) (SEQ ID NO:96), an olivetol synthase (SEQ ID NO:98), an olivetolic acid cyclase (SEQ ID NO:99), a CBGA synthase (SEQ ID NO: 100), an acetyl-CoA carboxylase (SEQ ID NO:97), a CBDA synthase (SEQ ID NO: 101), a CBCA synthase (SEQ ID NO: 102), and a hexanoyl-CoA synthetase (SEQ ID NO:89).

FIG. 6B contains the amino acid sequence of engineered enzymes of the formula Enzyme - Enzyme Linker - cTPR6 Spacer - ID Linker - ID Motif #1 - ID Motif Linker - ID Motif #2, where the linkers (enzyme linker, ID linker, and ID motif linker) can be the same or different, and ID motif #1 and ID motif #2 can be the same or different. The amino acid sequence of the following engineered enzymes are provided: ATP citrate lyase (ID1) (SEQ ID NO: 103), an acetyl-CoA acetyltransferase (atoB) (ID2) (SEQ ID NO: 104), a 3-hydroxybutyryl-CoA dehydrogenase (ID3) (SEQ ID NO: 105), an enoylC0A hydratase (ID4) (SEQ ID NO: 106), a trans-enoyl-CoA reductase (ID5) (SEQ ID NO: 107), a beto-ketothiolase (bktB) (ID6) (SEQ ID NO: 108), an HMG-C0A synthase (ID7) (SEQ ID NO: 109), a truncated HMG-C0A reductase (ID8) (SEQ ID NO: 110), a mevalonate kinase (ID9) (SEQ ID NO:111), a phosphomevalonate kinase (ID10) (SEQ ID NO: 112), a diphosphomevalonate decarboxylase (ID11) (SEQ ID NO: 113), an isopentenyl-diphosphate delta isomerase (ID12) (SEQ ID NO: 114), a mutant geranyldiphosphate synthase (ERG20ww) (ID13) (SEQ ID NO:115), an olivetol synthase (ID14) (SEQ ID NO:116), an olivetolic acid cyclase (ID15) (SEQ ID NO:117), a CBGA synthase (ID 16) (SEQ ID NO: 118), and an acetyl-CoA carboxylase (ID 17) (SEQ ID NO:211).

FIG. 6C contains the amino acid sequence of a polypeptide scaffold of the formula: N-terminus - [Ligand #1 - ID Motif #1 Ligand - Linker - ID Motif #2 Ligand Scaffolded ID-binding Site Spacer]n - (Myc)3-tagged C-terminus, where n is 16 and the ID motif ligands correspond to the motifs for IDs 1-16 as shown in Table 2. See SEQ ID NO:119.

FIG. 6D contains the amino acid sequence of a polypeptide scaffold of the formula: N-terminus - [Ligand #1 - ID Motif #1 Ligand - Linker - ID Motif #2 Ligand Scaffolded ID-binding Site Spacer]n - (FLAG)3-tagged C-terminus, where n is 2 and the ID motif ligands correspond to the motifs for IDs 1 and 17 as shown in Table 2. See SEQ ID NO: 120.

FIG. 7 is a schematic of one representative embodiment of a scaffold with the minimal requirements for cannabigerolic acid synthesis. The scaffold contains enzymes of the upper cannabinoid pathway. In this embodiment, a non-scaffolded hexanoyl-CoA

synthetase (HCS), a non-scaffolded CBDAS, and a non-scaffolded CBCAS also are used. ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzyme-linked ID. The target products CBGA, CBG, CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘A’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 8 is a schematic of one representative embodiment of a bi-directional scaffold containing a HCS on the N-terminus of the scaffold, a geranyl pyrophosphate synthase (GPPS) on the C-terminus of the scaffold, and the enzymes of the upper cannabinoid pathway between the HCS and GPPS. In this embodiment, a non-scaffolded CBDAS and a non-scaffolded CBCAS also can be used. ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzymelinked ID. The target products CBGA, CBG; CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘A’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 9 is a schematic of one representative embodiment of a unidirectional scaffold containing enzymes of the upper cannabinoid pathway, shown with soluble enzymes from the precursor pathways (hexanoyl-CoA pathway, mevalonate pathway, and malonyl-CoA pathway), and soluble CBDAS and CBCAS. ID refers to enzyme-linked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzymelinked ID. The target products CBGA, CBG; CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA.

For each decarboxylation, the ‘Δ’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 10 is a schematic of one representative embodiment of a multi-enzymatic cannabinoidergic scaffold within a cell. The multi-enzymatic scaffold includes enzymes of the malonyl-CoA (MCA) pathway, enzymes of the upper cannabinoid pathway, and enzymes of the mevalonate pathway. The schematic also depicts a separate scaffold according to one embodiment containing enzymes of the hexanoyl-CoA pathway and depicts a non-scaffolded CBDAS and a non-scaffolded CBCAS. ID refers to enzymelinked interaction domain; cTPR6 refers to a spacer sequence; scaffolded ligands refer to the tandem peptide ligands that form the scaffold-binding sites specific for each enzymelinked ID. The target products CBGA, CBG; CBDA, CBD, CBCA, and CBC are boxed for emphasis. CBG can be produced by decarboxylation of CBGA, CBD can be produced by decarboxylation of CBDA, and CBC can be produced by decarboxylation of CBCA. For each decarboxylation, the ‘Δ’ symbols represent heat and the ‘hv’ symbols represent light.

FIG. 11 is a schematic of one representative embodiment of a multi-enzymatic cannabinoidergic scaffold within dual compartments of a cell, the cytosol and mitochondri a/pl asti d.

FIG 12A contains the nucleotide sequences encoding each of the following: an ATP citrate lyase (SEQ ID NO: 121), an acetyl-CoA acetyltransferase (atoB) (SEQ ID NO: 122), a 3-hydroxybutyiyl-CoA dehydrogenase (SEQ ID NO: 123), an enoyl-CoA hydratase (SEQ ID NO: 124), a trans-enoyl-CoA reductase (SEQ ID NO: 125), a betoketothiolase (bktB) (SEQ ID NO: 126), an HMG-C0A synthase (SEQ ID NO: 127), a truncated HMG-C0A reductase (SEQ ID NO: 128), a mevalonate kinase (SEQ ID NO: 129), a phosphomevalonate kinase (SEQ ID NO: 130), a diphosphomevalonate decarboxylase (SEQ ID NO: 131), an isopentenyl-diphosphate delta isomerase (SEQ ID NO: 132), a geranyl-diphosphate synthase (ERG20ww) (SEQ ID NO: 133), an olivetol synthase (SEQ ID NO: 134), an olivetolic acid cyclase (SEQ ID NO: 135), a CBGA synthase (SEQ ID NO: 136), an acetyl-CoA carboxylase (SEQ ID NO: 137), a CBDA

synthase (SEQ ID NO: 138), a CBCA synthase (SEQ ID NO: 139), and a hexanoyl-CoA synthetase (SEQ ID NO: 140).

FIG. 12B contains the nucleotide sequences encoding engineered enzymes of the formula: Enzyme - Enzyme Linker - cTPR6 Spacer - ID Linker - ID Motif #1 - ID Motif Linker - ID Motif #2, where the Enzyme Linker, ID Linker, and ID Motif Linker can be the same or different, and where ID Motif #1 and ID Motif #2 can be the same or different. The nucleotide sequences encoding the following engineered enzymes are provided: ATP citrate lyase (ID1) (SEQ ID NO: 141), an acetyl-CoA acetyltransferase (atoB) (ID2) (SEQ ID NO: 142), a 3-hydroxybutyryl-CoA dehydrogenase (ID3) (SEQ ID NO: 143), an enoyl-CoA hydratase (ID4) (SEQ ID NO: 144), a trans-enoyl-CoA reductase (ID5) (SEQ ID NO: 145), a bktB (ID6) (SEQ ID NO: 146), an HMG-C0A synthase (ID7) (SEQ ID NO: 147), a truncated HMG-C0A reductase (ID8) (SEQ ID NO: 148), a mevalonate kinase (ID9) (SEQ ID NO: 149), a phosphomevalonate kinase (ID10) (SEQ ID NO:150), a diphosphomevalonate decarboxylase (ID11) (SEQ ID NO:151), an isopentenyl-diphosphate delta isomerase (ID12) (SEQ ID NO: 152), a mutant geranyldiphosphate synthase (ERG20ww) (ID13) (SEQ ID NO: 153), an olivetol synthase (ID14) (SEQ ID NO: 154), an olivetolic acid cyclase (ID 15) (SEQ ID NO: 155), a CBGA synthase (ID16) (SEQ ID NO: 156), and an acetyl-CoA carboxylase (ID17) (SEQ ID NO: 157).

FIG. 12C contains the nucleotide sequence (SEQ ID NO: 158) encoding a scaffold polypeptide that contains the peptide ligands corresponding to IDs 1-16 as shown in Table 2 and a triplicate myc tag on the C-terminus.

FIG. 12D contains the nucleic acid sequence (SEQ ID NO: 159) encoding a scaffold polypeptide that contains the peptide ligands corresponding to IDs 1 and 17, and a triplicate FLAG tag on the C-terminus.

FIG. 13 A contains the amino acid sequence of scaffold-binding engineered enzymes and a soluble hexanoyl-CoA synthetase (HCS) (SEQ ID NO:209) encoded by the HCA gene cassette. The scaffold-binding engineered enzymes are ATP Citrate Lyase (ACL) (ACL - Enzyme Linker - cTPR6 Spacer - ID Linker - ID1) (SEQ ID NO: 160); Acetyl-CoA Acetyltransferase (atoB) (atoB - Enzyme Linker - cTPR6 Spacer - ID

Linker - ID2) (SEQ ID NO: 161); 3-Hydroxybutyryl-CoA Dehydrogenase (BHBD) (BHBD - Enzyme Linker - cTPR6 Spacer - ID Linker - ID3) (SEQ ID NO: 162); EnoylC0A Hydratase (ECH) (ECH - Enzyme Linker - cTPR6 Spacer - ID Linker - ID4) (SEQ ID NO: 163); Trans-Enoyl-CoAReductase (ECR) (ECR - Enzyme Linker - cTPR6 Spacer - ID Linker - ID5) (SEQ ID NO :164); and Beta-Ketothiolase (bktB) (bktB Enzyme Linker - cTPR6 Spacer - ID Linker - ID6) (SEQ ID NO: 165).

FIG. 13B contains the amino acid sequences of scaffold-binding engineered enzymes encoded by the GPP gene cassette. The scaffold-binding engineered enzymes are HMG-C0A Synthase (HMGS) (HMGS - Enzyme Linker - cTPR6 Spacer - ID Linker - ID7) (SEQ ID NO: 166); truncated HMG-C0A Reductase (tHMGR) (tHMGR Enzyme Linker - cTPR6 Spacer - ID Linker - ID8) (SEQ ID NO: 167); Mevalonate Kinase (ERG12) (ERG12 - Enzyme Linker - cTPR6 Spacer - ID Linker - ID9) (SEQ ID NO: 168); Phosphomevalonate Kinase (ERG8) (ERG8 - Enzyme Linker - cTPR6 Spacer - ID Linker - ID 10) (SEQ ID NO: 169); Diphosphomevalonate Decarboxylase (MVD1) (MVD1 - Enzyme Linker - cTPR6 Spacer - ID Linker - ID11) (SEQ ID NO: 170); Isopentenyl-Diphosphate Delta-Isomerase (IDI1) (IDI1 - Enzyme Linker - cTPR6 Spacer - ID Linker - ID 12) (SEQ ID NO: 171); and Geranyl-Diphosphate Synthase (ERG20WW) (ERG20WW - Enzyme Linker - cTPR6 Spacer - ID Linker - ID 13) (SEQ ID NO: 172).

FIG. 13C contains the amino acid sequences of scaffold-binding engineered enzymes, a soluble CBDA synthase (SEQ ID NO: 173), and a soluble CBCA synthase (SEQ ID NO: 174) encoded by the CAN gene cassette. The scaffold-binding engineered enzymes are Olivetol Synthase (OS) (OS - Enzyme Linker - cTPR6 Spacer - ID Linker -14 כפ) SEQ ID NO: 175); Olivetolic Acid Cyclase (OAC) (OAC - Enzyme Linker cTPR6 Spacer - ID Linker - ID15) (SEQ ID NO: 176); CBGA Synthase (CBGAS Enzyme Linker - cTPR6 Spacer - ID Linker - ID16) (SEQ ID NO: 177); and Acetyl-CoA Carboxylase (ACC) (ACC - Enzyme Linker - cTPR6 Spacer - ID Linker - ID 17) (SEQ ID NO: 178).

FIG. 13D contains the amino acid sequences of the Cannabinoidergic Metabolon

Scaffold (CBSCFLD) - (Myc)3 (SEQ ID NO:179) and the Malonyl-CoAMetabolon

Scaffold (MCASCFLD) - (FLAG)3 (SEQ ID NO: 180).

FIG. 14A contains codon-optimized nucleotide sequences (SEQ ID NOs: 181-187) encoding the enzymes of FIG. 13 A.

FIG. 14B contains the codon-optimized nucleotide sequences (SEQ ID NOs: 188194) encoding the enzymes of FIG. 13B.

FIG. 14C contains the codon-optimized nucleotide sequences (SEQ ID NOs: 195200) encoding the enzymes of FIG. 13C.

FIG. 14D contains the codon-optimized nucleotide sequences (SEQ ID NO:201 and SEQ ID NO:202) encoding the scaffolds of FIG. 13D.

FIG. 15A contains the nucleotide sequence of the HCA gene cassette (SEQ ID NO:203).

FIG. 15B contains the nucleotide sequence of the GPP gene cassette (SEQ ID NO:204).

FIG. 15C contains the nucleotide sequence of the CAN gene cassette (SEQ ID NO:205).

FIG. 15D contains the nucleotide sequence of the SCF gene cassette (SEQ ID NO:206).

FIG. 15E contains the nucleotide sequence of the SOL gene cassette (SEQ ID NO:207).

FIG. 16 is a map of the pCCI-Brick plasmid construct.

FIG. 17 is a map of a pESC-TRP (“vHCA”) vector construct. In this map, the vector contains a TRP gene allowing selection in tryptophan deficient media. Similar vectors also were made in which the TRP gene was replaced with a LEU gene allowing selection in leucine deficient media, a HIS3 gene allowing selection in histidine deficient media, or a URA3 gene allowing selection in uracil deficient media.

FIG. 18 is a graph of the proliferation curves for yCBSCF and yCBSOL cultures. Line plots depicting cell proliferation curves were fitted via nonlinear regression of cell density measurements (OD600nm) recorded in 12-hour intervals over a 48-hour incubation

period for yCBSCF and yCBSOL cultures. Initial cell densities for all cultures were standardized to OD600nm = 0.3. For all measures, n = 3 biological replicates for yCBSCF and yCBSOL cultures. Floating data points depict means with 95% confidence intervals.

Dotted lines represent 95% confidence intervals for regression curve fits.

FIG. 19 shows a comparison of cannabinoid and precursor titers for scaffolded and soluble cannabinoid biosynthesis. Representative mass spectra of target analytes isolated from (A) yCBSOL and (B) yCBSCF cultures incubated for 48 hours in basal culture media. Bar plots depicting (C) Total (aggregate) cannabinoid (CBGA+ CBDA + CBCA + CBG + CBD + CBC) titers, (D) cannabinoid precursor (OVA) titers and summated parent and decarboxylation derivative (CBGA+ CBG; CBDA+ CBD, and CBCA+ CBC) cannabinoid titers, and (E) separated parent (COO(H)) cannabinoid (CBGA, CBDA, and CBCA) and decarboxylation derivative (ACOOH) cannabinoid (CBG; CBD, and CBC) titers for 48-hour yCBSOL (left) and yCBSCF (right) cultures grown in basal culture media. For all measures, n = 3 biological replicates for yCBSCF and yCBSOL cultures. CB, cannabinoid; Cannabigerolic acid, CBGA; cannabigerol, CBG; cannabidiolic acid, CBDA; cannabidiol, CBD; cannabichromenic acid, CBCA; cannabichromene, CBC; olivetolic acid, OVA. Floating asterisks indicate statistically significant (determined by Bonferroni’s multiple comparisons post-hoc test; a = 0.05) between-strain differences for yCBSCF versus yCBSOL cultures. Bar plots depict means with 95% confidence intervals. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

FIG. 20 is a bar plot of the impact of citrate and hexanoate supplementation on scaffolded and soluble cannabinoid biosynthesis. Total cannabinoid (CBGA+ CBDA + CBCA + CBG + CBD + CBC) titers are shown for yCBSOL and yCBSCF cultures incubated for 48 hours in basal, hexanoate (300mg/L)-supplemented, and buffered (pH 6.0) citrate (300mg/L)-supplemented culture media. Floating asterisks indicate statistically significant (determined by Bonferroni’s multiple comparisons post-hoc test; a = 0.05) between-strain differences for yCBSCF versus yCBSOL cultures. Lines with asterisks indicate statistically significant (determined by Bonferroni’s multiple comparisons post-hoc test; a = 0.05) within-strain differences for basal media total cannabinoid titers versus citrate-supplemented media total cannabinoid titers for yCBSCF

cultures. Bar plots depict means with 95% confidence intervals. *p<0.05; **p<0.01;

***p<0.001; ****p<0.0001.

FIG. 21 shows concentration-response parameterization of scaffolded and soluble cannabinoid biosynthesis from citrate. In FIG. 21 A, line plots are shown depicting eightpoint concentration ([citrate]) - response (total cannabinoid titers) curves fitted via asymmetric sigmoidal (five-parameter) logistic regression and in FIG. 2IB, bar graphs are shown depicting concentration-response parameter estimates (CBMax, the estimated maximum total cannabinoid titers and citrate EC50, the estimated citrate concentration yielding half-maximal total cannabinoid titers) for 48-hour yCBscr and yCBsoL cultures incubated for 48 hours in culture media supplemented with 0, 10, 30, 100, 300, 1000, 3000, or 10000 mg/L buffered (pH 6.0) citrate. For all measures, n = 3 biological replicates for yCBscr and yCBsoL cultures. Floating asterisks indicate statistically significant (determined by Bonferroni’s multiple comparisons post-hoc test; a = 0.05) between-strain differences for yCBscr versus yCBsoL cultures. Floating data points and bar plots depict means with 95% confidence intervals. Dotted lines represent 95% confidence intervals for regression curve fits. *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.

DETAILED DESCRIPTION

This document provides methods and materials for producing cannabinoids in host cells or in vitro using a bidirectional, multi-enzymatic scaffold, which can control the localization and stoichiometry of enzymes catalyzing the biosynthesis of cannabinoids and cannabinoid precursors. As described herein, one or more cannabinoids including cannabigerolic acid (CBGA), cannabidiolic acid (CBDA), cannabichromenic acid (CBCA), and tetrahydrocannabinolic acid, can be produced using a bidirectional, multi-enzymatic scaffold and one or more soluble cannabinoid synthesis enzymes, and the conjugate bases, cannabigerolate, cannabidiolate, cannabichromenate, and tetrahydrocannabinolate, respectively, and decarboxylation products, cannabigerol (CBG), cannabidiol (CBD), cannabichromene (CBC), and tetrahydrocannabinol, respectively, of these cannabinoids also can be produced, as can the

tetrahydrocannabinolic acid oxidation product cannabinolic acid and its decararboxylation product cannabinol. The bidirectional, multi-enzymatic scaffold described herein results in significant increases in cannabinoid production in recombinant hosts, including total cannabinoid, CBGA, CBG; CBDA, CBD, CBCA, CBC, and olivetolic acid precursor production, as compared with cannabinoid production in recombinant hosts using the same enzymes that are not bound to a scaffold. As used herein, enzymes that are not bound to a scaffold are referred to as soluble or nonscaffolded. While one particular form of a cannabinoid or other compound may be referenced herein, it is understood that any of its neutral or ionized forms, including any salt forms thereof or decarboxylation derivatives thereof (e.g., produced in the presence of heat and light), are included unless otherwise indicated. It is understood by those skilled in the art that the specific form will depend on factors such as pH and carboxylation status.

In general, enzymes described herein, which can be co-localized on one or more scaffolds and used for producing cannabinoids or cannabinoid precursors, are engineered to contain an interaction domain (ID), which can be separated from the enzyme by an amino acid spacer sequence at the N- or C-terminus of the enzyme. The ID can be composed of two or more scaffold-binding motifs. The engineered enzymes also can include one or more linkers between the enzyme, spacer, and/or ID. The engineered enzymes can bind to a scaffold, which is a polypeptide that contains unique ID-binding domains, i.e., tandem peptide ligands, as shown in FIG. 1A and FIG. IB, such that the enzymes are co-localized to the scaffold. In other words, each enzyme can be engineered to contain a protein-protein interaction domain that is specific for ligand or ligands (binding site) on the scaffold such that the enzyme can be localized to a discrete location along the scaffold via non-covalent interactions. In some cases, the engineered enzymes can be chimeric enzymes. The scaffolded ligands can be separated using amino acid linkers or spacers. See, for example, Horn and Sticht, Frontiers in Bioengineering and Biotechnology, 2015, volume 3, article 191; Whitaker and Dueber, Methods in Enzymology, Chapter 19, “Metabolic Pathway Flux Enhancement by Synthetic Protein

Scaffolding,” Volume 497, 2011, for descriptions of IDs, binding domains, linkers and spacers. IDs also can be referred to as adaptor domains.

Typically, each interaction domain consists of two tandem scaffold-binding motifs that continue/extend from the C-terminus of the engineered enzyme and that can bind to their corresponding scaffolded peptide ligands, which are constructed in tandem along the scaffold. Dual-binding of enzymes to the scaffold ensures fixed spatial orientation, increases binding specificity for each ID-scaffold interaction, and better tethers each enzyme to the scaffold, all of which can improve pathway flux by enabling substrate channeling through each enzymatic step in the scaffolded biosynthetic pathways.

In some embodiments, there are more than two, e.g., three, four, five, six, seven, eight, nine, or ten, or more molecules of each enzyme localized to the scaffold. In addition, the ratio of any given enzyme in a biosynthetic pathway to any other enzyme in the biosynthetic pathway can be varied. For example, the ratio of one engineered enzyme in a pathway to a second engineered enzyme in the same pathway can be varied, e.g., from about 1:5 to about 5:1, e.g., from about 1:5 to about 2:5, from about 2:5 to about 3:5, from about 3:5 to about 5:5, from about 5:5 to about 5:3, from about 5:3 to about 5:2, or from about 5:2 to about 5:1.

The peptide ligands are typically short peptide sequences, ranging in length from 3 to 50 amino acid residues. For example, a peptide ligand can be 3-10, 7-15, 10-20, 1525, 20-30, 25-35, 30-40, 35-45, or 40-50 amino acids in length. There is a database of over 200 different motifs available on the web at elm.eu.org that can be used as described herein. See, for example, Dinkel et aL, Nucleic Acids Res. 2014; 42(Database issue): D259-D266.

An ID can be a peptide sequence ranging in length 3 to 200 amino acid residues. For example, the ID can be 3-10, 7-15, 10-20, 15-25, 20-30, 25-35, 30-40, 35-45, 40-50, 45-55, 50-60, 65-75, 70-80, 85-95, 90-100, 100-110, 105-115, 110-120, 115-125, 120130, 125-135, 130-140, 135-145, 140-150, 135-145, 140-150, 145-155, 150-160, 165175, 170-180, 175-185, 180-190, 185-195, or 190-200 amino acids in length. For example, an ID can be a SH2 domain, a SH3 domain, a PDZ domain, a GTPase binding domain (GBD), a leucine zipper domain, a PTB domain, an FHA domain, a WW domain,

a 14 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosinephenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB1 domain, a SOCS box, an RGS domain, a Toll/IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, a fibronectin receptor domain, a FNDC domain, a SAMD domain, a WBP domain, and/or a SASH domain. See, e.g., U.S. Patent No. 9,856,460 for a list of domains that can be uses as an ID as described herein.

For example, an ID can be a “Src homology2” (SH2) or a “Src homology3” (SH3) domain. SH2 domains are highly conserved structures of approximately 100 amino acid residues that comprise two a-helices and seven β-strands. The SH2 domain can have a promiscuous or strict specificity for a 3-5 amino acid motif flanking a phosphorylated tyrosine. See, Horn and Sticht, 2015, supra. For example, a SH2 domain that can be used as an ID as described herein can be residues 5-122 of a mouse CtlO regulator of kinase adaptor (Crk) protein having GenBank Accession No. AAH31149.

SH3 domains are small modules of approximately 60 residues that bind prolinerich ligands, which bind to the domain surface at three shallow grooves formed by conserved aromatic residues and exhibit two different binding orientations. See, Horn and Sticht, 2015, supra. In some embodiments, the proline-rich ligand can have a core PXXP motif flanked by a positively charged residue. Class IPZP domains recognize ligands conforming to the consensus +XXPXXP (where + is either Arg or Lys), while Class II domains recognize PXXPX+ motifs and bind to ligands in the opposite orientation. See, Teyra, et al., FEBSLett., 2012 586(17):2631-7. Individual SH3 domains do not measurably interact with other SH3 domain family ligands within an organism, minimizing cross-talk and increasing the number of domain/ligand pairs available for simultaneous use. See, Whitaker and Dueber, 2011, supra. For example, a SH3 domain that can be used as an ID as described herein can be residues 134-190 of a mouse Crk

protein having GenBank Accession No. AAH31149 and its peptide ligand can be

PPPALPPKRRR (SEQ ID NO: 1).

For example, an ID can be a PDZ (PSD-95/Discs-large/ZOl) domain. PDZ domains are approximately 100 amino acid residues in length and target specific motifs at the C-terminus of the binding partner. The peptide ligand adopts a β-strand and extends an existing β-sheet within the PDZ domain upon binding. At least four different classes of ligands are known for PDZ domains exhibiting a distinct binding specificity. See, Horn and Sticht, 2015, supra. For example, grouped PDZ domains into two main specificity classes based on distinct ligand signatures: Class I PDZ domains recognize a (X[T/S]X(|)COOH) motif, Class II PDZ domains recognize a (XφXφCOOH) motif, and Class III PDZ domains recognize a X[ED]XφCOOH motif, where X is any residue and φ is a hydrophobic amino acid. See, Teyra, etaL, 2012, supra. PDZ and SH3 domains are found throughout eukaryotic and eubacterial genomes. For example, a PDZ domain that can be used as an ID as described herein can be residues 77-171 of a mouse a-syntrophin protein having GenBank Accession No. EDL06069 and the peptide ligand can be GVKESLV (SEQ ID NO:208).

For example, an ID can be a GBD domain from a protein such as the WiskottAldrich syndrome-like protein (N-WASP). Isolated GBD domains do not adopt a single, discrete structure under physiological conditions but rather exhibit multiple, loosely packed conformations in solution. The corresponding peptide ligand has been deduced from the autoinhibited form of the GBD. See, Hom and Sticht, 2015, supra. For example, a GBD domain that can be used as an ID described herein can include residues 196 to 274 of a rat N-WASP protein having GenBank Accession No. BAA21534, and its peptide ligand, which can be LVGALMHVMQKRSRAIHSSDEGEDQAGDEDED (SEQ ID NO:2), can be used as a peptide ligand as described herein.

For example, an ID can have a leucine zipper or synthetic coiled-coil domain. A leucine zipper domain can include multiple interspersed leucine residues approximately seven amino acid residues apart. Havranek, and Harbury ((2003), Nat. Struct. Biol. 10, 45-52) identified new pairs of homodimers or heterodimers by altering residues between leucine zipper pairs based on computational prediction. Reinke, et al. ((2010). J. Am.

Chem. Soc. 132, 6025-6031) identified three pairs of synthetic coiled coils that do not exhibit measurable self-association. See, Whitaker and Dueber, 2011, supra. One example of an ID that can be used as described herein can be ITIRAAFLEKENTALRTEIAELEKEVGRCENIVSKYETRYGPL (SEQ ID NO:3), and its peptide ligand for use as described herein can be LEIRAAFLEKENTALRTRAAELRKRVGRCRNIVSKYETRYGPL (SEQ ID NO:4).

For example, an ID can be a dockerin polypeptide, which can localize to a specific cohesion polypeptide on a scaffold described herein. Cohesion-dockerin pairs are particularly useful for ex vivo applications as binding is calcium dependent. See, Whitaker and Dueber, 2011, supra.

Combinations of IDs that have high affinity for their peptide ligands and high specificity, i.e., minimal cross-reactivity, can be used as described herein to allow for binding of multiple, different enzymes to a scaffold provided herein. For example, at least three different enzymes can be localized on a scaffold. In some embodiments, at least four different enzymes can be localized on a scaffold. In some embodiments, at least five different enzymes can be localized on a scaffold. In some embodiments, at least six different enzymes can be localized on a scaffold. In some embodiments, at least seven different enzymes can be localized on a scaffold. In some embodiments, at least eight different enzymes can be localized on a scaffold. In some embodiments, at least nine different enzymes can be localized on a scaffold. In some embodiments, at least ten different enzymes can be localized on a scaffold. In some embodiments, at least eleven different enzymes can be localized on a scaffold. In some embodiments, at least twelve different enzymes can be localized on a scaffold. In some embodiments, at least fifteen different enzymes can be localized on a scaffold. In some embodiments, at least seventeen different enzymes can be localized on a scaffold. In some embodiments, at least eighteen different enzymes can be localized on a scaffold. In some embodiments, at least twenty different enzymes can be localized on a scaffold. In some embodiments, at least twenty-one different enzymes can be localized on a scaffold.

Table 1 provide exemplary combinations of heterologous IDs, i.e., IDs that are different from each other, that can be used in seventeen different engineered enzymes and

Table 2 provides the corresponding exemplary combinations of peptide ligands that can be used to localize the seventeen different enzymes to one or more scaffolds. In the embodiments shown in Tables 1 and 2, each ID is composed of two tandem peptide motifs as are the corresponding peptide ligands, which interact with the tandem peptide motifs. It will be appreciated that any one of the enzymes listed in Tables 1 and 2 can be used in combination with any of the listed combinations of IDs and corresponding peptide ligands.

<td> ID Motif #2 Amino Acid Sequence</td><td> SYYHHHHHHLESTSLYKKAGSGS ARNAYLRKKIARLKKDNLQLERD EQNLEKIIANLRDEIARLENEVASH EQ (SEQ ID NO:6)</td><td> SYYHHHHHHLESTSLYKKAGSGS QI&lt;VAELI&lt;NRVAVI&lt;LNRNEQLI&lt;NI&lt; VEELKNRNAYLKNELATLENEVA RLENDVAE (SEQ ID NO :8)</td><td> MAVSESQLKKMVSKYKYRDLTVR ETVNVITLYKDLKPVLDSYVFNDG S SRELMNLTGTIP VP YRGNT YNIPI CLWLLDTYPYNPPICFVKPTSSMTI I&lt;TGI&lt;HVDANGI&lt;IYLPYLHEWI&lt;HP QSDLLGLIQVMIVVFGDEPPVFSRP (SEQ ID NO: 10)</td><td> NTNMSVPTDGAVTTSQIPASEQET LVRPKPLLLKLLKSVGAQKDTYT MKEVLFYLGQYIMTKRLYDEKQQ HIVYCSNDLLGDLFGVPSFSVKEH RKIYTMIYRNLVV (SEQ ID NO: 12)</td>

<td> ID Motif #2</td><td> SYNZIP2</td><td> SYNZIP4</td><td> UEV</td><td> MDM2</td>

<td> ID Motif #1 Amino Acid Sequence</td><td> SYYHHHHHHLESTSLYKKAGSG SNLVAQLENEVASLENENETLK KKNLHKKDLIAYLEKEIANLRK KIEE ((SEQ ID NO:5))</td><td> SYYHHHHHHLESTSLYKKAGSG SNEVTTLENDAAFIENENAYLE I&lt;EIARLRI&lt;EI&lt;AALRNRLAHI&lt;I&lt; (SEQ ID NO:7)</td><td> ENLYFQGENLYFQGDSSESCWN CGRKASETCSGCNTARYCGSFC QHKDWEKHHHICGQTLQAQQ (SEQ ID NO:9)</td><td> GPLGSPLTASMLASAPPQEQKQ MLGERLFPLIQAMHPTLAGKITG MLLEIDNSELLHMLESPESLRSK VDEAVAVLQAHQAKEAAQKA (SEQ ID NO: 11)</td>

<td> ID Motif #1</td><td> SYNZIP1</td><td> SYNZIP3</td><td> MYND</td><td> PABP</td>

<td> S</td><td></td><td> eq</td><td></td><td></td>

<td> Enzyme</td><td> ATP Citrate Lyase</td><td> Acetyl-CoA Acetyltransferase (atoB)</td><td> 3-hydroxybutyrylC0A Dehydrogenase</td><td> Enoyl-CoA Hydratase</td>

<td> SYYHHHHHHLESTSLYKKAGSGS KRIAYLRKKIAALKKDNANLEKDI ANLENEIERLIKEIKTLENEVASHE Q (SEQ ID NO :14)</td><td> ESDSVEFNNAISYVNKIKTRFLDHP EIYRSFLEILHTYQKEQLHTKGRPF RGMSEEEVFTEVANLFRGQEDLLS EFGQFLPEAKR (SEQ ID NO: 16)</td><td> GAMGPLPPGWEKRTDSNGRVYFV NHNTRITQWEDPRS (SEQ ID NO: 18)</td><td> SYYHHHHHHLESTSLYKKAGSGS QKVESLKQKIEELKQRKAQLKNDI ANLEKEIAYAET (SEQ ID NO :20)</td><td> GAMGSMAEAEGESLESWLNKATN PSNRQEDWEYIIGFCDQINKELEGP QIAVRLLAHKIQSPQEWEALQALT VLEACMKNCGRRFHNEVGKFRFL NELIKVVSPKYLGDRVSEKVKTKV IELLYSWTMALPEEAKIKDAYHML KRQGIVQSDPPIPVDRTLIPSPPPRP KN (SEQ ID NO:22)</td><td> SYYHHHHHHLESTSLYKKAGSGSF ENVTHEFILATLENENAKLRRLEA KLERELARLRNEVAWL (SEQ ID</td>

<td> SYNZIP22</td><td> PAH</td><td> WW1B</td><td> SYNZIP9</td><td> VHS2</td><td> SYNZIP15</td>

<td> SYYHHHHHHLESTSLYKKAGSG SNLLATLRSTAAVLENENHVLE KEKEKLRKEKEQLLNKLEAYK (SEQ ID NO: 13)</td><td> DVMWEYKWENTGDAELYGPFT SAQMQTWVSEGYFPDGVYCRK LDPPGGQFYNSKRIDFDLYT (SEQ ID NO: 15)</td><td> LGPLPPGWEVRSTVSGRIYFVD HNNRTTQFTDPRLH (SEQ ID NO: 17)</td><td> SYYHHHHHHLESTSLYKKAGSE FFRRERNKMAAAKCRNRRRELT DTLQAETDQLEDEKSALQTEIA NLLKEKEKLEFILAAHRPACKIP DDLGFPEEMSLE (SEQ ID NO: 19)</td><td> MEPAMEPETLEARINRATNPLN KELDWASINGFCEQLNEDFEGP PL ATRLL AHKIQ SPQEWE AIQ AL TVLETCMKSCGKRFHDEVGKFR FLNELIKVVSPKYLGSRTSEKVK NKILELLYSWTVGLPEEVKIAEA YQMLKKQGIVKS (SEQ ID NO:21)</td><td> SYYHHHHHHLESTSLYKKAGSG SQKVEELKNKIAELENRNAVKK NRVAHLKQEIAYLKDELAAHEF</td>

<td> SYNZIP1 0</td><td> GYF</td><td> WW1A</td><td> FOS</td><td> VHS1</td><td> SYNZIP1 3</td>

<td></td><td> SO</td><td></td><td> 00</td><td> OS</td><td> 10</td>

<td> Trans-Enoyl-CoA Reductase</td><td> Beta-ketothiolase (bktB)</td><td> HMG-C0A Synthase</td><td> HMG-C0A Reductase</td><td> Mevalonate Kinase</td><td> Phosphomevalonate Kinase</td>

<td> NO:24)</td><td> ASIKLQSSDGEIFEVDVEIAKQSVTI KTMLEDLGMDDEGDDDPVPLPNV NAAILKKVIQWCTHHKDDPPPPED DENKEKRTDDIPVWDQEFLKVDQ GTLFELILAANYLDIKGLLDVTCKT VANMIKGKTPEEIRKTFNIKNDFTE EEEAQVRKENQWC (SEQ ID NO:26)</td><td> SYYHHHHHHLESTSLYKKAGSGS QKVAQLKNRVAYKLKENAKLENI VARLENDNANLEKDIANLEKDIAN LERDVAR (SEQ ID NO:28)</td><td> S SGAIIYTVELKRYGGPLGITISGTE EPFDPIIIS SLTKGGLAERTGAIHIG DRIL AINS S SLKGKPL SEAIHLLQM AGETVTLKIKKQTDAQPASS (SEQ IDNO:30)</td><td> GSHPWFFGKIPRAKAEEMLSKQRH DGAFLIRESESAPGDFSLSVKFGND VQHFKVLRDGAGKYFLWVVKFNS LNELVDYHRSTSVSRNQQIFLRDIE QVPQQPT (SEQ ID NO:32)</td>

<td></td><td> SKP1</td><td> SYNZIP6</td><td> PDZ2</td><td> SH2B</td>

<td> E (SEQ ID NO:23)</td><td> AMADLEQKVLEMEASTYDGVFI WKISDFPRKRQEAVAGRIP AIF S PAFYTSRYGYKMCLRIYLNGDG TGRGTHLSLFFVVMKGPNDALL RWPFNQKVTLMLLDQNNREHV IDAFRPDVTSSSFQRPVNDMNIA SGCPLFCPVSKMEAKNSYVRDD AIFIKAIVDLTGL (SEQ ID NO:25)</td><td> SYYHHHHHHLESTSLYKKAGSG SNTVKELKNYIQELEERNAELK NLKEHLKFAKAELEFELAAHKF E (SEQ ID NO:27)</td><td> LCTMKKGPSGYGFNLHSDKSKP GQFIRSVDPDSPAEASGLRAQDR IVEVNGVCMEGKQHGDVVSAIR AGGDETKLLVVDRE (SEQ ID NO:29)</td><td> GNNLETYEWYNI&lt;SISRDI&lt;AEI&lt;L LLDTGKEGAFMVRDSRTPGTYT VSVFTKAIISENPCIKHYHIKETN DSPKRYYVAEKYVFDSIPLLIQY HQYNGGGLVTRLRYPVCG (SEQ IDNO:31)</td>

<td></td><td> MATH</td><td> SYNZIP5</td><td> PDZ1</td><td> SH2A</td>

<td></td><td> 11</td><td> 12</td><td> 13</td><td> 14</td>

<td></td><td> Diphosphomevalonat e Decarboxylase</td><td> Isopentenyl- Diphosphate Delta- Isomerase</td><td> Geranyl-Diphosphate Synthase</td><td> Olivetol Synthase</td>

<td> GSHMGSQFWVTSQKTEASERCGL QGSYILRVEAEKLTLLTLGAQSQIL EPLLF WP YTLLRRYGRDKVMF SFE AGRRCPSGPGTFTFQTSQGNDIFQ AVEAAIQQQKAQGKVGQAQDILR LEHHHHHH (SEQ ID ΝΟ:210)</td><td> LIKHMRAEALFDFTGNSKLELNFK AGDVIFLLSRINKDWLEGTVRGAT GIFPLSFVKILK (SEQ ID NO:35)</td><td> GAMATPGSENVLPREPLIATAVKF LQNSRVRQSPLATRRAFLKKKGLT DEEIDMAFQQSGTAADEPSSLW (SEQ ID NO:37)</td>

<td> eq η Η</td><td> SH3B</td><td> PEX</td>

<td> GQDRSEATLIKRFKGEGVRYKA KLIGIDE VS AARGDKLCQD SMM KLKGVVAGARSKGEHKQKIFLT ISFGGIKIFDEKTGALQHHHAVH EISYIAKDITDHRAFGYVCGKEG NHRFVAIKTAQAAEPVILDLRDL FQLIYELKQREELEKKA (SEQ ID NO:33)</td><td> AEYVRALFDFNGNDEEDLPFKK GDILRIRDKPEEQWWNAEDSEG KRGMIPVPYVEKY (SEQ ID NO:34)</td><td> GSHMRLGAQSIQPTANLDRTDD LVYLNVMELVRAVLELKNELA QLPPEGYVVVVKNVGLTLRKLI GSVDDLLPSLPSSSRTEIEGTQKL LNKDLAELINKMRLAQQNAVTS LSEECKRQMLTASHTLAVDAKN LLDAVDQAKVLANLAHPPAE (SEQIDNO:36)</td>

<td> co H Olh</td><td> SH3A</td><td> FAT</td>

<td> in</td><td></td><td> I—</td>

<td> Olivetolic Acid Cyclase</td><td> CBGA Synthase</td><td> Acetyl-CoA Carboxylase</td>

eq

&amp;#1501;&amp;#1470;

=

&amp;#1501;&amp;#1470; =

Η

<td> ID Motif #2 Scaffolded Ligand Amino Acid Sequence</td><td> SYYHHHHHHLESTSLYKKAGSGS NLVAQLENEVASLENENETLKKK NLHI&lt;I&lt;DLIAYLEI&lt;EIANLRI&lt;I&lt;IEE (SEQ ID NO:5)</td>

<td> ID Motif #2</td><td> SYNZIP2</td>

<td> ID Motif #1 Scaffolded Ligand Amino Acid Sequence</td><td> SYYHHHHHHLESTSLYKKAGS GSARNAYLRI&lt;I&lt;IARLI&lt;I&lt;DNLQ LERDEQNLEKIIANLRDEIARLE NEVASHEQ (SEQ ID NO :6)</td>

<td> ID Motif #1</td><td> SYNZIP1</td>

<td></td><td></td>

<td> Enzyme</td><td> ATP Citrate Lyase</td>

<td> SYYHHHHHHLESTSLYKKAGSGS NEVTTLENDAAFIENENAYLEKEI ARLRI&lt;EI&lt;AALRNRLAHI&lt;I&lt; (SEQ ID NO :7)</td><td> NFLQSRPEPTAPPEESFRSG (SEQ IDNO:39)</td><td> PDGGTTFEHLWSSLEPDSTY (SEQ IDNO:41)</td><td> SYYHHHHHHLESTSLYKKAGSGS NLLATLRSTAAVLENENHVLEKEK EKLRKEKEQLLNKLEAYK (SEQ ID NO:13)</td><td> ELNSLLILLEAAEYLERRDR (SEQ IDNO:43)</td><td> ERESNEEPPPPYEDPYWGNG (SEQ IDNO:45)</td><td> SYYHHHHHHLESTSLYKKAGSEFF RRERNKMAAAKCRNRRRELTDTL QAETDQLEDEKSALQTEIANLLKE KEKLEFILAAHRPACKIPDDLGFPE EMSLE (SEQ ID NO: 19)</td><td> AAATPISTFHDDSDEDLLHV (SEQ IDNO:47)</td>

<td> -r N Z tn</td><td> UEV</td><td> MDM2</td><td> SYNZIP22</td><td> PAH</td><td> WW1B</td><td> SYNZIP9</td><td> VHS2</td>

<td> SYYHHHHHHLESTSLYKKAGS GSQKVAELKNRVAVKLNRNEQ LKNKVEELKNRNAYLKNELAT LENEVARLENDVAE (SEQ ID NO :8)</td><td> RPPTISNPPPLISSAKHPSV (SEQ IDNO:38)</td><td> SKGTGLNPNAKVWQEIAPGN (SEQ ID NO:40)</td><td> SYYHHHHHHLESTSLYKKAGS GSKRIAYLRKKIAALKKDNAN LEKDIANLENEIERLIKEIKTLE NEVASHEQ (SEQ ID NO: 14)</td><td> PATSQHPPPPPGHRSQAPSH (SEQ ID NO:42)</td><td> FQMPADTPPPAYLPPEDPMT (SEQ ID NO:44)</td><td> SYYHHHHHHLESTSLYKKAGS GSQKVESLKQKIEELKQRKAQL KNDIANLEKEIAYAET (SEQ ID NO:20)</td><td> VSSTKLVSFHDDSDEDLLHI (SEQ ID NO:46)</td>

<td> N Z tn</td><td> MYND</td><td> PABP</td><td> SYNZIP1 0</td><td> GYF</td><td> WW1A</td><td> FOS</td><td> VHS1</td>

<td> eq</td><td></td><td></td><td></td><td> SO</td><td></td><td> 00</td><td> OS</td>

<td> Acetyl-CoA Acetyltransferase (atoB)</td><td> 3-hydroxybutyrylC0A Dehydrogenase</td><td> Enoyl-CoA Hydratase</td><td> Trans-Enoyl-CoA Reductase</td><td> Beta-Ketothiolase (bktB)</td><td> HMG-C0A Synthase</td><td> HMG-C0A Reductase</td><td> Mevalonate Kinase</td>

<td> SYYHHHHHHLESTSLYKKAGSGS QI&lt;VEELI&lt;NI&lt;IAELENRNAVI&lt;I&lt;NR VAHLKQEIAYLKDELAAHEFE (SEQ ID NO:23)</td><td> GSPNAGSVEQTPKKPGLRRR (SEQ IDNO:49)</td><td> SYYHHHHHHLESTSLYKKAGSGS NTVKELKNYIQELEERNAELKNLK EHLKFAKAELEFELAAHKFE (SEQ IDNO:27)</td><td> DGNVSGTQRLDSATVRTYSC (SEQ IDNO:51)</td><td> RELFDDPSYVNVQNLDKARQ (SEQ IDNO:53)</td><td> RSLPSTWIENKLYGMSDPNW (SEQ IDNO:55)</td><td> TQRSKPQPAVPPRPSADLIL (SEQ IDNO:57)</td><td> DLALSENWAQEFLAAGDAVD (SEQIDNO:59)</td>

<td> £ N Z</td><td> SKP1</td><td> SYNZIP6</td><td> PDZ2</td><td> SH2B</td><td> PTB2</td><td> SH3B</td><td> PEX</td>

<td> SYYHHHHHHLESTSLYKKAGS GSFENVTHEFILATLENENAKL RRLEAKLERELARLRNEVAWL (SEQ ID NO:24)</td><td> HDDSLPHPQQATDDSGHESD (SEQ ID NO:48)</td><td> SYYHHHHHHLESTSLYKKAGS GSQKVAQLKNRVAYKLKENA KLENIVARLENDNANLEKDIAN LEKDIANLERDVAR (SEQ ID NO:28)</td><td> TDEEREETEEEVYLLNSTTL (SEQIDNO:50)</td><td> ALVDDAADYEPPPSNNEEAL (SEQ ID NO:52)</td><td> KNTKSMNFDNPVYRKTTEEE (SEQIDNO:54)</td><td> VVDNSPPPALPPKKRQSAPS (SEQIDNO:56)</td><td> SATRELDELMASLSDFKIQG (SEQ ID NO:58)</td>

<td> £ S י cn</td><td> MATH</td><td> SYNZIP5</td><td> PDZ1</td><td> SH2A</td><td> PTB1</td><td> SH3A</td><td> FAT</td>

<td> o</td><td></td><td> eq</td><td> 2</td><td></td><td> in</td><td></td><td> I—</td>

<td> Phosphomevalonate Kinase</td><td> Diphosphomevalonate Decarboxylase</td><td> Isopentenyl- Diphosphate Delta- Isomerase</td><td> Geranyl-Diphosphate Synthase</td><td> Olivetol Synthase</td><td> Olivetolic Acid Cyclase</td><td> CBGA Synthase</td><td> Acetyl-CoA Carboxylase</td>

The spacers or linkers connecting an enzyme and ID, as well as a binding domain on a scaffold, can be peptide sequences ranging in length from 6 to 250 amino acid residues. The term “spacer” typically refers to a longer and more structurally-rigid peptide sequence and the term “linker” typically refers to a shorter and more structurallyflexible peptide sequence. In embodiments in which both terms are used, linker typically refers to a sequence that is about 3 to about 50 amino acids in length and spacer typically refers to a sequence that is longer (e.g., about 36 to about 250 amino acids in length). For example, a linker can be 6-15, 10-20, 15-25, 20-30, 25-35, 30-40, 35-45, or 40-50 amino acids in length. A spacer can be, for example, 36-40, 40-50, 45-55, 50-60, 55-65, 60-70, 65-75, 70-80, 75-85, 90-100, 95-105, 100-110, 105-115, 110-120, 115-125, 120-130, 125-135, 130-140, 135-145, 140-150, 145-155, 150-160, 165-175, 170-180, 175-185, 180-190, 185-195, 190-200, 195-205, 200-210, 205-215, 210-220, 215-225, 220-230, 225-235, 230-240, 235-245, or 240-250 amino acids in length. See, for example, Chen, et aL, Adv DrugDeliv Rev. 2013 65(10): 1357-1369. In either case, the linker/spacer can be a series of small and/or hydrophilic and/or other amino acid residues that can adapt flexible and/or rigid structures. For example, the linker can be a series of glycine residues, a series of alanine residues, a series of serine residues, or a series of alternating glycine and serine (or threonine) residues such as (G-S)8 (SEQ ID NO:60), (G-S)io (SEQ ID NO:61), or (G-S)15 (SEQ ID NO:62), or contain mainly glycine residues such as (GGGGS)3 (SEQ ID NO:63) or (GGGGS)4 (SEQ ID NO:64), or contain any other series of canonical or non-canonical amino acid residues or combinations thereof. In some embodiments, a linker can include glutamic acid, alanine, and lysine residues such as (EAAAK)2 (SEQ ID NO:65), (EAAAK)3 (SEQ ID NO:66), or (EAAAK)4 (SEQ ID NO:67). See, Horn and Sticht, 2015, supra. In some embodiments, a linker can be a combination of glycine, alanine, proline and methionine residues, such as AAAGGM (SEQ ID NO:68), AAAGGMPP AAAGGM (SEQ ID NO:69), AAAGGM (SEQ ID NO:70), orPPAAAGGMM (SEQ IDNO:71). See, e.g., U.S. Patent No. 9,856,460.

Based on amino acid composition, linkers or spacers can be either structured or intrinsically unstructured. For example, in some embodiments, a spacer can have a sequence that adopts a more structurally-rigid a-helical conformation and a linker can

have a GS-rich peptide sequence that is more structurally-flexible. For example, in some embodiments, a linker can include flexible GS-rich sequences flanking one or more rigid a-helical moieties, e.g., GS-rich sequences flanking duplicate, triplicate, or quadruplicate a-helical moieties. For example, in some embodiments, a linker or spacer can have the sequence GSAGSAAGSGEF (SEQ ID NO:72), KLSGGGGSGGGGSGGGGS (SEQ ID NO:73), GSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGS (SEQ ID NO:74), GSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEF (SEQ ID NO:75), or GSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGS (SEQ ID NO:76).

In some embodiments, the ligands on the scaffold can be separated by linkers that are 20-50 amino acid residues in length (e.g., 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33. 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acid residues in length). In some embodiments, the IDs engineered at the C-terminus or Nterminus of each scaffolded enzyme can contain a linker (e.g., a flexible linker) of 15 to 30 (e.g., 20) amino acid residues in length flanking a spacer of 15 to 50 (e.g. 36) amino acid residues. In some embodiments, the ID can be separated from the enzyme by a spacer sequence such as the cTPR6 spacer, which includes sextuplicate rigid a-helical moieties and can have the sequence: AEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDY QKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNL QAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGD YQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNN RSRSA (SEQ ID NO:77).

In some embodiments, the engineered enzyme can be of a formula: enzyme linker! - spacer - linker2 - motifi - linkers - motif2, where linkers 1, 2, and 3 can be the same or different, and motif 1 and motif 2 can be the same or different. In some embodiments, linker 1 can be referred to as the enzyme linker, i.e., it connects the enzyme to the spacer such as cTPR6 spacer, and can include flexible GS-rich moieties flanking a rigid a-helical moiety such as KLSGGGGSGGGGSGGGGS (SEQ ID NO:73). In some embodiments, linker 2 can be referred to as the ID linker and can include, for

example, flexible GS-rich moieties flanking a rigid a-helical moiety such as GGGGSGGGGSGGGGAS (SEQ ID NO:78). In some embodiments, linker 3 can be referred to as the motif linker and can include flexible GS-rich moieties flanking a rigid a-helical moiety such as GSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGS (SEQ ID NO:74). Table 1 provides non-limiting examples of motifs 1 and motifs 2, which are used together to form heterologous IDs. FIG. 3 contains a schematic of an exemplary engineered enzyme of this formula complexed with a scaffold. FIG. 6B and FIGs. 13 A-C contain the amino acid sequence of an ATP citrate lyase, atoB, a 3-hydroxybutyryl-CoA dehydrogenase, an enoyl-CoA hydratase, a trans-enoyl-CoA reductase, a betoketothiolase (bktB), an HMG-C0A synthase, a truncated HMG-C0A reductase, a mevalonate kinase, a phosphomevalonate kinase, a diphosphomevalonate decarboxylase, an isopentenyl-diphosphate delta isomerase, a geranyl-diphosphate synthase (ERG20ww), an olivetol synthase, an olivetolic acid cyclase, a CBGA synthase, and an acetyl-CoA carboxylase according to this formula. In some embodiments, linkers 1 and 2 can be (G4S)3, the spacer can be the cTPR6 sequence, and linker 3 can be (GS)8.

In some embodiments, a scaffold can be of a formula: N-terminus-[Ligand #1 linker - Ligand #2 -Spacer]n -(optionally-tagged) C-terminus, where n is the number of interaction domains. The linker can be referred to as a scaffolded ligand linker and can be used to connect and separate paired motif-binding ligands that recruit/localize each enzyme to its scaffold-binding site. Such a linker can include flexible GS-rich moieties flanking a rigid a-helical moiety and have a sequence such as GSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEF (SEQ ID NO:75). The spacer can be referred to as a scaffolded ID-binding site spacer and can be used to connect and separate the scaffold-binding sites (composed of the paired motif binding ligands) for each enzyme. Such a spacer can include flexible GS-rich moieties flanking a rigid a-helical moiety and have a sequence such as GSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGS (SEQ ID NO:76). The N-terminus can include a flexible GS-rich sequence to help stabilize and solubilize the scaffold. For example, the N-terminus can have the sequence

GSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEF (SEQ ID NO:79). The Cterminus can include a flexible GS rich sequence flanking a rigid a-helical moiety to stabilize and solubilize the scaffold and can be optionally tagged (e.g., with a MYC tag, a FLAG tag, or other tag described below) to ease purification or detection of the scaffold. For example, a C-terminal sequence with a triplicate MYC tag can have the sequence GSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSEQK LISEEDLEQKLISEEDLEQKLISEEDLGSAGSAAGSGEFGSAGSAAGSGEFGSAGS AAGSGEF (SEQ ID NO:80). For example, a C-terminal sequence with a triplicate FLAG tag can have the sequence GSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSDYK DDDDKDYKDDDDKDYKDDDDKGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAA GSGEF (SEQ ID NO:81). FIG. 6C and FIG. 13D each contain an example of a scaffold polypeptide of this formula that contains the peptide ligands corresponding to IDS 1-16 as shown in Table 2, and a triplicate MYC tag on the C-terminus. For example, FIG. 13D contains an example of a scaffold polypeptide (see SCF gene cassette of FIG. 2B) containing a triplicate MYC tag. FIG. 6D and FIG. 13D each contain an example of a scaffold polypeptide that contains the peptide ligands corresponding to IDs 1 and 17 as shown in Table 2 and a triplicate FLAG tag on the C-terminus. Accordingly, the amino acid sequence of a scaffold can depend on the sequence of the peptide ligands that can bind to the selected ID motif of the enzymes.

In some embodiments, any one of the enzymes can be engineered to include an Nterminal or C-terminal linker motif that allows covalent (isopeptide) bonding to the scaffold. See, for example, the SpyTag and SpyCatcher system described by Zakeri, et al., Proc. Natl. Acad. Sci., 2012 109 (12) E690-E697.

In some embodiments involving multi-enzymatic scaffolds described herein, the first engineered enzyme of a biosynthetic pathway can produce a first product that can be a substrate for the second engineered enzyme of the biosynthetic pathway, the second engineered enzyme of the biosynthetic pathway can produce a second product that can be a substrate for the third engineered enzyme of the biosynthetic pathway, and so forth. In some cases, the second engineered enzyme can be immobilized on the scaffold such that

it is positioned adjacent to or very close to the first engineered enzyme. The third engineered enzyme can be immobilized on the scaffold such that it is positioned adjacent or very close the second engineered enzyme. In this way, the effective concentration of the first product can be high, and the second engineered enzyme can act efficiently on the first product, the third engineered enzyme can act efficiently on the second product, and so forth.

As shown in FIGs. 1A and IB, one example of a multi-enzymatic scaffold contains enzymes of the hexanoyl-CoA pathway on the N-terminus of the scaffold, enzymes of the mevalonate pathway on the C-terminus of the scaffold, and enzymes of the upper cannabinoid pathway in between. Within any of the pathways, the enzymes can be from a single source, i.e., from one species or genera, or can be from multiple sources, i.e., different species or genera. Nucleic acids encoding the enzymes described herein have been identified from various organisms and are readily available in publicly available databases such as GenBank or EMBL (see below).

A fully-assembled multi-enzymatic scaffold provided herein can adopt stoichiometry and a spatial arrangement that can help maximize pathway flux and minimize accumulation of pathway intermediates and by-products. Such scaffolds can facilitate substrate channeling both within and between cannabinoid and cannabinoid precursor pathways. Specifically, this scaffolding system can facilitate unidirectional flux through each of the primary cannabinoid precursor pathways, and converging near the midpoint of the scaffold. The hexanoyl-CoA/olivetolic acid (OVA) pathway can begin at the N-terminus of the scaffold, and the mevalonate or MEP pathway can begin at the Cterminus of the scaffold. The enzyme catalyzing the rate-limiting/committed step in cannabinoid biosynthesis, a CBGA synthase, can be localized at the intersection of these precursor pathways near the scaffold midpoint.

By this design, the two primary precursors for cannabinoid biosynthesis, hexanoyl-CoA/olivetolic acid and geranyl pyrophosphate, can be bi-directionally delivered to a CBGA synthase at this intersection. The CBGA synthase can catalyze biosynthesis of CBGA, the primary cannabinoid from which all other cannabinoids are

derived. Substrate channeling within and between the scaffolded pathways can accelerate the kinetics of the composite pathway in accordance with the law of mass action.

In the embodiment shown in FIGs. 1A and IB, the N-terminal hexanoyl-CoA pathway can include an ATP citrate lyase (ACL) (also can be referred to as an ATP citrate synthase), an acetyl-CoA acetyltransferase (atoB), two 3-hydroxy-acyl-CoA dehydrogenases (BHBDs), two enoyl-CoA hydratases (ECHs), a beta-ketothiolase (bktB), and two trans enoyl-CoA-reductases (ECRs).

In the hexanoyl-CoA pathway shown in FIGs. 1A and IB, citrate, from cellular metabolism and/or supplemented in the growth medium, can be used as a substrate for ACL-catalyzed acetyl-CoA synthesis. ACL is classified under EC 2.3.3.8. Acetyl-CoA can be used as a substrate for atoB-catalyzed acetoacetyl-CoA synthesis. atoB is classified under EC 2.3.1.9. Acetoacetyl-CoA can serve as the substrate for BHBDcatalyzed 3-hydroxybutanoyl-CoA synthesis. BHBD is classified under EC 1.1.1.157. 3hydroxybutanoyl-CoA can serve as the substrate for ECH-catalyzed trans-but enoylC0A synthesis. ECH is classified under EC 4.2.1.17. Trans-but enoyl-CoA can serve as the substrate for ECR-catalyzed butanoyl-CoA synthesis. ECR is classified under EC 1.3.8.1. Butanoyl-CoA can serve as the substrate for bktB-catalyzed 3-keto-hexanoy 1C0A synthesis. bktB is classified under EC 2.3.1.9. The bktB catalyzing the production of 3-ketohexanoyl C0A from butanoyl-CoA can be the same as, or different from, the atoB used to catalyze the production of acetoacetyl-CoA from acetyl-CoA. 3-ketohexanoylC0A is the substrate for BHBD-catalyzed 3-hydroxyhexanoyl-CoA synthesis. BHBD is classified under EC 1.1.1.157. The BHBD catalyzing the production of 3hydroxyhexanoyl-CoA can be the same as, or different from, the BHBD used to catalyze the production of 3-hydroxybutanoyl-CoA. 3-hydroxyhexanoyl-CoA can be the substrate for ECH-catalyzed trans-hex enoyl-CoA synthesis. ECH is classified under 4.2.1.17. The ECH catalyzing the production of trans-hex enoyl-CoA can be the same as, or different from, the ECH used to catalyze the production of trans-but enoyl-CoA. Transhex enoyl-CoA can be the substrate for ECR-catalyzed hexanoyl-CoA synthesis. ECR is classified under EC 1.3.1.38 or EC 1.3.1.44. The ECR catalyzing the production of

hexanoyl-CoA can be the same as, or different from, the ECR used to catalyze the production of butanoyl-CoA

In some embodiments, a hexanoyl-CoA synthetase (HCS) enzyme can be substituted for the scaffolded enzymes of the hexanoyl-CoA pathway or can be included in a soluble form in addition to the scaffolded enzymes of the hexanoyl-CoA pathway, and in some embodiments, hexanoic acid can be added to the growth media as a substrate for HCS-catalyzed hexanoyl-CoA production. The HCS can be included on the scaffold, N-terminal to the upper cannabinoid pathway in FIGs. 1A and IB, and/or it can be nonscaffolded (soluble).

In the embodiment shown in FIGs. lAand IB, the C-terminal mevalonate pathway can include an ACL, an atoB, a hydroxymethylglutaryl-CoA, an HMG-C0A synthase (HMGS), an HMG-C0A reductase (HMGR), a mevalonate kinase (ERG12), a phosphomevalonate kinase (ERG8), a diphospho mevalonate decarboxylase (MVD1), an isopentyl diphosphate isomerase (IDI1), and a mutant GPP synthase (mGPPS). In the mevalonate pathway shown in FIGs. 1A and IB, citrate from cellular metabolism and/or supplemented in the growth medium, can be used as a substrate for ACL-catalyzed acetyl-CoA synthesis. ACL is classified under EC 2.3.3. Acetyl-CoA can be used as a substrate for bktB-catalyzed acetoacetyl-CoA synthesis. bktB is classified under EC 2.3.1.9. Acetoacetyl-CoA can be the substrate for HMGS-catalyzed HMG-C0A synthesis. HMG-C0A can be the substrate for HMGR catalyzed mevalonate synthesis. HMGR is classified under EC 1.1.1.88 or 1.1.1.34. Mevalonate can be the substrate for mevalonate kinase-catalyzed mevalonate-5 phosphate synthesis. Mevalonate kinase is classified under EC 2.7.1.36. Mevalonate phosphate can be the substrate for phosphomevalonate kinase-catalyzed mevalonate pyrophosphate synthesis. Phosphomevalonate kinase is classified under EC 2.7.4.2. Mevalonate pyrophosphate can be the substrate for diphosphomevalonate decarboxylase-catalyzed isopentyl pyrophosphate synthesis. Diphosphomevalonate decarboxylase is classified under EC 4.1.1.33. Isopentyl pyrophosphate can be the substrate for isopentyl diphosphate isomerase-catalyzed dimethylallyl pyrophosphate synthesis. Isopentyl diphosphate isomerase is classified under EC. 5.3.3.2. Dimethylallyl pyrophosphate can be the substrate for geranyl

pyrophosphate synthase (GPPS)-catalyzed geranyl pyrophosphate synthesis. GPPS is classified under EC 2.5.1.1.

As acetyl-CoA can be the initial substrate for the hexanoyl-CoA, mevalonate/geranyl pyrophosphate, and malonyl-CoA cannabinoid precursor biosynthetic pathways, the inclusion of ACL at both the N-terminus and C-terminus of the multi-enzymatic scaffold in FIGs. 1A and IB can directly couple the scaffolded pathways to cellular metabolism via ACL-catalyzed production of acetyl-CoA from citric acid cycle-derived citrate. The citrate also can be supplemented into the culture medium (e.g., as buffered citrate). In some embodiments, the ACL enzyme is included only at the N-terminus of the scaffold. In some embodiments, the ACL enzyme is included only at the C-terminus of the scaffold. In some embodiments, the ACL enzyme is included in soluble form.

In some embodiments, the 2-C-methylerythritol 4-phosphate (MEP) pathway, which also can produce geranyl pyrophosphate, can be substituted for the scaffolded mevalonate pathway at the C-terminus of the scaffold or can be included in a soluble form in addition to the scaffolded mevalonate pathway. For example, as shown in FIG. 5, the C-terminus of the scaffold can include a 1-deoxy-D-xylulose phosphate (DOXP) synthase, a DOXP reductoisomerase, a MEP cytidyl transferase, a 4-diphosphocytidyl-2C-methylerythritol (CDPME) kinase, a 2-C-methyl-D-erythritol 2,4-cyclodiphosphate (MECDP) synthase, a 4-hydroxy methyl-but enyl pyrophosphate (HMBPP) synthase, a HMBPP reductase, and a GPPS. Pyruvate and glyceraldehyde phosphate (G3P) can be used as substrates for DOXP-synthase-catalyzed DOXP synthesis. DOXP is classified under EC 2.2.1.7. DOXP can be the substrate for DOXP reductoisomerase (DXR)catalyzed MEP synthesis. DXR is classified under EC 1.1.1.267. MEP can be the substrate for 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (ISPD)-catalyzed 4-diphosphocytidyl C-methylerythritol (CDP-ME) synthesis. ISPD is classified under EC 2.7.7.60. CDP-ME can be the substrate for 4-diphosphocytidyl C-methyl-Derythritol kinase (ISPE)-catalyzed 4-diphosphocytidyl C-methyl-D-erythritol 2phosphate (CDP-MEP) synthesis. ISPE is classified under EC 2.7.1.148. CDP-MEP can be the substrate for 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (ISPF)34

catalyzed 2-C-methyl-D-erythritol 2,4-cyclodiphosphate (cMEPP) synthesis. ISPF is classified under EC 4.6.1.12. cMEPP can be the substrate for HMB-PP synthase (ISPG)catalyzed (E) Hydroxy methyl-but enyl pyrophosphate (HMBPP) synthesis. ISPG is classified under EC 1.17.7.1. HMBPP can be the substrate for 4-hydroxy methylbut2-enyl diphosphate reductase (ISPH)-catalyzed isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) synthesis. ISPH is classified under EC 1.17.1.2. IPP and DMAPP can be substrates for GPPS-catalyzed geranyl pyrophosphate synthesis. GPPS is classified under EC 2.5.1.1.

In some embodiments, the mevalonate pathway can be substituted for the scaffolded MEP pathway at the C-terminus of the scaffold or can be included in a soluble form in addition to the scaffolded MEP pathway.

In the embodiment shown in FIG. 1A and FIG. IB, a second multi-enzymatic scaffold can be co-expressed to enhance cytosolic titers of malonyl-CoA, another secondary substrate which can be used in cannabinoid biosynthesis. Such a scaffold can include an ATP citrate lyase (ACL) and acetyl-CoA carboxylase (ACC) in tandem. In some embodiments, the ACL and ACC are paired in duplicate or triplicate along the scaffold. If the ACL and ACC are paired in duplicate or triplicate, the two or three ACLs on the scaffold can be the same or different, and the two or three ACCs can be the same or different. In any of the embodiments, malonyl-CoA can be supplemented into the growth media instead of, or in addition to, being supplied by a scaffolded malonyl-CoA pathway.

In any of the embodiments in which an ACL enzyme is used, a pyruvate dehydrogenase (El) and a dihydrolipoyl transacetylase (E2) can be substituted for the ACL. For example, as shown in FIG. 4, a pyruvate dehydrogenase (El) and a dihydrolipoyl transacetylase (E2) can be substituted upstream of scaffolded mevalonate, hexanoyl-CoA, and malonyl-CoA pathways. Using both a pyruvate dehydrogenase (El) and a dihydrolipoyl transacetylase can allow acetyl-CoA to be produced using pyruvate rather than citrate as the primary substrate. In such embodiments, pyruvate also can be supplemented in the growth media. Pyruvate dehydrogenases and dihydrolipoyl transacetylases are constituents of the multi-enzyme pyruvate dehydrogenase complex

that catalyze acetyl-CoA production from pyruvate. El and E2 are found in bacteria and eukaryotes.

As shown in FIG. 1A and FIG. IB, the co-scaffolded upper cannabinoid pathway can include an olivetol synthase (OS), an olivetolic acid cyclase (OAC), and an aromatic prenyl-transferase (APT) such as a CBGA synthase (CBGAS). The upper cannabinoid pathway can begin using hexanoyl-CoA and three malonyl CoAs as the substrate for olivetol synthase-catalyzed 3,5,7-trioxododecanoyl-CoA synthesis. Olivetol synthase is classified under EC 2.3.1.206. 3,5,7-trioxododecanoyl-CoA can be used as a substrate for OAC-catalyzed olivetolic acid synthesis. OAC is classified under EC 4.4.1.26.

At the flux intersection of the converging N-terminal hexanoyl-CoA/upper cannabinoid and C-terminal mevalonate/MEP pathways (near the scaffold midpoint), an APT such as CBGAS can use olivetolic acid from the hexanoyl-CoA/upper cannabinoid pathways and geranyl pyrophosphate from the mevalonate or MEP pathway as substrates for cannabigerolate synthesis. A suitable APT is classified under EC 2.5.1.102.

In some embodiments, enzymes in the upper cannabinoid pathway can be scaffolded with a hexanoyl-CoA synthetase (HCS) to biosynthesize cannabigerolate. In some embodiments, a soluble HCS can be used with scaffolded enzymes of the upper cannabinoid pathway to biosynthesize cannabigerolate as shown in FIG. 7. Suitable enzymes for the upper cannabinoid pathway are described above.

In some embodiments, a minimal bidirectional scaffold, such as the one depicted in FIG. 8, can be used in which HCS is on the N-terminus of the scaffold, a GPPS is on the C-terminus of the scaffold, and enzymes in the upper cannabinoid pathway are scaffolded between the HCS and GPPS.

In some embodiments, such as the embodiment shown in FIG. 9, the enzymes in the upper cannabinoid pathway can be scaffolded, while the enzymes in the hexanoylC0A pathway, enzymes in the mevalonate pathway, and enzymes in the malonyl-CoA pathway can be soluble. In some embodiments, the enzymes in the upper cannabinoid pathway can be scaffolded, while the enzymes in the hexanoyl-CoA pathway, enzymes in the MEP pathway, and enzymes in the malonyl-CoA pathway can be soluble. In such embodiments, HCS can be substituted for the soluble forms of the enzymes of the

hexanoyl-CoA pathway. Suitable enzymes for each of these pathways are described above.

In some embodiments, the enzymes in the upper cannabinoid pathway can be scaffolded, while a hexanoyl-CoA synthase, enzymes in the mevalonate or MEP pathway, and enzymes in the malonyl-CoA pathway can be soluble. Suitable enzymes for each of these pathways are described above.

In some embodiments, a HCS can be scaffolded N-terminally relative to the scaffolded enzymes in the upper cannabinoid pathway, while enzymes in the mevalonate or MEP pathway, and enzymes in the malonyl-CoA pathway can be soluble. Suitable enzymes for each of these pathways are described above.

In some embodiments, the enzymes in the upper cannabinoid pathway can be scaffolded, while the enzymes in the hexanoyl-CoA pathway or a hexanoyl-CoA synthase and enzymes in the mevalonate or MEP pathways can be soluble. In some embodiments, the enzymes in the hexanoyl-CoA pathway or a hexanoyl-CoA synthase can be scaffolded N-terminal to the enzymes in the upper cannabinoid pathway, and enzymes in the mevalonate or MEP pathways can be soluble. In such embodiments, malonyl-CoA can be supplemented. Suitable enzymes for each of these pathways are described above.

In some embodiments, such as the embodiment shown in FIG. 10, a bi-directional scaffold can include enzymes of the malonyl-CoA (MCA) pathway on the N-terminus of the scaffold, enzymes of the mevalonate pathway on the C-terminus of the scaffold, and enzymes in the upper cannabinoid pathway in between. In some embodiments, a bidirectional scaffold can include enzymes of the malonyl-CoA pathway on the N-terminus of the scaffold, enzymes of the MEP pathway on the C-terminus of the scaffold, and enzymes in the upper cannabinoid pathway in between. In such embodiments, enzymes of the hexanoyl-CoA pathway can be on a separate scaffold or can be soluble. In some embodiments, HCS can be substituted for scaffolded or soluble enzymes of the hexanoylCo A pathway.

In some embodiments, each of the pathways are on separate scaffolds. For example, in one embodiment, enzymes of the upper cannabinoid pathway can be on one scaffold, enzymes of the mevalonate or MEP pathway can be localized on one scaffold,

enzymes of the hexanoyl-CoA pathway can be localized on one scaffold, and enzymes of the malonyl-CoA pathway can be localized on another scaffold.

Cannabigerolic acid biosynthesized in any of the embodiments described herein can be isolated and/or can be used as a substrate for synthesis of other secondary and tertiary cannabinoids using downstream cannabinoid synthases. In order to generate a more diverse profile of cannabinoids, the downstream cannabinoid synthases typically are not scaffolded, as scaffolding would favor production of the terminal cannabinoid. In some embodiments, however, one or more of the downstream cannabinoid synthases can be included on a scaffold described herein.

For example, one or more of cannabidiolic acid synthase (CBDAS), cannabichromenic acid synthase (CBCAS), tetrahydrocannabinolic acid synthase (THCAS), or other cannabinoid synthases can be used to produce additional cannabigerolate-derived cannabinoids. For example, a CBDAS; a CBCAS; a THCAS; a CBDAS and a CBCAS; a CBDAS and a THCAS; a CBCAS and a THCAS; or a CBDAS, CBCAS, and THCAS can be used to produce additional cannabigerolatederived cannabinoids such as one or more of cannabiodiolic acid, cannabichromenic acid, and delta-9 tetrahydrocannabinolic acid. CBDAS is classified under EC 1.21.3.8 and can catalyze the synthesis of cannabidiolic acid from cannabigerolic acid. CBCAS is classified under EC 1.3.3- and can catalyze the synthesis of cannabichromenic acid from cannabigerolic acid. THCAS is classified under EC 1.21.3.7 and can catalyze the synthesis of delta-9 tetrahydrocannabinolic acid from cannabigerolic acid.

Host cells for Producing Cannabinoids

Cannabinoids can be produced in host cells or in vitro using a multi-enzymatic scaffold as described herein. Suitable host cells include any microorganism, eukaryotic or prokaryotic, such as bacteria (e.g., Escherichia coli, Bacillus, Brevibacterium, Streptomyces, ox Pseudomonas), yeast (e.g., Pichiapastoris, Saccharomyces cerevisiae, Yarrowia lipolytica, Kluyveromyces marxianus, or Komagataellaphaffri) and other fungi (e.g., Neurospora crassa), and green algae (e.g., Dunaliella sp., Chlorella variabilis, Euglena mutabilis, or Chlamydomonas reinhardtii), as well as plant cells (e.g., tobacco,

Cannabis, or other photosynthetic plant cells) that can be maintained in culture or, in the case of plant cells such as those from tobacco or cannabis plants, can be engineered in culture and cultivated as intact transgenic plants. Such host cells or plant may or may not naturally produce cannabinoids.

A host cell can be modified to contain one or more exogenous nucleic acids that encode a scaffold as described herein and one or more exogenous nucleic acids that encode the engineered enzymes. The term “nucleic acid” as used herein encompasses both RNA and DNA, including cDNA, genomic DNA, and synthetic (e.g., chemically synthesized) DNA. The nucleic acid can be double-stranded or single-stranded. Where single-stranded, the nucleic acid can be the sense strand or the antisense strand. In addition, nucleic acid can be circular or linear.

The term “exogenous” as used herein with reference to nucleic acid and a particular host cell refers to any nucleic acid that does not originate from that particular host cell as found in nature. Thus, non-naturally-occurring nucleic acid is considered to be exogenous to a host cell once introduced into the host cell. It is important to note that non-naturally-occurring nucleic acid can contain nucleic acid sequences or fragments of nucleic acid sequences that are found in nature provided the nucleic acid as a whole does not exist in nature. For example, a nucleic acid molecule containing a genomic DNA sequence within an expression vector is non-naturally-occurring nucleic acid, and thus is exogenous to a host cell once introduced into the host cell, since that nucleic acid molecule as a whole (genomic DNA plus vector DNA) does not exist in nature. Thus, any vector, autonomously replicating plasmid, or virus (e.g., retrovirus, adenovirus, or herpes virus) that as a whole does not exist in nature is considered to be non-naturally-occurring nucleic acid. It follows that genomic DNA fragments produced by PCR or restriction endonuclease treatment as well as cDNAs are considered to be non-naturally-occurring nucleic acid since they exist as separate molecules not found in nature. It also follows that any nucleic acid containing a promoter sequence and polypeptide-encoding sequence (e.g., cDNA or genomic DNA) in an arrangement not found in nature is non-naturallyoccurring nucleic acid.

A nucleic acid that is naturally-occurring can be exogenous to a particular cell.

For example, an entire chromosome isolated from a cell of organism X is an exogenous nucleic acid with respect to a cell of organism Y once that chromosome is introduced into

Y’s cell.

It is noted that a host cell can be given an exogenous nucleic acid molecule that encodes a polypeptide having an enzymatic activity that catalyzes the production of a compound not normally produced by that host cell. Alternatively, or additionally, a host cell can be given an exogenous nucleic acid molecule that encodes a polypeptide having an enzymatic activity that catalyzes the production of a compound that is normally produced by that host cell. In this case, the recombinant host cell can produce more of the compound, or can produce the compound more efficiently, than a similar host cell not having the genetic modification.

An enzyme having a particular enzymatic activity can be a polypeptide that is either naturally-occurring or non-naturally-occurring. A naturally-occurring polypeptide is any polypeptide having an amino acid sequence as found in nature, including wild-type and polymorphic polypeptides. Such naturally-occurring polypeptides can be obtained from any species including, without limitation, animal (e.g., mammalian), plant, fungal, and bacterial species. Anon-naturally-occurring polypeptide is any polypeptide having an amino acid sequence that is not found in nature. Thus, a non-naturally-occurring polypeptide can be a mutated version of a naturally-occurring polypeptide, or an engineered polypeptide such as the engineered enzymes described herein that contain IDs. For example, a non-naturally-occurring polypeptide having geranyl pyrophosphate synthase activity can be a mutated version of a naturally-occurring polypeptide having geranyl pyrophosphate synthase activity. For example, the GPPS encoded by Erg20 may include a substitution of a tryptophan for phenylalanine at position 96 and a substitution of a tryptophan for asparagine at position 127 (referred to as Erg20ww). Erg20ww favors production of geranyl pyrophosphate over famesyl pyrophosphate. See, Jiang, etaL, Metab Eng. 2017, 41:57-66. For example, a truncated HMGR (tHMGR) such as an Nterminally truncated HMGR that includes the catalytic domain but not the transmembrane or regulatory domains of HMGR can be used. For example, the HMGR from A. thaliana

(GenBank Accession No. J04537) or a HMGR from S. cerevisiae (which contains only residues 646-1025) can be truncated to remove the transmembrane and/or regulatory domains and used in a scaffold described herein to remove a bottleneck in the mevalonate pathway. HMGR catalyzes the rate-limiting step in the mevalonate pathway (see, e.g., Song et al., 2017, Scientific reports, doi:10.1038/s41598 15005-4). For example, the nucleic acid encoding an atoB from S. cerevisiae can be modified to contain a synthetic 5’ UTR (such as the synthetic 5’ UTR sequence: 5’-cggcacccctacaaacagaaggaatataaa-3’ (SEQ ID NO:82)) and can be used in the scaffold as it alters atoB expression to facilitate flux-rebalancing in favor of production of acetoacetyl-CoA over the reverse reaction product butyryl-CoA (see Kim et al., 2018, Bioresour Technol, doi:

10.1016/j.biortech.2017.10.014). A polypeptide can be mutated by, for example, sequence additions, deletions, substitutions, or combinations thereof.

Any of the enzymes described herein that can be used to produce one or more cannabinoids can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of the corresponding wild-type enzyme. It will be appreciated that the sequence identity can be determined on the basis of the mature enzyme (e.g., with any signal sequence removed).

For example, an ACL can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a Homo sapiens ACL (see SEQ ID NO :83, FIG. 6A), or an ACL from Rattus norvegicus, Mus musculus, or Ciona intestinalis, e.g., GenBank Accession Nos. AAA74463, AAK56081, and BAB00624, respectively.

For example, an acetyl-CoA acetyltransferase (atoB) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli atoB (see SEQ ID NO:84, FIG. 6A), or an atoB from Cupriavidus necator , Clostridium acetobutylicum, or Arabidopsis thaliana , e.g., GenBank Accession Nos. CAJ92573, AAK80816, and AAM67058, respectively. In some embodiments, a malonyl-CoA acyl carrier protein transacylase from Saccharomyces cerevisiae, Homo sapiens, Serratia plymuthica, or Dickeya

paradisiaca can be substituted for atoB, e.g., GenBank Accession Nos. DAA10992,

AAH30985, AGO55277, and ACS85236, respectively.

For example, a 3-hydroxy-butyryl-CoA dehydrogenase (BHBD) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a Clostridium acetobutylicum BHBD (see SEQ ID NO:85, FIG. 6A), or a BHBD from Escherichia coli, Treponema denticola, or Arabidopsis thaliana, e.g., GenBank Accession Nos. AIZ91493, AAS11105, and AAN17431, respectively.

For example, an enoyl-CoA hydratase (ECH) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a Clostridium acetobutylicum ECH (see SEQ ID NO:86, FIG. 6A), or an ECH from Acinetobacter oleivorans, Cupriavidus necator, or Acinetobacter baumannii, e.g., GenBank Accession Nos. ADI91469, CAJ91294, and ACJ57023, respectively.

For example, a beta-ketothiolase (bktB) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a Cupriavidus necator bktB (see SEQ ID NO:87, FIG. 6A), or a bktB from Escherichia coli, Lactobacillus casei, or Clostridium acetobutylicum, e.g., GenBank Accession Nos. ALI39443, CAQ67083, and AAK80816, respectively.

For example, a trans enoyl-CoA-reductase (ECR) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a Treponema denticola ECR (see SEQ ID NO:88, FIG. 6A), or an ECR from Cupriavidus necator, Saccharomyces cerevisiae, or Klebsiella michiganensis, e.g., GenBank Accession Nos. AAP86010, DAA07148, and AIE72439, respectively.

For example, a hexanoyl-CoA synthetase (HCS), which is a type of acyl-activating enzyme (AAE), can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a C. sativa AAE1 (see SEQ ID NO:89, FIG. 6A, GenBank Accession No. AFD33345) or C. sativa AAE3 (GenBank Accession No. AFD33347). The C. sativa AAE1 and AAE3

each can use hexanoate as a substrate. See, Stout, etal., Plant J., 71(3): 353-365 (2012).

In some embodiments, the AAE encoded by CsAAEl can be used. See, GenBank

Accession No. JN717233 for the coding sequence. In some embodiments, the AAE encoded by CsAAE3 can be used. See, GenBank Accession No. JN717233 for the coding sequence. In some embodiments, both CsAAEl and CsAAE3 can be used.

For example, an HMG-C0A synthase (HMGS) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae HMGS (see SEQ ID NO :90, FIG. 6 A), or an HMGS from Arabidopsis thaliana, Lactobacillus casei, ox Homo sapiens, e.g., GenBank Accession Nos. AEE83052, CAQ67081, and AAA62411, respectively.

For example, an HMG-C0A reductase (HMGR), N-terminally truncated or canonical, can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae HMGS (see SEQ ID NO:91, FIG. 6A), or an HMGR from Arabidopsis thaliana, Lactobacillus casei, ox Homo sapiens, e.g., GenBank Accession Nos. AEE35849, CAQ67082, and AAA52679, respectively.

For example, a mevalonate kinase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae mevalonate kinase (see SEQ ID NO:92, FIG. 6A), or a mevalonate kinase from Arabidopsis thaliana, Lactobacillus casei, ox Homo sapiens, e.g., GenBank Accession Nos. AAD31719, CAQ66794, and AAF82407, respectively.

For example, a phosphomevalonate kinase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae phosphomevalonate kinase (see SEQ ID NO:93, FIG. 6A), or a mevalonate kinase from Scheffersomyces stipitis, Lactobacillus casei, or Homo sapiens, e.g., GenBank Accession Nos. EAZ63544, CAQ66339, and AAH06089, respectively.

For example, a diphosphomevalonate decarboxylase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae diphosphomevalonate decarboxylase (see SEQ

ID NO:94, FIG. 6A), or a diphosphomevalonate decarboxylase from Arabidopsis thaliana, Lactobacillus casei, or Homo sapiens, e.g., GenBank Accession Nos.

AAC67348, CAQ66795, and AAC50440, respectively.

For example, an isopentyl diphosphate isomerase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae isopentyl diphosphate isomerase (see SEQ ID NO:95, FIG. 6A), or an isopentyl diphosphate isomerase from Arabidopsis thaliana, Lactobacillus casei, or Homo sapiens, e.g., GenBank Accession Nos. AAC49920, CAQ66796, and AAP35407, respectively.

For example, a geranyl pyrophosphate synthase (GPPS) (also known as a geranyldiphospate synthase) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of the S. cerevisiae GPS or a GPPS from Acinetobacter baumannii, Lactobacillus casei, or Homo sapiens, e.g., GenBank Accession Nos. ACJ56139, CAQ66932, and AAH10004, respectively. In some embodiments, a mutant GPPS can be used. For example, the GPPS encoded by Erg20 may include a substitution of a tryptophan for phenylalanine at position 96 and a substitution of a tryptophan for asparagine at position 127 (referred to as Erg20ww) (see SEQ ID NO:96, FIG. 6A). Erg20ww favors production of geranyl pyrophosphate over farnesyl pyrophosphate. See, Jiang, et al, MetabEng. 2017 41:5766. In some cases, substituting a glutamic acid for lysine at position 179 of Erg20 (Erg20K179E) can be used to produce a GPPS that favors production of geranyl pyrophosphate. See, WO2016010827A1.

For example, a DOXP synthase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli, Clostridium acetobutylicum, Treponema denticola, or Arabidopsis thahanaDQW synthase, e.g., GenBank Accession Nos. CDH63925, AAK80036, AAS12424, and ANM65835, respectively.

For example, a DOXP reductoisomerase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli, Clostridium acetobutylicum, Treponema denticola, or

Arabidopsis thaliana DOXP reductoisomerase, e.g., GenBank Accession Nos. CDH63708, AAK79760, AAS12860, and AAM61343, respectively.

For example, a MEP cytidyl transferase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia cob, Clostridium acetobutylicum, Treponema denticola, or Arabidopsis thaliana MEP cytidyl transferase, e.g., GenBank Accession Nos. CDH66380, AAK81121, AAS12810, and BAB21592, respectively.

For example, a CDPME kinase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli, Clostridium acetobutylicum, Treponema denticola, or Arabidopsis thaliana CDPME kinase, e.g., GenBank Accession Nos. CDH64802, AAK80844, AAS11855, and AEC07908, respectively.

For example, a MECDP synthase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli, Nicotiana tabacum, Treponema denticola, or Acinetobacter baumannii MECOP synthase, e.g., GenBank Accession Nos. CDH66379, AHM22925, AAS12811, and ACJ59227, respectively.

For example, an HMBPP synthase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli, Acinetobacter baumannii, Treponema denticola, or Arabidopsis thaliana HMBPP synthase, e.g., GenBank Accession Nos. AAN81487, ACJ58210, AAS11783, and AED97354, respectively.

For example, an HMBPP reductase can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an Escherichia coli, Acinetobacter baumannii, Treponema denticola, or Arabidopsis thaliana HMBPP reductase, e.g., GenBank Accession Nos. CDH63564, ACJ57384, AAS11585, and AEE86362, respectively.

For example, an acetyl-CoA carboxylase (ACC) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a S. cerevisiae acetyl-CoA carboxylase (see SEQ ID NO:97, FIG.

6A), or an acetyl-CoA carboxylase from Homo sapiens, Treponema denticola, or

Cupriavidus necator, e.g., GenBank Accession Nos. AAP94122, AAS11086, and

CAQ67359, respectively.

For example, a pyruvate dehydrogenase (El) and dihydrolipoyl transacetylase (E2) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a Saccharomyces cerevisiae, Escherichia coli, Clostridium acetobutylicum, or Cupriavidus necator El and E2, e.g., GenBank Accession Nos. DAA07337, AMC97367, CAQ66617, and CAJ92510 for El, and DAA10474, AUG14916, CAQ66619, and CAJ92511 forE2, respectively.

For example, an olivetol synthase (OS) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an OS from C. sativa set forth in SEQ ID NO:98 (FIG. 6A) or the OS from C. sativa having GenBank Accession No. BAG14339. See, for example, Taura, et al., FEBS Letters 583 (2009) 2061-2066.

For example, an olivetolic acid cyclase (OAC) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an OAC from C. sativa set forth in SEQ ID NO :99 (FIG. 6A) or the OAC from C. sativa having GenBank Accession No. AFN42527. See, for example, Gagne, et al., Proc. Natl. Acad. Sci. USA, 2012 109 (31) 12811-12816.

For example, a CBGAS can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of an aromatic prenyl-transferase (APT) from Cannabis sativa such as the CBGAS set forth in SEQ ID NO: 100 (FIG. 6A). See, for example, U.S. Patent Publication No. 20120144523A1 and U.S. Patent No. 8,884,100B2. In some embodiments, a soluble APT from Streptomyces (e.g., NphB) can be used. See, for example, Carvalho et al., FEMS Yeast Research, 17, 2017, fox037.

For example, a cannabidiolic acid synthase (CBDAS) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a CBDAS from C. sativa set forth in SEQ ID NO: 101 (FIG.

6A) or the amino acid sequence of a CBDAS from C. sativa having GenBank Accession

No. BAF65033. See, for example, Taura, et al.,FEBS Lett. 581 (16), 2929-2934 (2007).

For example, a cannabichromenic acid synthase (CBCAS) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a CBCAS from C. sativa set forth in SEQ ID NO: 102 (FIG. 6A) or the amino acid sequence of a CBCAS from C. sativa as set forth in SEQ ID NO:2 of WO 2015/196275 Al. SEQ ID NO:2 of WO 2015/196275 Al includes anN-terminal 28 amino acid signal peptide. All or a portion of the signal peptide can be removed from the sequence. The CBDAS from C. indica or C. ruderalis also can be used. In some embodiments, an Escherichia coli or yeast optimized nucleic acid sequence encoding a C. sativa CBCAS as set forth in SEQ ID NOs: 8 and 9, respectively, of WO 2015/196275 Al can be used.

For example, a tetrahydrocannabinolic acid synthase (THCAS) can have at least 70% sequence identity (e.g., at least 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, or 100%) to the amino acid sequence of a THCAS from C. sativa having GenBank Accession No. BAC41356. See, for example, Sirikantaramas, etal., J. Biol. Chem. 279 (38), 39767-39774 (2004).

The percent identity (homology) between two amino acid sequences can be determined as follows. First, the amino acid sequences are aligned using the BLAST 2 Sequences (B12seq) program from the stand-alone version of BLASTZ containing BLASTP version 2.0.14. This stand-alone version of BLASTZ can be obtained from Fish &amp; Richardson’s web site (e.g., www.fr.com/blast/) or the U.S. government’s National Center for Biotechnology Information web site (www.ncbi.nlm.nih.gov). Instructions explaining how to use the B12seq program can be found in the readme file accompanying BLASTZ. B12seq performs a comparison between two amino acid sequences using the BLASTP algorithm. To compare two amino acid sequences, the options of B12seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C:\seql.txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -0 is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default setting. For example, the following command

can be used to generate an output file containing a comparison between two amino acid sequences: C:\B12seq-i c:\seql.txt-j c:\seq2.txt-p blastp-0 c:\output.txt. If the two compared sequences share homology (identity), then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology (identity), then the designated output file will not present aligned sequences. Similar procedures can be following for nucleic acid sequences except that blastn is used.

Once aligned, the number of matches is determined by counting the number of positions where an identical amino acid residue is presented in both sequences. The percent identity (homology) is determined by dividing the number of matches by the length of the full-length polypeptide amino acid sequence followed by multiplying the resulting value by 100. It is noted that the percent identity (homology) value is rounded to the nearest tenth. For example, 78.11, 78.12, 78.13, and 78.14 is rounded down to 78.1, while 78.15, 78.16, 78.17, 78.18, and 78.19 is rounded up to 78.2. It also is noted that the length value will always be an integer.

It will be appreciated that a number of nucleic acids can encode a polypeptide having a particular amino acid sequence. The degeneracy of the genetic code is well known to the art; i.e., for many amino acids, there is more than one nucleotide triplet that serves as the codon for the amino acid. For example, codons in the coding sequence for a given enzyme can be modified such that optimal expression in a particular species (e.g., bacteria or fungus) can be attained, using appropriate codon bias tables for that species. For example, the nucleotide sequences set forth in FIG. 12A are the nucleic acid sequences encoding an ATP citrate lyase, an atoB, a 3-hydroxbutyryl-CoA dehydrogenase, an enoyl-CoA hydratase, a beto-ketothiolase (bktB), a trans-enoyl-CoA reductase, an HMG-C0A synthase, an HMG-C0A reductase, a mevalonate kinase, a phosphomevalonate kinase, a diphosphomevalonate decarboxylase, an isopentenyldiphosphate delta isomerase, a geranyl-diphosphate synthase (ERG20ww), an olivetol synthase, an olivetolic acid cyclase, a CBGA synthase, a CBDA synthase, a CBCA synthase, an acetyl-CoA carboxylase, and a hexanoyl-CoA synthetase. The nucleic acid sequences for the ATP citrate lyase, atoB, 3-hydroxybutyryl-CoA dehydrogenase, enoyl48

C0A hydratase, trans-enoyl-CoA reductase, bktB, olivetol synthase, olivetolic acid cyclase, CBGA synthase, CBDA synthase, and CBCA synthase have been codon optimized for expression in yeast. FIGs. 14A-14C contain codon optimized (for expression in yeast) nucleic acid sequences encoding the engineered enzymes of FIGs.

13A-13C.

In addition to sequence similarity, it will be appreciated that enzymes and scaffolds with structural and/or functional similarity to the enzymes and scaffolds described herein are also encompassed within the scope of the document.

This document provides recombinant host cells that can be used to produce one or more cannabinoids as described herein. For example, an individual host cell can contain exogenous nucleic acid such that the scaffold polypeptide and each of the enzymes to be immobilized on the scaffold are expressed. It is important to note that such host cells can contain any number and/or combination of exogenous nucleic acid molecules. For example, a particular host cell can contain an exogenous nucleic acid encoding the scaffold, and additional exogenous nucleic acids encoding the enzymes of the malonylC0A pathway, enzymes of the hexanoyl-CoA pathway or encoding a HCS, and enzymes of the mevalonate or MEP pathway. A single exogenous nucleic acid can encode one enzyme or more than one enzyme (e.g., one or more copies of from one to ten (or more) enzymes, from one to eight, from one to seven, from one to six, from one to five, from one to four, or from two to three enzymes). Thus, the number of different exogenous nucleic acids needed to produce the engineered enzymes to be localized on the scaffold will depend on the design of the scaffold and/or the particular embodiment. FIG. 2A and FIG. 2B each provide a non-limiting schematic of suitable gene cassettes for expressing the scaffolds and enzymes. FIG. 12C provides the nucleic acid sequence encoding a scaffold polypeptide containing the peptide ligands corresponding to IDs 1-16 as shown in Table 2 and a triplicate MYC tag. See also FIG. 14D for the codon-optimized nucleic acid sequence encoding the scaffold polypeptide of FIG. 13D . FIG. 12D provides the nucleic acid sequence encoding a scaffold polypeptide that contains the peptide ligands corresponding to IDs 1 and 17, and a triplicate FLAG tag. See also FIG. 14D.

In some embodiments, multiple nucleic acids encoding polypeptides (e.g., the nucleic acids of a gene cassette such as in FIG. 2A or FIG. 2B) can be linked together using a nucleic acid sequence encoding a self-cleaving peptide. During translation of the transcripts, the growing polypeptide can be cleaved at the 2A peptide with translation continuing through to the next polypeptide. When designing a vector to express the polypeptides as a polycistronic unit, the nucleic acid encoding the polypeptides and the self-cleaving peptide (e.g., a 2A peptide) can be designed such that they are in translational frame with each other. Examples of 2A peptides that can be used as described herein include, without limitation, a 2A peptide of foot-and-mouth disease virus (FMDV), a 2A peptide of equine rhinitis A virus (ERAVO), a 2A peptide of Thosea asigna virus (TaV), or a 2A peptide of porcine teschovirus-1 (PTV-1) or porcine teschovirus-2 (PTV-2). The 2A peptides from PTV-1 and PTV-2 are referred to as P2A peptides. See, e.g., SEQ ID NO:212 for a codon-optimized nucleotide sequence (for S. cerevisiae) encoding a P2A peptide.

Further, the cells described herein can contain a single copy or multiple copies (e.g., about 5, 10, 20, 35, 50, 75, 100 or 150 copies), of a particular exogenous nucleic acid molecule. Again, the cells described herein can contain more than one particular exogenous nucleic acid molecule and/or copies thereof. For example, a particular cell can contain about 50 copies of exogenous nucleic acid molecule X as well as about 75 copies of exogenous nucleic acid molecule Y.

Any method can be used to introduce an exogenous nucleic acid molecule into a host cell. In fact, many methods for introducing nucleic acid into host cells such as bacteria and yeast are well known to those skilled in the art. For example, heat shock, lipofection, electroporation, nucleofection, conjugation, fusion of protoplasts, and biolistic delivery are common methods for introducing nucleic acid into bacteria and yeast cells. See, e.g., Ito etaL, J. BacteroL 153:163-168 (1983); Durrens et aL, Curr Genet. 18:7-12 (1990); and Becker and Guarente, Methods in Enzymology 194:182-187 (1991).

An exogenous nucleic acid molecule contained within a particular host cell can be maintained within that host cell in any form. For example, exogenous nucleic acid

molecules can be integrated into the genome of the microorganism or maintained in an episomal state. In other words, a microorganism can be a stable or transient transformant.

Again, a microorganism described herein can contain a single copy, or multiple copies (e.g., about 5, 10, 20, 35, 50, 75, 100 or 150 copies), of a particular exogenous nucleic acid molecule as described herein.

Suitable nucleic acid constructs for expressing the engineered enzymes and scaffolds include, for example, CRISPR plasmids, baculovirus vectors, bacteriophage vectors, plasmids, phagemids, cosmids, fosmids, bacterial artificial chromosomes, viral vectors (for example, viral vectors based on vaccinia virus, poliovirus, adenovirus, adeno-associated virus, SV40, herpes simplex virus, and the like), Pl-based artificial chromosomes, yeast plasmids, yeast artificial chromosomes, and other vectors. Typically such constructs include a regulatory element that promotes the expression of a nucleic acid sequence that encodes a polypeptide. Typically, regulatory elements are DNA sequences that regulate the expression of other DNA sequences at the level of transcription. Thus, regulatory elements include, without limitation, promoters, enhancers, and the like. Any type of promoter can be used to express an amino acid sequence from an exogenous nucleic acid molecule. Examples of promoters include, without limitation, constitutive promoters, tissue-specific promoters, and inducible or repressible promoters that are responsive or unresponsive to a particular stimulus (e.g., light, oxygen, chemical concentration, sound, and the like).

In some embodiments, endogenous yeast promoters with varying constitutive activity levels can be used to express the engineered enzymes and/or scaffolds. To maintain an excess of enzymes relative to scaffold molecules, the scaffolds can be expressed under control of the weakest promoter. For example, one or more of the following yeast promoters can be used: the promoter from the gene encoding transcriptional elongation factor EF-1 a (pTEFl), the promoter from the gene encoding phosphoglycerate kinase (PGK1), the promoter from the gene encoding triose phosphate isomerase (pTPIl), the promoter from the gene encoding a hexose transporter (pHXT7), HXT7, the promoter from the gene encoding pyruvate kinase 1 (pPYKl), the promoter from the gene encoding alcohol dehydrogenase 1 (pADHl), or the promoter from the

gene encoding triphosphate dehydrogenase (pTDH3). For example, in the embodiment shown in FIG. 2 A, the pTPIl promoter can be used to express enzymes of the upper hexanoyl-CoA (HCA), enzymes of the lower HCA pathway, enzymes of the upper mevalonate (MVA) pathway, enzymes of the lower MVA pathway, and enzymes of the lower cannabinoid (CB) pathway, while the pTEFl promoter can be used to express enzymes of the upper CB pathway, the atoB enzyme, and the enzymes of the malonylC0A pathway, and the pADHl promoter can be used to express the scaffold. Of these promoters, the pADHl promoter has the weakest activity (+ in FIG. 2A), the pTEFl promoter has the strongest activity (+++ in FIG. 2A), and the activity of the pTPIl promoter is between the other two (++ in FIG. 2A). In some embodiments, the Gal 1-10 promoter (e.g., from S. cerevisiae) can be used. See, e.g., FIG. 17.

A nucleic acid construct also can include a selectable marker, e.g., for an antibiotic such as neomycin resistance, ampicillin resistance, tetracycline resistance, chloramphenicol resistance, or kanamycin resistance). In some embodiments, a nutritional marker gene that confers prototrophy for an essential nutrient such as tryptophan (TRP1), uracil (URA3), histidine (HIS3), leucine (LEU2), lysine (LYS2), or methionine can be included on a nucleic acid construct. See, e.g., FIG. 17. As shown in Example 3, four different auxotrophic markers were used to sequentially select for transformed cells containing the desired combinations of nucleic acids encoding the enzymes and scaffold. For example, yeast cells transformed with a vector containing a TRP gene and the nucleic acids encoding enzymes of the hexanoyl-CoA pathway were grown in tryptophan deficient media. The transformed cells that grew in the tryptophan deficient media were selected and further transformed with a vector containing a LEU gene and nucleic acid encoding enzymes of the mevalonate pathway. The resulting transformed cells were grown on media lacking tryptophan and leucine, and the cells that grew in the media lacking tryptophan and leucine were transformed with a vector containing a HIS gene and nucleic acids encoding enzymes of the upper cannabinoid pathway. The resulting transformed cells were grown on media lacking tryptophan, leucine, and histidine, and the cells that grew in the media lacking tryptophan, leucine, and histidine were transformed with a vector containing a URA3 gene and a nucleic acid

encoding a scaffold. The resulting transformed cells were grown on media lacking tryptophan, leucine, histidine, and uracil. Cells that grew in media lacking tryptophan, leucine, histidine, and uracil contained the desired combination of enzymes and scaffold as shown in FIG. IB.

In some embodiments, the encoded enzymes (e.g., one or more enzymes from the cannabinoid biosynthesis pathway, mevalonate pathway, MEP pathway, hexanoyl-CoA pathway, or a hexanoyl-CoA synthetase) and/or the scaffold can include a targeting sequence that can be used to direct the enzymes or scaffold to one of several different intracellular compartments, including, for example, the endoplasmic reticulum (ER), mitochondria, plastids (such as chloroplasts), the vacuole, the Golgi apparatus, or protein storage vesicles (PSV). For example, a mitochondrial or plastidial targeting sequence can be used to facilitate mitochondrial or plastidial compartmentalization of cannabinoid/cannabinoid precursor biosynthesis such that the encoded enzymes and scaffold are expressed in the mitochondria or plastids of the host cell.

In some embodiments, cannabinoid/cannabinoid precursor biosynthesis can be performed in two compartments by co-expressing one or more engineered enzymes and a scaffold in both the cytosolic compartment and either the plastids or mitochondria of the host cell. See, for example, FIG. 11. It will be appreciated that while FIG. 11 depicts a scaffold containing enzymes of the hexanoyl-CoA pathway, enzymes of the upper cannabinoid pathway, and enzymes of the mevalonate pathway, dual-compartment engineering can be performed with any of the scaffolds and enzymes described herein. For example, dual-compartment engineering can be performed in two compartments by co-expressing a scaffold and enzymes of the hexanoyl-CoA pathway, enzymes of the upper cannabinoid pathway, and enzymes of the MEP pathway in both the cytosolic compartment and either the plastids of mitochondria of the host cell. Dual-compartment engineering also can be achieved by engineering separate haploid yeast strains for cytosolic and mitochondrial/plastidial cannabinoid biosynthesis, and then mating these two haploid strains to produce a diploid lineage that is heterozygous for cytosolic and mitochondrial/plastidial cannabinoid biosynthesis.

In some embodiments, the engineered enzymes and/or scaffolds also contain a tag that can be used for purification of the recombinant protein (e.g., c-myc, FLAG; polyhistidine (e.g., hexahistidine), hemagglutinin (HA), glutathione-S-transferase (GST), or maltose binding protein (MBP)) or as a detectable marker (e.g., luciferase, green fluorescent protein (GFP), or chloramphenicol acetyl transferase (CAT)). For example, in the embodiment shown in FIG. 6C and FIG. 6D, a scaffold can include a myc tag (e.g., (Myc)3 tag) or a FLAG tag (FLAG)3 tag at the C-terminus.

In some embodiments, a host cell can be engineered to increase acetyl-CoA availability for cannabinoid and cannabinoid precursor biosynthesis. For example, the mitochondrial enzyme isocitrate dehydrogenase-1 (IDH1) can be placed under transient micro-RNA-mediated inducible repression. Since mitochondrial IDH1 is primarily responsible for depletion of the cellular citrate pool, micro-RNA-mediated repression of IDH1 can increase the availability and cytosolic shuttling of citrate for production of acetyl-CoAby ATP citrate lyase. The resulting increase in acetyl-CoA bioavailability can further enhance downstream hexanoyl-CoA and geranyl pyrophosphate titers by improving initial substrate availability for the hexanoyl-CoA and mevalonate pathways. The combinatorial metabolic engineering of acetyl-CoA can mitigate issues related to the siphoning of acetyl-CoA away from the endogenous metabolism of the host cells.

In some embodiments, one or more conventional and/or contemporary gene editing techniques can be used to produce recombinant hosts. For example, clustered, regularly interspaced, short palindromic repeat (CRISPR) technology can be used to modify expression of an endogenous nucleic acid. The CRISPR/Cas system includes components of a prokaryotic adaptive immune system that is functionally analogous to eukaryotic RNA interference, using RNA base pairing to direct DNA or RNA cleavage. The Cas9 protein functions as an endonuclease, and CRISPR RNA (crRNA) and transactivating RNA (tracrRNA) sequences complex with the Cas9 enzyme and direct it to a target DNA sequence (Makarova et al., Nat Rev Microbiol 9(6):467-477, 2011). The modification of a single targeting RNA can be sufficient to alter the nucleotide target of a Cas protein. In some cases, crRNA and tracrRNA can be engineered as a single cr/tracrRNA hybrid (also referred to as a “guide RNA” or “gRNA”) to direct Cas9

cleavage activity (Jinek et al., Science, 337(6096):816-821, 2012). The CRISPR/Cas system can be used in a variety of prokaryotic and eukaryotic organisms (see, e.g., Jiang et al., Nat Biotechnol, 31(3):233-239, 2013; Dicarlo et al., Nucleic Acids Res, doi:10.1093/nar/gktl35, 2013; Cong et al., Science, 339(6121):819-823, 2013; Mali et al., Science, 339(6121):823-826, 2013; Cho et al., Nat Biotechnol, 31(3):230-232, 2013; and Hwang et al., Nat Biotechnol, 31(3):227-229, 2013).

Another gene-editing technique can include a sequence-specific nuclease created by fusing transcription activator-like effectors (TALEs) to, for example, the catalytic domain of the Fold endonuclease. Both native and custom TALE-nuclease (“TALEN”) fusions direct DNA double-strand breaks to specific, targeted sites. See, for example, Christian, etaL, Genetics 186: 757-761 (2010) and U.S. Patent Publication No. 20110145940.

Other suitable gene insertion techniques include the use of retroviral vectors and biolistic particle gene delivery systems (colloquially known as “gene guns”).

Methods of identifying and/or selecting host cells that contain exogenous nucleic acid or a modified endogenous nucleic acid are well known to those skilled in the art. Such methods include, without limitation, the introduction and expression of a negative selection marker such as an antibiotic resistance gene, PCR, and nucleic acid hybridization techniques such as Northern and Southern analyses. In some cases, immunohistochemistry and biochemical techniques can be used to determine if a microorganism contains a particular nucleic acid by detecting the expression of the encoded enzymatic polypeptide encoded by that particular nucleic acid molecule. For example, an antibody having specificity for an encoded enzyme can be used to determine whether or not a particular cell contains that encoded enzyme. Further, biochemical techniques can be used to determine if a cell contains a particular nucleic acid molecule encoding an enzymatic polypeptide by detecting an organic product produced as a result of the expression of the enzymatic polypeptide.

This document also provides isolated nucleic acids molecules. The term “isolated” as used herein with reference to nucleic acid refers to a naturally-occurring nucleic acid that is not immediately contiguous with both of the sequences with which it is

immediately contiguous (one on the 5' end and one on the 3' end) in the naturallyoccurring genome of the organism from which it is derived. For example, an isolated nucleic acid can be, without limitation, a recombinant DNA molecule of any length, provided one of the nucleic acid sequences normally found immediately flanking that recombinant DNA molecule in a naturally-occurring genome is removed or absent. Thus, an isolated nucleic acid includes, without limitation, a recombinant DNA that exists as a separate molecule (e.g., a cDNA or a genomic DNA fragment produced by PCR or restriction endonuclease treatment) independent of other sequences as well as recombinant DNA that is incorporated into a vector, an autonomously replicating plasmid, a virus (e.g., a retrovirus, adenovirus, or herpes virus), or into the genomic DNA of a prokaryote or eukaryote. In addition, an isolated nucleic acid can include a recombinant DNA molecule that is part of a hybrid or fusion nucleic acid sequence.

The term “isolated” as used herein with reference to nucleic acid also includes any non-naturally-occurring nucleic acid since non-naturally-occurring nucleic acid sequences are not found in nature and do not have immediately contiguous sequences in a naturally-occurring genome. For example, non-naturally-occurring nucleic acid such as an engineered nucleic acid is considered to be isolated nucleic acid. Engineered nucleic acid can be made using common molecular cloning or chemical nucleic acid synthesis techniques. Isolated non-naturally-occurring nucleic acid can be independent of other sequences, or incorporated into a vector, an autonomously replicating plasmid, a virus (e.g., a retrovirus, adenovirus, or herpes virus), or the genomic DNA of a prokaryote or eukaryote. In addition, a non-naturally-occurring nucleic acid can include a nucleic acid molecule that is part of a hybrid or fusion nucleic acid sequence.

It will be apparent to those of skill in the art that a nucleic acid existing among hundreds to millions of other nucleic acid molecules within, for example, cDNA or genomic libraries, or gel slices containing a genomic DNA restriction digest is not to be considered an isolated nucleic acid.

In some embodiments, the production of one or more cannabinoids can be performed in vitro using the scaffold and immobilized enzymes described herein, using a lysate (e.g., a buffered cell lysate) from a recombinant host cell as a source of the scaffold

and enzymes, using a plurality of lysates from different host cells as the source of the scaffold and enzymes, or using an acellular reaction buffer such as a synthetic reaction buffer. For example, following co-immunoprecipitation of C-terminal Myc/Flag-tagged enzyme-bound scaffolds, scaffold-enzyme complexes can be maintained in a citratesupplemented and/or glucose-supplemented (or other carbon source-supplemented) reaction buffer which allows in-vitro scaffolded cannabinoid biosynthesis.

Producing Cannabinoids Using a Recombinant Host

Typically, one or more cannabinoids can be produced by providing a recombinant host such as a recombinant microorganism and culturing the microorganism with a culture medium. In general, the culture media and/or culture conditions can be such that the microorganisms grow to an adequate density and produce cannabinoids efficiently. For example, the microorganisms can be subjected to aerobic batch fermentation. In some embodiments, one or more precursors (e.g., citrate, glucose, hexanoic acid, and/or other carbon source and/or malonyl-CoA) are supplemented in the culture medium. In some embodiments, about 30 mg/L to about 10,000 mg/L (e.g., about 100 mg/L to about 5,000 mg/L, about 200 mg/L to about 4,000 mg/L, about 300 mg/L to about 3,000 mg/L, or about 350 mg/L to about 1,000 mg/L) of buffered citrate, pH 6.0 can be added to the culture medium.

For large-scale production processes, any method can be used such as those described elsewhere (Manual of Industrial Microbiology and Biotechnology, 2nd Edition, Editors: A. L. Demain and J. E. Davies, ASM Press; and Principles of Fermentation Technology, P. F. Stanbury and A. Whitaker, Pergamon). Briefly, a large vessel (e.g., a 100 gallon, 200 gallon, 500 gallon, or higher volume vessel) containing an appropriate culture medium is inoculated with a particular microorganism. After inoculation, the microorganism is incubated to allow biomass to be produced. Once a desired biomass or cellular confluency is attained, a portion or all of the broth containing the microorganisms can be transferred to a second vessel. This second vessel can be any size. For example, the second vessel can be larger, smaller, or the same size as the first vessel. Typically, the second vessel is larger than the first such that additional culture medium can be added to

the broth from the first vessel. In addition, the culture medium within this second vessel can be the same as, or different from, that used in the first vessel. This system can expand to include an array consisting of any number of individual vessels.

Once transferred, the microorganisms can be incubated to allow for the production of one or more cannabinoids. Once produced, any method can be used to isolate cannabinoids. For example, common separation techniques can be used to remove the biomass from the broth, and common isolation procedures (e.g., extraction such as non-polar extraction with hexane followed by ethyl-acetate), high-performance liquid chromatography (e.g., HPLC with a diode array detector (HPLC-DAD)), gas chromatography-flame ionization detection (GC-FID), or ion-exchange procedures) can be used to obtain the cannabinoids from the biomass.

A host cell described herein can produce one or more cannabinoids at a concentration of at least about 10 mg per L (e.g., at least about 15 mg/L 25 mg/L, 50 mg/L, 75 mg/L, 100 mg/L, 150 mg/L, 200 mg/L, 250 mg/L or more). For example, in some embodiments, total cannabinoids (total of CBG; CBGA, CBD, CBDA, CBC, and CBCA) can be produced at a concentration of at least about 10 mg/L, 15 mg/L, 20 mg/L, 40 mg/L, 60 mg/L, 80 mg/L, or 100 mg/L or more. For example, in some embodiments, total cannabinoids (total of CBG; CBGA, CBD, CBDA, CBC, and CBCA) can be produced at a concentration from about 10 mg/L to about 500 mg/L (e.g., 20 mg/L to 450 mg/L, 40 mg/L to 380 mg/L, 60 mg/L to 280 mg/L, 60 mg/L to 250 mg/L, 60 mg/L to 150 mg/L, 80 mg/L to 400 mg/L, 80 mg/L to 300 mg/L, 80 mg/L to 250 mg/L, 80 mg/L to 200 mg/L, 80 mg/L to 175 mg/L, 90 mg/L to 400 mg/L, 90 mg/L to 300 mg/L, 90 mg/L to 250 mg/L, or 90 mg/L to 150 mg/L). In some embodiments, one or more individual cannabinoids (e.g., one or more of CBG; CBGA, CBD, CBDA, CBC, and CBCA) can be produced at concentrations of at least about 1 mg/L, 2 mg/L, 5 mg/L, 10 mg/L, 15 mg/L, 20 mg/L, 25 mg/L, 30 mg/L, 35 mg/L, 40 mg/L, 45 mg/L, 50 mg/L, 55 mg/L, 60 mg/L, 65 mg/L, 70 mg/L, 75 mg/L, 80 mg/L, 85 mg/L, 90 mg/L, 95 mg/L, 100 mg/L or more. For example, in some embodiments, one or more individual cannabinoids can be produced at a concentration from about 1 mg/L to about 100 mg/L (e.g., 2 to 90 mg/L, 2 to 80 mg/L, 2 to 70 mg/L, 2 to 60 mg/L, 2 to 50 mg/L, 2 to 40 mg/L, 2 to 30

mg/L, 2 to 20 mg/L, 2 to 15 mg/L, 3 to 90 mg/L, 3 to 80 mg/L, 3 to 70 mg/L, 3 to 60 mg/L, 3 to 50 mg/L, 3 to 40 mg/L, 3 to 30 mg/L, 3 to 20 mg/L, 3 to 15 mg/L, 4 to 90 mg/L, 4 to 80 mg/L, 4 to 70 mg/L, 4 to 60 mg/L, 4 to 50 mg/L, 4 to 40 mg/L, 4 to 30 mg/L, 4 to 20 mg/L, or 4 to 15 mg/L).

The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

EXAMPLES

Example 1 - General Methods

Enzymatic Constructs

Each enzyme construct is designed to include an interaction domain (ID) which is comprised of two tandem N-terminal or C-terminal ligand-binding motifs which are separated from the given enzyme and from one another by an amino acid sequence containing flexible GS-rich linkers flanking a rigid a-helical spacer sequence. The motifs comprising the ID of each enzyme specifically bind tandem peptide ligands which form ID-binding sites at discrete locations along a synthetic intracellular polypeptide scaffold. Expression of each enzyme is controlled by a constitutive or inducible promoter. The nucleic acid encoding the enzyme can be codon optimized, e.g., for expression in yeast.

Scaffolding Constructs

ID-binding sites containing tandem peptide ligands that are specific for the tandem scaffold-binding motifs, which comprise the ID of each enzyme, are inserted at discrete positions along an intracellular polypeptide scaffold.

The tandem ligands which comprise each scaffolded ID-binding site are separated from one another by a 36 amino acid residue sequence containing flexible GS-rich linkers flanking a rigid a-helical spacer sequence, while the scaffolded ID-binding sites themselves are separated from one another by a 50 amino acid residue sequence (or any other number of amino acid residues) containing flexible GS-rich linkers flanking a rigid a-helical spacer sequence. Specifically, the scaffold binding sites for each enzyme in the hexanoyl-CoA pathway are positioned (in order of catalysis) proximally to ATP citrate

lyase and acetyl-CoA acetyltransferase at the N-terminus of the primary scaffold. Scaffold binding sites for each enzyme in the upper cannabinoid pathway are positioned proximally to (immediately downstream of) the binding sites for the hexanoyl-CoA pathway enzymes. The scaffold binding sites for each enzyme in the mevalonate (or MEP) pathway are positioned (in order of catalysis) proximally to ATP citrate lyase and acetyl-CoA acetyltransferase at the C-terminus of the primary scaffold. The enzyme catalyzing the rate-limiting/committed step in cannabinoid biosynthesis (CBGA synthase, the final enzymatic step in the upper cannabinoid pathway) is located at the intersection of the converging cannabinoid precursor pathways near the scaffold midpoint.

Assessment of Cannabinoidergic Potential by Transient Transfection

Competent yeast and/or green algae cells are transiently transfected with plasmids encoding various permutations of the scaffold and enzymes. To establish baseline cannabinoidergic capacity, cells first undergo transient transfection with the enzymes required for cannabinoid biosynthesis (but not the scaffolds), and biosynthesized cannabinoids are extracted, isolated, and quantified as described below (see “Cannabinoid Extraction, Isolation, and Analytical Characterization”). To measure the improvement in cannabinoidergic capacity conferred by multi-enzymatic scaffolding, a subset of the aforementioned cells is co-transfected with plasmids encoding one or more of the multi-enzymatic scaffolds described herein, and biosynthesized cannabinoids are extracted, isolated, and quantified. The presence of the plasmid DNA is confirmed by PCR, functional gene expression is confirmed by qRT-PCR, protein/polypeptide production is confirmed by Western blotting, and scaffolding of each enzyme is confirmed by co-immunoprecipitation of C-terminal myc/flag-tagged scaffolds followed by Western blot analysis of each co-immunoprecipitated enzyme.

Engineering of Stable Cannabinoidergic Cell Lines

The constructs can be integrated into the genome of host cells such yeast, green algae, or other suitable hosts via stable transfection. Gene integration is confirmed by PCR, functional gene expression is confirmed by qRT-PCR, and protein/polypeptide

production is confirmed by Western blotting. Gene expression/protein synthesis is confirmed by comparing both qRT-PCR and Western blot results among samples with and without genetic engineering. To assess the improvement in cannabinoidergic capacity conferred by multi-enzymatic scaffolding for stably engineered cannabinoidergic cell lines, cannabinoid biosynthesis will be compared among cells that are stimulated for enzyme but not scaffold expression and cells that are stimulated for enzyme and scaffold expression.

Validation of Multi-enzymatic Scaffolding

To verify successful multi-enzymatic scaffolding in both transiently transfected and stably engineered cells, a myc-tag (or other immunoprecipitable tag) is inserted at the N-terminal or C-terminal of the polypeptide scaffold(s). Scaffolded enzymes are selectively co-immunoprecipitated by affinity chromatography using anti-myc affinity beads. Western blots are performed to detect and quantify each co-immunoprecipitated enzyme.

Aerobic Fed-batch Fermentation

Stably engineered cannabinoidergic yeast, green algae, or other host cells are grown in bioreactors (or any other vessel) via aerobic batch fermentation (or any other culture technique).

Cannabinoid Extraction, Isolation, and Analytical Characterization

Following sufficient elicitation of cannabinoid biosynthesis, engineered yeast/green algae cells are pelleted by centrifugation and washed with TBS. The supernatant (liquid culture media) is decanted and collected. Following washing with TBS, pelleted cells are resuspended in NaOH adjusted ethanol and lysed by iterative freeze-thawing and ultrasonication. Biosynthesized cannabinoid fermentates are then harvested from both lysates and supernatants via triplicate nonpolar extractions using hexane followed by ethyl-acetate. The resulting organic fractions are pooled and rotoevaporated. High-performance liquid chromatography with a diode array detector

(HPLC-DAD) or gas chromatography-flame ionization detection (GC-FID) is then applied for quantitative and qualitative measurement of biosynthesized cannabinoids.

In the following examples, each 48-hour culture was lysed/homogenized by ultrasonication. Ultrasonicated samples were then subjected to triplicate liquid-liquid extractions with ethyl acetate (one volumetric equivalent of ethyl acetate per extraction). Following separation, the ethyl acetate fractions collected from each sample were pooled, and the pooled samples were centrifugally filtered. Ethyl acetate was then removed from each sample in a vacuum oven, and the residual samples were resuspended in 10mL methanol for analytical characterization. Analytical characterization of all samples was conducted by a licensed, independent, third-party analytical testing facility (Precision Plant Molecules, Denver, CO). HPLC-DAD was utilized for quantitative and qualitative measurement of each parent and derivative cannabinoid as well as the cannabinoid precursor OVA.

Example 2 - Synthetic Gene Cassette Assembly/Synthesis, Plasmid Preparation, and Polycistronic Vector Construction

Five synthetic gene cassettes (entitled HCA, GPP, CAN, SCF, and SOL) were constructed for biosynthesizing cannabinoids in heterologous cells or acellular reaction buffers. See, Figure 2B. The cassettes collectively encode all scaffold-binding engineered enzymes and the polypeptide scaffolds to which the engineered enzymes can bind.

The HCA gene cassette encoded scaffold-binding engineered enzymes for scaffolded hexanoyl-CoA biosynthesis, namely ACL, atoB, BHBD, ECH, ECR, and bktB, and encoded a soluble HCS for additional hexanoyl-CoA production from hexanoate-supplemented culture media or acellular reaction buffer. See, FIG. 13 A. The GPP gene cassette encoded scaffold-binding engineered enzymes for scaffolded geranyl pyrophosphate (GPP) biosynthesis, namely HMGS, tHMGR, ERG12, ERG8, MVD1, IDI1, and ERG20ww. See, FIG. 13B. The CAN gene cassette encoded scaffold-binding engineered enzymes for scaffolded OAC, malonyl-CoA, and CBGA biosynthesis, namely OS and OAC, ACC, and CBGAS, respectively, as well all enzymes for soluble (non62

scaffolded) CBDA and CBCA biosynthesis, namely CBDAS and CBCAS, respectively. See, FIG. 13C. The SCF gene cassette encoded the polypeptide scaffolds for bidirectional scaffolded cannabinoid biosynthesis and scaffolded malonyl-CoA biosynthesis, namely the cannabinoidergic metabolon scaffold (CBSCF) and the malonyl-CoA metabolon scaffold (MCASCF), respectively, as well as additional copies of both ACL and atoB to enhance acetyl-CoA biosynthesis from supplemental and/or endogenous citrate and acetoacetyl-CoA biosynthesis from acetyl-CoA, respectively. See, FIG. 13D. The SOL gene cassette lacked the polypeptide scaffolds for bidirectional scaffolded cannabinoid biosynthesis and scaffolded malonyl-CoA biosynthesis (i.e., it was used for soluble cannabinoid biosynthesis) but, analogous to the SCF gene cassette, encoded additional copies of ACL and atoB to enhance acetyl-CoA biosynthesis from supplemental and/or endogenous citrate and acetoacetyl-CoA biosynthesis from acetyl-CoA. See FIG. 13 A for the amino acids sequences of the ACL and atoB engineered enzymes.

Gene cassettes were assembled/ synthesized using self-cleaving 2A peptides (P2As) to link multiple codon-optimized (for S. cerevisiae) gene sequences assigned to each cassette. To improve P2A cleavage, a GSG linker (comprised of a single serine residue flanked by single glycine residues) was inserted at the interface between each constituent gene sequence and the P2A linker sequence to which it was fused (of the format: gene cassette sequence 1 - SG - P2A linker - gene cassette sequence 2 - GSG P2A linker - gene cassette sequence 3 - GSG - P2A linker -) and so forth. See, FIGs. 14A-14D for codon-optimized nucleic acid sequences encoding the engineered enzymes and scaffolds. Following assembly, each synthetic gene cassette was inserted into a pCCI-Brick plasmid, resulting in plasmids entitled pHCA, pGPP, pCAN, pSCF, and pSOL as described in Table 3. See, FIGs. 15A-15E for the complete gene cassette inserted into the plasmids. Each of these plasmids then were used to amplify each synthetic gene cassette via standard plasmid prep. Plasmid DNA encoding each complete synthetic gene cassette was cloned into the Spel/Xhol cloning site of polycistronic yeast auxotrophic selection vectors, resulting in vectors entitled vHCA, vGPP, vCAN, vSCF, and vSOL as described in Table 3, to allow iterative antibiotic/auxotrophic selection of only those cells that were transformants of one or more such polycistronic vector(s).

TABLE 3

<td colspan="5"> HCA Gene Cassette</td>

<td> Gene ID</td><td> Cassette Position</td><td> pCCI-Brick #1 ID</td><td> Yeast Vector</td><td> Yeast Vector ID</td>

<td> ACL</td><td> 1</td><td rowspan="7"> pHCA</td><td rowspan="7"> pESC-TRP</td><td rowspan="7"> vHCA</td>

<td> atoB</td><td> 2</td>

<td> BHBD</td><td> 3</td>

<td> ECH</td><td> 4</td>

<td> ECR</td><td> 5</td>

<td> bktB</td><td> 6</td>

<td> HCS</td><td> 7</td>

<td colspan="5"> MVA Gene Cassette</td>

<td> Gene ID</td><td> Cassette Position</td><td> pCCI-Brick #2 ID</td><td> Yeast Vector</td><td> Yeast Vector ID</td>

<td> HMGS</td><td> 1</td><td rowspan="7"> pGPP</td><td rowspan="7"> pESC-LEU</td><td rowspan="7"> vGPP</td>

<td> tHMGR</td><td> 2</td>

<td> ERG12</td><td> 3</td>

<td> ERG8</td><td> 4</td>

<td> MVD1</td><td> 5</td>

<td> ID II</td><td> 6</td>

<td> ERG20^ww</td><td> 7</td>

<td colspan="5"> CAN Gene Cassette</td>

<td> Gene ID</td><td> Cassette Position</td><td> pCCI-Brick #3 ID</td><td> Yeast Vector</td><td> Yeast Vector ID</td>

<td> OS</td><td> 1</td><td rowspan="4"> pCAN</td><td rowspan="4"> pESC-HIS</td><td rowspan="4"> vCAN</td>

<td> OAC</td><td> 2</td>

<td> CBGAS</td><td> 3</td>

<td> CBDAS</td><td> 4</td>

<td> CBCAS</td><td> 5</td><td rowspan="2"></td><td rowspan="2"></td><td rowspan="2"></td>

<td> ACC</td><td> 6</td>

<td colspan="5"> SCFLD Gene Cassette</td>

<td> Gene ID</td><td> Cassette Position</td><td> pCCI-Brick #4 ID</td><td> Yeast Vector</td><td> Yeast Vector ID</td>

<td> CBSCF</td><td> 1</td><td rowspan="4"> pSCF</td><td rowspan="4"> pESC-URA #1</td><td rowspan="4"> vSCF</td>

<td> MCASCF</td><td> 2</td>

<td> ACL</td><td> 3</td>

<td> atoB</td><td> 4</td>

<td colspan="5"> NSCFLD Gene Cassette</td>

<td> Gene ID</td><td> Cassette Position</td><td> pCCI-Brick #5 ID</td><td> Yeast Vector</td><td> Yeast Vector ID</td>

<td> ACL</td><td> 1</td><td rowspan="2"> pSOL</td><td rowspan="2"> pESC-URA #2</td><td rowspan="2"> vSOL</td>

<td> atoB</td><td> 2</td>

The genes assigned to each synthetic gene cassette as well as the plasmids and vectors into which each synthetic gene cassette was inserted are listed in Table 3, the amino acid sequences encoded by each synthetic gene cassette are provided in FIGs.

13A-13D, the codon-optimized nucleotide sequence fragments comprising each synthetic gene cassette are detailed in FIGs. 14A-14D, the complete nucleotide sequences of each fully-assembled synthetic gene cassette (the complete insert sequences for each plasmid and expression vector) are provided in FIGs. 15A-15E, a general map of pCCI-Brick plasmids is shown in FIG. 16, and a general map of a polycistronic yeast auxotrophic selection vector is shown in FIG. 17.

Example 3 - Engineering of Cannabinoidergic Cells

To engineer a novel heterologous pathway for the biosynthesis of cannabinoids from citrate, and to evaluate the impacts of bidirectional multi-enzymatic scaffolding thereon, competent S. cerevisiae cells were sequent! ally/iteratively transformed with, and auxotrophically selected for, expression of vHCA, vGPP, vCAN, and either vSCF (for

scaffolded cannabinoid biosynthesis) or vSOL (for non-scaffolded/soluble cannabinoid biosynthesis) constructs.

All vector transformation and auxotrophic selection procedures were conducted as follows. An aliquot of an overnight S. cerevisiae culture was inoculated into 100mL YPD media (lOg/L yeast nitrogen base, 20g/L peptone, and 20g/L D-(+)-glucose) to OD600nm = 0.3 (stationary phase) and grown to OD600nm = 1.6 in an orbital shaker at 30°C and 225 RPM. Cells then were harvested by centrifugation at 3000 x g for 3 minutes followed by aspiration of media. The harvested cell pellet was next washed 2x with 50mL chilled nuclease-free water and lx with 50mL chilled electroporation buffer (IM sorbitol/lmM CaC12). Washed cells were conditioned by incubation for 30 minutes in 20mL 0.1M LiAc/lOmM DTT in an orbital shaker at 30°C and 225 RPM, harvested, washed lx with 50mL electroporation buffer, harvested, and resuspended in 100pL electroporation buffer. The resuspended cells were transformed with a quantity of vector containing 3 pg of the target DNA insert (calculated using the vector-insert ratio for each vector) by electroporation at 2.5kV and 25pF. To the electroporated cell suspension was then added 8mL of YPD media containing IM sorbitol, and the resulting suspension was incubated for one hour in an orbital shaker at 30°C and 225 RPM. To isolate target transformants by auxotrophic selection, cells were harvested, resuspended in the appropriate yeast nitrogen base (YNB) dropout (selection) media as subsequently described for each iterative transformation step, transferred to a baffled culture flask, and incubated overnight in an orbital shaker at 30°C and 225 RPM. The transformation and selection protocols were utilized sequentially for each assigned vector.

Applying the aforementioned approach, an initial culture of electrocompetent S. cerevisiae cells was first transformed with vHCA, which encodes scaffold-binding engineered enzymes required for biosynthesis of HCA from citrate. Cells transformed with vHCA (designated yHCA) were selected for by resuspension and incubation in tryptophan-deficient YNB media. Selected yHCA cells (i.e., cells that grew in tryptophan-deficient YNB media) were next transformed with vGPP, which encodes scaffold-binding engineered enzymes required for biosynthesis of GPP from citrate. Cells co-transformed with vHCA and vGPP (designated yHCAGPP) were selected for by

resuspension and incubation in tryptophan- and leucine-deficient YNB media. Selected yHCAGPP cells (i.e., cells that grew in tryptophan- and leucine-deficient YNB media) were then transformed with vCAN, which encodes scaffold-binding engineered enzymes required for biosynthesis of malonyl-CoA from citrate, olivetol from HCA and malonylC0A, OVA (olivetoic acid) from olivetol, and CBGA from OVA and GPP as well as soluble enzymes required for biosynthesis of CBDA and CBCA from CBGA). Cells cotransformed with vHCA, vGPP, and vCAN (designated yCBparent) were selected for by resuspension and incubation in tryptophan-, leucine-, and histidine-deficient YNB media.

The yCBparent culture containing cells that grew in tryptophan-, leucine-, and histidine-deficient YNB media then was split into two separate cultures. The first of the split yCBparent cultures was transformed with vSCF, which encodes CBSCF (cannabinoidergic metabolon scaffold) and MCASCF (malonyl-CoA metabolon scaffold) as well as additional copies of ACL and atoB. Cells co-transformed with vHCA, vGPP, vCAN, and vSCF (designated yCBscr) were selected for by resuspension and incubation in tryptophan-, leucine-, histidine-, and uracil-deficient YNB media. The second of the split yCBparent cultures was transformed with vSOL, which encodes additional copies of ACL and atoB but lacks both CBSCF and MCASCF. Cells co-transformed with vHCA, vGPP, vCAN, and vSOL (designated yCBsor) were also selected for by resuspension and incubation in tryptophan-, leucine-, histidine-, and uracil-deficient YNB media.

To quantify the improvement in cannabinoidergic capacity conferred by multienzymatic scaffolding, cannabinoid titers were compared between triplicate yCBsoL and yCBscr cultures grown in 100mL YPD media for 48 hours at 30°C and 400 RPM in an incubator-shaker. To compare the proliferation rates of yCBsoL and yCBscr, each culture was initially diluted to OD600nm = 0.3, and OD600nm measurements were recorded in 12hour intervals thereafter. Proliferation curves are depicted in FIG. 18. The extra sum-ofsquares F-test indicated that the proliferation curves of yCBSCF and yCBSOL cultures did not significantly differ for any parameter over the 48-hour incubation period, indicating that scaffolding does not impact cellular proliferation.

Total cannabinoid titers, parent (carboxylated) cannabinoid (CBGA, CBDA, and CBCA) titers, derivative (decarboxylated) cannabinoid (CBG, CBD, and CBC) titers, and

cannabinoid precursor (OVA) titers were measured. As shown in FIG. 19, mixed ANOVA detected main effects of strain (F1,4=943.8; p&lt;0.0001) and analyte (cannabinoid and cannabinoid precursor) titers (F10,40=216.4; p&lt;0.0001) and a significant strain x analyte interaction (F10,40= 131.4; p&lt;0.0001). Relative to yCBSOL cultures, yCBSCF cultures exhibited increased total cannabinoid (p&lt;0.0001), OVA precursor (p&lt;0.0001), CBG(A) (p&lt;0.0001), CBD(A) (p&lt;0.0001), CBC(A) (p&lt;0.0001), CBGA (p&lt;0.0001), CBDA (p&lt;0.0001), CBCA (p&lt;0.0001), CBG (p&lt;0.0001), CBD (p&lt;0.01), and CBC (p&lt;0.001) titers.

Example 4 - Impacts of Citrate and Hexanoate Supplementation on Scaffolded and Soluble Cannabinoid Biosynthesis

To evaluate the impacts of culture media supplementation with citrate and hexanoate precursors, cannabinoid titers were compared between triplicate yCBsoL and yCBscF cultures grown in 100mL YPD media containing 300 mg/L of either buffered citrate (pH 6.0) or hexanoate for 48 hours at 30°C and 400 RPM in an orbital shaker. All cultures were initially diluted to OD600nm = 0.3. Cannabinoid titers for cultures grown in YPD media, citrate-supplemented YPD media, and hexanoate-supplemented YPD media were assessed and analyzed by ANOVA. As shown in FIG. 20, mixed ANOVA detected main effects of strain (F1,4=457.5; p&lt;0.0001) and culture media supplementation (F2,8=312.5; p&lt;0.0001) and a significant strain x culture media supplementation interaction (F2,8=289.6; p&lt;0.0001). Compared to basal media cultures, yCBSCF but not yCBSOL cultures exhibited increased total cannabinoid titers when cultured in media supplemented with 300mg/L citrate (p&lt;0.0001). Neither yCBSCF nor yCBSOL cultures differed in total cannabinoid titers relative to basal media when cultured in media supplemented with 300mg/L hexanoate. For all measures, n = 3 biological replicates for yCBSCF and yCBSOL cultures. Moreover, relative to yCBSOL cultures, yCBSCF cultures exhibited increased total cannabinoid titers when cultured in basal media (p&lt;0.0001, data also reported in Figure 19) as well as media supplemented with 300mg/L citrate (p&lt;0.0001) and hexanoate (p&lt;0.0001).

To delineate concentration-response relationships for the supplementation of culture media with citrate, cannabinoid titers were compared between triplicate yCBsoL and yCBscF cultures grown in 100mL YPD media containing 0, 10, 30, 100, 300, 1000, 3000, and 10000 mg/L buffered citrate (pH 6.0) for 48 hours at 30°C and 400 RPM in an orbital shaker. All cultures were initially diluted to OD600nm = 0.3. Following quantification, asymmetric sigmoidal (five-parameter) logistic regressions were computed to fit concentration-response curves, from which were derived estimates of the maximal cannabinoid titer (CBMax) and citrate EC50 for cannabinoid biosynthesis in yCBsoL and yCBscr cultures. Concentration-response curves, CBMax estimates, and citrate EC50 estimates are depicted in Figure 21. Mixed ANOVA detected main effects of strain (F1,8=69.9; p&lt;0.0001) and parameter (F1,8=66.7; p&lt;0.0001) and a significant strain x parameter interaction (F1,8=5.3; p&lt;0.05) for concentration-response parameter estimates (CBMax and citrate EC50). Compared to yCBSOL cultures, yCBSCF cultures exhibited markedly increased CBMax (p&lt;0.0001) and citrate EC50 (p&lt;0.001) estimates.

OTHER EMBODIMENTS

It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

<img file="IL283384A_D0001.tif" />

<img file="IL283384A_D0002.tif" />

<img file="IL283384A_D0003.tif" />

<img file="IL283384A_D0004.tif" />

<img file="IL283384A_D0005.tif" />

Malonyl-CoA Pathway

<img file="IL283384A_D0006.tif" />

Acetoacetyl 3-Hydroxy- Trans-But Butanoyl-C0A Butanoyl-CoA enoyl-CoA C0A

<img file="IL283384A_D0007.tif" />

<img file="IL283384A_D0008.tif" />

Hexanoyl-CoA Pathway

<img file="IL283384A_D0009.tif" />

<img file="IL283384A_D0010.tif" />

FIG. 1A (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0011.tif" />

.ω ω co

3,5,7trioxododecanoylC0A

<img file="IL283384A_D0012.tif" />

MalonylCitrate Acetyl-CoA

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0013.tif" />

ο ο

&amp;#1502; &amp;#1504;&amp;#1505;

JD1Z

&amp;#1504;&amp;#1505;

ID1

HexanoylC0A

Olivetol Synthase (OS)

<img file="IL283384A_D0014.tif" />

&amp;#1504;&amp;#1505;

JD14

ID17 Scaffolded Ligands

ID1 Scaffolded Ligands μι μι &lt;λ

Τ rans-Hex Hydroxy3-ketoHexanoyl-CoA Hexanoyl-CoA enoyl-CoA

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

ID6 Scaffolded

ID3 Scaffolded Ligands

ID4 Scaffolded Ligands

ID5

ID14

<td> Scaffolded</td><td></td><td> Scaffolded</td>

Ligands

Ligands

FIG. 1A (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0015.tif" />

<img file="IL283384A_D0016.tif" />

CH, CH, OH O

<img file="IL283384A_D0017.tif" />

Cannabigerolic Acid

HoC 0 HO

OH

CH3

H3C H

CH

J, R

<img file="IL283384A_D0018.tif" />

H

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0019.tif" />

HO

Cannabigerolic Acid

CH3

H2Q

<img file="IL283384A_D0020.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid

OH

CH3 CH3 OH Ο

<img file="IL283384A_D0021.tif" />

HO

CBDA Synthase ^(CBDAS)^

<img file="IL283384A_D0022.tif" />

Cannabigerolic Acid

H31

CH3

<img file="IL283384A_D0023.tif" />

IPP

MPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

R

S

Olivetol Acid Cyclase . (OAC) , o

&amp;#1504;&amp;#1505;

ID15

<img file="IL283384A_D0024.tif" />

ID15 Scaffolded Ligands

CBGA Synthase (CBGAS) o

&amp;#1504;&amp;#1505;

ID16

<img file="IL283384A_D0025.tif" />

ID16 Scaffolded Ligands

Upper Cannabinoid Pathway

GPP Synthase (ERG20ww) F96W\N127W/ o

&amp;#1504;&amp;#1505;

ID13

<img file="IL283384A_D0026.tif" />

ID13 Scaffolded Ligands

Isopentyl Diphosphate Isomerase

Diphosphomevalonate Decarboxylase \(MVD1)/

&amp;#1504;&amp;#1505;

JD12

&amp;#1504;&amp;#1505;

ID11

ID12 Scaffolded Ligands

ID11 Scaffolded Ligands

FIG. 1A (Continuous)

SUBSTITUTE SHEET (RULE 26) &lt;U) O &lt;R

Δ hv

Δ ------> hv

3U>=^CH3 H3C Q°

H0XV ch3

Cannabichromene

<img file="IL283384A_D0027.tif" />

CH, OH h2c

H3C HO ch Cannabidiol

<img file="IL283384A_D0028.tif" />

R

S

LU

<img file="IL283384A_D0029.tif" />

Malonyl-CoA Pathway

Upper CB Pathway + CBCAS + CBDAS

<img file="IL283384A_D0030.tif" />

CBergic + MCA Pathway Scaffolds 13

<img file="IL283384A_D0031.tif" />

Hexanoyl-CoA Pathway

Mevalonate Pathway

Nucleus

<img file="IL283384A_D0032.tif" />

CH3 CHoOHO h3c

Cannabigerolic Acid ch3

<img file="IL283384A_D0033.tif" />

Mevalonate , ± *

phosphate Mevalonate HMG-C0A

Cytosol hv ch3 ch3

<img file="IL283384A_D0034.tif" />

Cannabigerol H&amp;#1470;C HO

Acetoacetyl- AcetylC0A C0A

Phospho Mevalonate Truncated HMG-C0A Acetyl-CoA

Mevalonate

&amp;#1504;&amp;#1505;

JD1Q

ID10

Kinase

&amp;#1504;&amp;#1505;

ID9

ID9

HMG-C0A Synthase

&amp;#1504;&amp;#1505;

ID8

ID8

Acetylch3 o o ω

<img file="IL283384A_D0035.tif" />

Citrate

ATP Citrate Lyase

&amp;#1504;&amp;#1505;

ID7

ID7

&amp;#1504;&amp;#1505;

ID2

ID2

&amp;#1504;&amp;#1505;

ID1

ID1

Scaffolded _ Scaffolded _ Scaffolded Scaffolded Scaffolded Scaffolded J

Ligands

Ligands

Ligands

Ligands

Ligands

Ligands

<img file="IL283384A_D0036.tif" />

X

Mevalonate Pathway

FIG. 1A

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0037.tif" />

<img file="IL283384A_D0038.tif" />

<img file="IL283384A_D0039.tif" />

<img file="IL283384A_D0040.tif" />

HO Ο

<img file="IL283384A_D0041.tif" />

Malonyl-CoA Pathway

<img file="IL283384A_D0042.tif" />

Acetoacetyl 3-Hydroxy- Trans-But C0A Butanoyl-CoA enoyl-CoA

<img file="IL283384A_D0043.tif" />

<img file="IL283384A_D0044.tif" />

ButanoylC0A

<img file="IL283384A_D0045.tif" />

Hexanoyl-CoA Pathway

FIG. 1B (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0046.tif" />

<img file="IL283384A_D0047.tif" />

<img file="IL283384A_D0048.tif" />

<img file="IL283384A_D0049.tif" />

<img file="IL283384A_D0050.tif" />

3,5,7trioxododecanoylC0A

<img file="IL283384A_D0051.tif" />

Olivetol Synthase (OS)

<img file="IL283384A_D0052.tif" />

&amp;#1504;&amp;#1505;

JD14

ID14 Scaffolded Ligands

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

ID5

<td> Scaffolded</td><td></td>

Ligands

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

ID4 Scaffolded Ligands

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

ID3 Scaffolded Ligands

FIG. 1B (Continuous)

SUBSTITUTE SHEET (RULE 26)

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

ID6 Scaffolded

<img file="IL283384A_D0053.tif" />

<img file="IL283384A_D0054.tif" />

<img file="IL283384A_D0055.tif" />

CH, CH, OH O

<img file="IL283384A_D0056.tif" />

Cannabigerolic Acid

HoC 0 HO

<img file="IL283384A_D0057.tif" />

H h3&amp;#1523;

OH

CH3

H3C H

CH

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0058.tif" />

HO

Cannabigerolic Acid

OH

CH3 CH3 OH O

<img file="IL283384A_D0059.tif" />

HO ch3

CH3

H2Q

<img file="IL283384A_D0060.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid

CBDA Synthase ^(CBDAS)^

<img file="IL283384A_D0061.tif" />

Cannabigerolic Acid

<img file="IL283384A_D0062.tif" />

IPP

MPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

R

S

Olivetol Acid Cyclase . (OAC) , o

&amp;#1504;&amp;#1505;

ID15

<img file="IL283384A_D0063.tif" />

ID15 Scaffolded Ligands

CBGA Synthase (CBGAS) o

&amp;#1504;&amp;#1505;

ID16

<img file="IL283384A_D0064.tif" />

ID16 Scaffolded Ligands

Upper Cannabinoid Pathway

GPP Synthase (ERG20ww) F96W\N127W/ o

&amp;#1504;&amp;#1505;

ID13

<img file="IL283384A_D0065.tif" />

ID13 Scaffolded Ligands

Isopentyl Diphosphate Isomerase

DiphosphoMevalonate Decarboxylase \(MVD1)/

&amp;#1504;&amp;#1505;

JD12

&amp;#1504;&amp;#1505;

ID11

ID12 Scaffolded Ligands

ID11 Scaffolded Ligands

FIG. 1B (Continuous)

SUBSTITUTE SHEET (RULE 26) &lt;U) O &lt;R

8/156 hv hv H3C>vCH3 H3C Q?

H0XV ch3

Cannabichromene

CH^

OH

<img file="IL283384A_D0066.tif" />

h2q h3c HO ch.

Cannabidiol 13

<img file="IL283384A_D0067.tif" />

GPP Cassette

CAN Cassette

SCF Cassette

Cytosol

HCA Cassette

Nucleus

<img file="IL283384A_D0068.tif" />

<img file="IL283384A_D0069.tif" />

H3C

R

S CH3 CHoOHO

I I &amp;#1506;&amp;#1497; I II ch3 ch3

Cannabigerolic Acid ch3 hv

<img file="IL283384A_D0070.tif" />

Cannabigerol H&amp;#1470;C HO ch3

<img file="IL283384A_D0071.tif" />

Mevalonate , ± *

phosphate Mevalonate HMG-C0A

Acetoacetyl- AcetylC0A C0A

<img file="IL283384A_D0072.tif" />

Citrate ω ω

Phospho Mevalonate Truncated HMG-C0A Acetyl-CoA

Mevalonate

&amp;#1504;&amp;#1505;

JD1Q

ID10

Kinase

&amp;#1504;&amp;#1505;

ID9

ID9

HMG-C0A Synthase

&amp;#1504;&amp;#1505;

ID8

ID8

AcetylATP Citrate

Lyase

&amp;#1504;&amp;#1505;

ID7

ID7

&amp;#1504;&amp;#1505;

ID2

ID2

&amp;#1504;&amp;#1505;

ID1

ID1

Scaffolded _ Scaffolded _ Scaffolded Scaffolded Scaffolded Scaffolded J

Ligands

Ligands

Ligands

Ligands

Ligands

Ligands

<img file="IL283384A_D0073.tif" />

X

Mevalonate Pathway

FIG. 1B

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0074.tif" />

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0075.tif" />

igure 2A (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0076.tif" />

<img file="IL283384A_D0077.tif" />

<img file="IL283384A_D0078.tif" />

<img file="IL283384A_D0079.tif" />

<img file="IL283384A_D0080.tif" />

<img file="IL283384A_D0081.tif" />

<img file="IL283384A_D0082.tif" />

<img file="IL283384A_D0083.tif" />

igure 2A (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0084.tif" />

igure 2A

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0085.tif" />

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0086.tif" />

igure 2B (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0087.tif" />

igure 2B (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0088.tif" />

igure 2B

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0089.tif" />

<img file="IL283384A_D0090.tif" />

<img file="IL283384A_D0091.tif" />

Ligand #2 Linker Ligand #1 (Scaffolded ID-binding Domain)

FIG. 3

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0092.tif" />

Di hydroAcetyl-CoA

Pyruvate Dehydrogenase Lipoyl-trans-

<img file="IL283384A_D0093.tif" />

DihydroLipoyl-trans- Carboxylase (ACC) acetylase o

Pyruvate Dehydrogenase \ (E1)

&amp;#1502; &amp;#1504;&amp;#1505;

JDX

&amp;#1502; &amp;#1504;&amp;#1505;

JDX

&amp;#1504;&amp;#1505;

JDX

&amp;#1504;&amp;#1505;

JDX

&amp;#1504;&amp;#1505;

JDX

IDX

IDX

IDX

IDX

Q Scaffolded Scaffolded__ Ligands |

Ligands |2 Scaffolded _ Scaffolded _ Scaffolded &amp;#1498;

Ligands

Ligands

Ligands

Malonyl-CoA Pathway

<img file="IL283384A_D0094.tif" />

<img file="IL283384A_D0095.tif" />

I

<img file="IL283384A_D0096.tif" />

<img file="IL283384A_D0097.tif" />

Pyruvate

2-Hydroxyethyl-TPP

Acetyl- Acetoacetyl 3-Hydroxy- Trans-But-2 Butanoyl

C0A -C0A Butanoyl-CoA -enoyl-CoA -C0A

<img file="IL283384A_D0098.tif" />

T rans-2enoyl-CoA-

&amp;#1504;&amp;#1505;

ID5

ID5 Scaffolded

Enoyl-CoA Hydratase

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

ID4 Scaffolded Ligands

3-hydroxybutyryl-CoA De-

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

ID3

Acetyl-CoA AcetylT ransferase

DihydroLipoyl-trans-

&amp;#1504;&amp;#1505;

ID2

ID2

Pyruvate Dehydrogenase

&amp;#1502; &amp;#1504;&amp;#1505;

JDX

IDX

&amp;#1504;&amp;#1505;

JDX.

IDX

[ Scaffolded Scaffolded Scaffolded Scaffolded

Ligands

Ligands

Ligands

Hexanoyl-CoA Pathway

Ligands

<img file="IL283384A_D0099.tif" />

FIG. 4 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0100.tif" />

Acetyl-CoA

Pyruvate

MalonylC0A

2-Hydroxyethyl-TPP

MalonylC0A

Acetyl-CoA Carboxylase &lt; (ACC) J

Dihydrolipoyl-transacetylase \(E2)/

O

CO

Pyruvate Dehydrogenase X (E1) X

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0101.tif" />

o

&amp;#1502; &amp;#1504;&amp;#1505;

JDX

<img file="IL283384A_D0102.tif" />

o .ω

3,5,7trioxododecanoylC0A

&amp;#1504;&amp;#1505;

JDX

&amp;#1504;&amp;#1505;

JDX

&amp;#1504;&amp;#1505;

JDX

IDX

IDX

IDX

Q Scaffolded _ Scaffolded _ Scaffolded &amp;#1498;

Ligands

Ligands

Ligands

Ii IDX Scaffolded &amp;#1498;

Γ Ligands hexanoylC0A

Olivetol Synthase (OS)

<img file="IL283384A_D0103.tif" />

&amp;#1504;&amp;#1505;

JD14

ID14 Scaffolded Ligands !4) co

T rans-Hex HydroxyHexanoyl-CoA enoyl-CoA

3-ketoHexanoyl-CoA

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

ID5

<td> Scaffolded</td><td></td>

Ligands

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

ID4 Scaffolded Ligands

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)X

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

ID3 Scaffolded Ligands

FIG. 4 (Continuous)

SUBSTITUTE SHEET (RULE 26)

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

ID6 Scaffolded

L

<img file="IL283384A_D0104.tif" />

<img file="IL283384A_D0105.tif" />

h3&amp;#1523;

CH, CH, OH O

<img file="IL283384A_D0106.tif" />

HO

Cannabigerolic Acid

OH

CH3

H3C H

CH

<img file="IL283384A_D0107.tif" />

H

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0108.tif" />

HO

Cannabigerolic Acid ch3 ch3 oh o

ΗοΟ'^^Ο&amp;#972;&amp;#972; 3 HO^^

OH ch3

CH3 h2Q

<img file="IL283384A_D0109.tif" />

<img file="IL283384A_D0110.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid

CBDA Synthase ^(CBDAS)^

Cannabigerolic Acid

<img file="IL283384A_D0111.tif" />

IPP

MPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

R

S

Olivetol Acid Cyclase . (OAC) ,

&amp;#1504;&amp;#1505;

JD15

ID15 Scaffolded Ligands

CBGA Synthase (CBGAS)

&amp;#1504;&amp;#1505;

JD16

ID16 Scaffolded Ligands

Upper Cannabinoid Pathway

GPP Synthase (ERG20ww) F96W\N127W/

Isopentyl Diphosphate Isomerase \(IDI1)/

Diphosphomevalonate Decarboxylase \(MVD1)/

&amp;#1504;&amp;#1505;

JD13

&amp;#1504;&amp;#1505;

JD12

&amp;#1504;&amp;#1505;

ID11

ID13 Scaffolded Ligands

ID12 Scaffolded Ligands

ID11 Scaffolded Ligands &lt;U) O &lt;R

FIG. 4 (Continuous)

SUBSTITUTE SHEET (RULE 26)

Upper CB Pathway + CBCAS + CBDAS

<img file="IL283384A_D0112.tif" />

<img file="IL283384A_D0113.tif" />

Malonyl-CoA Pathway ch3

3Y=\CH3 H3C Q°

H0XV

Cannabichromene ch3

CBergic + MCA Pathway Scaffolds

Mevalonate Pathway

Nucleus

<img file="IL283384A_D0114.tif" />

<img file="IL283384A_D0115.tif" />

Hexanoyl-CoA Pathway

Cytosol

CH3 CM H&amp;#1470;C HO

<img file="IL283384A_D0116.tif" />

Cannabigerol hv ch3

Pyruvate

2-hydrooxyethyl -TPP

Acetoacetyl- AcetylC0A

C0A

<img file="IL283384A_D0117.tif" />

CH, OH

H2C h3c ho CH

Cannabidiol CH3 CH, OHO h3c

Cannabidiolic Acid

<img file="IL283384A_D0118.tif" />

Δ hv hv

<img file="IL283384A_D0119.tif" />

R

S

Mevalonate. a

phosphate Mevalonate HMG-C0A

Mevalona-||Truncated||HMG-CoA|| Acetyl-CoA Dihydro- Pyruvate

Lipoyl-tran- DehydroPhosphomevalonat Kinase \(ERG8)/

HMG-C0A y_________ AcetylReductase l/HMGsJh'ransferasellsacetylaseJI genase

Synthase ;tHMGR).

te Kinase (ERG12)

&amp;#1504;&amp;#1505;

IDX

&amp;#1504;&amp;#1505;

IDX

&amp;#1504;&amp;#1505;

&amp;#1504;&amp;#1505;

&amp;#1504;&amp;#1505;

&amp;#1504;&amp;#1505;

IDX ScIDX Scaffolded _ affolded ] Ligands J ^Ligands

ID10 Sc-H ID9 Sc- |_&amp;#943; ID8 Sc- |_&amp;#943; ID7 Sc- |_i ID2 Sc-

<img file="IL283384A_D0120.tif" />

M affolded H affolded H affolded H affolded N affolded 1 LigandsJ I LigandsJ I LigandsJ I LigandsJ I Ligands

Ligands

Ligands

Ligands

Ligands

Mevalonate Pathway

FIG. 4

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0121.tif" />

<img file="IL283384A_D0122.tif" />

<img file="IL283384A_D0123.tif" />

<img file="IL283384A_D0124.tif" />

<img file="IL283384A_D0125.tif" />

Malonyl-CoA Pathway

<img file="IL283384A_D0126.tif" />

Acetoacetyl 3-Hydroxy- Trans-But C0A Butanoyl-CoA enoyl-CoA

<img file="IL283384A_D0127.tif" />

<img file="IL283384A_D0128.tif" />

ButanoylC0A

<img file="IL283384A_D0129.tif" />

Hexanoyl-CoA Pathway

<img file="IL283384A_D0130.tif" />

<img file="IL283384A_D0131.tif" />

FIG. 5 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0132.tif" />

.ω ω co

3,5,7trioxododecanoylC0A

<img file="IL283384A_D0133.tif" />

MalonylCitrate Acetyl-CoA

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0134.tif" />

ο ο

&amp;#1502; &amp;#1504;&amp;#1505;

ID1

&amp;#1502; &amp;#1504;&amp;#1505;

JD1Z

HexanoylC0A

Olivetol Synthase (OS)

<img file="IL283384A_D0135.tif" />

&amp;#1504;&amp;#1505;

JD14

ID14 Scaffolded Ligands

ID17 Scaffolded Ligands

ID1 Scaffolded Ligands (Λ

Τ rans-Hex Hydroxy3-ketoHexanoyl-CoA Hexanoyl-CoA enoyl-CoA

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

ID5

<td> Scaffolded</td><td></td>

Ligands

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

ID4 Scaffolded Ligands

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

ID3 Scaffolded Ligands

FIG. 5 (Continuous)

SUBSTITUTE SHEET (RULE 26)

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

ID6 Scaffolded

L

<img file="IL283384A_D0136.tif" />

<img file="IL283384A_D0137.tif" />

CH, CH, OH O

<img file="IL283384A_D0138.tif" />

Cannabigerolic Acid

HoC 0 HO

<img file="IL283384A_D0139.tif" />

H h3'

OH

CH3

H3C H

CH

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0140.tif" />

HO

Cannabigerolic Acid ch3

H2Q

<img file="IL283384A_D0141.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid

OH ch3 ch3 oh o

<img file="IL283384A_D0142.tif" />

HO ch3

CBDA Synthase ^fCBDAS)^

<img file="IL283384A_D0143.tif" />

Cannabigerolic Acid

<img file="IL283384A_D0144.tif" />

cMEPP

Olivetolic Acid

R

S

Geranyl

Pyrophosphate IPP + DMAPP HMBPP

CBGA Synthase (CBGAS)

Olivetol Acid Cyclase (OAC)

<img file="IL283384A_D0145.tif" />

o o

GPP Synthase (ERG20ww) F96W\N127W/

HMBPP Reductase . (ISPH) /

HMBPP Synthase (ISPG)

&amp;#1504;&amp;#1505;

ID15

<img file="IL283384A_D0146.tif" />

ID15 Scaffolded Ligands

&amp;#1504;&amp;#1505;

ID16

<img file="IL283384A_D0147.tif" />

ID16 Scaffolded Ligands o

&amp;#1504;&amp;#1505;

IDX

<img file="IL283384A_D0148.tif" />

IDX Scaffolded Ligands

&amp;#1504;&amp;#1505;

IDX

<img file="IL283384A_D0149.tif" />

IDX Scaffolded Ligands o

&amp;#1504;&amp;#1505;

IDX

<img file="IL283384A_D0150.tif" />

IDX Scaffolded Ligands

Upper Cannabinoid Pathway

FIG. 5 (Continuous)

SUBSTITUTE SHEET (RULE 26) &lt;%)

<img file="IL283384A_D0151.tif" />

Cannabichromene

<img file="IL283384A_D0152.tif" />

Malonyl-CoA Pathway

Upper CB Pathway + CBCAS + CBDAS

<img file="IL283384A_D0153.tif" />

hv

CH, OH h2c h3c HO ch Cannabidiol

<img file="IL283384A_D0154.tif" />

CBergic + MCA Pathway Scaffolds CH3 CHoOHO 13

<img file="IL283384A_D0155.tif" />

MEP Pathway

Hexanoyl-CoA Pathway

Nucleus

<img file="IL283384A_D0156.tif" />

Cytosol

<img file="IL283384A_D0157.tif" />

R

S ch3

CH3 CH H&amp;#1470;C HO

<img file="IL283384A_D0158.tif" />

Cannabigerolic Acid [ hv

Cannabigerol

CH3

<img file="IL283384A_D0159.tif" />

Pyruvate + G3P

CDP-MEP

CDP-ME

DOXP

MEP

MECDP

Synthase

&amp;#1504;&amp;#1505;

IDX

IDX Scaffolded Ligands

CDPME Kinase (ISPE)

MEP Cytidyl T ransferase

DOXP Reductoisomerase

DOXP Synthase

&amp;#1504;&amp;#1505;

IDX

&amp;#1504;&amp;#1505;

JDX

&amp;#1504;&amp;#1505;

IDX

&amp;#1504;&amp;#1505;

IDX

IDX Scaffolded Ligands

IDX Scaffolded Ligands

IDX Scaffolded Ligands

IDX Scaffolded Ligands

<img file="IL283384A_D0160.tif" />

FIG. 5

X

MEP Pathway

SUBSTITUTE SHEET (RULE 26)

Figure 6A

ATP Citrate Lyase

MSAKAISEQTGKELLYKFICTTSAIQNRFKYARVTPDTDWARLLQDHPWLLSQNLWKPDQLIKRRGKLGLVGVNLTL DGVKSWLKPRLGQEATVGKATGFLKNFLIEPFVPHSQAEEFYVCIYATREGDYVLFHHEGGVDVGDVDAKAQKLLV GVDEKLNPEDIKKHLLVHAPEDKKEILASFISGLFNFYEDLYFTYLEINPLWTKDGVYVLDLAAKVDATADYICKVKWG DIEFPPPFGREAYPEEAYIADLDAKSGASLKLTLLNPKGRIWTMVAGGGASWYSDTICDLGGVNELANYGEYSGAP SEQQTYDYAKTILSLMTREKHPDGKILIIGGSIANFTNVAATFKGIVRAIRDYQGPLKEHEVTIFVRRGGPNYQEGLRV MGEVGKTTGIPIHVFGTETHMTAIVGMALGHRPIPNQPPTAAHTANFLLNASGSTSTPAPSRTASFSESRADEVAPAK KAKPAMPQDSVPSPRSLQGKSTTLFSRHTKAIVWGMQTRAVQGMLDFDYVCSRDEPSVAAMVYPFTGDHKQKFY WGHKEILIPVFKNMADAMRKHPEVDVLINFASLRSAYDSTMETMNYAQIRTIAIIAEGIPEALTRKLIKKADQKGVTIIGP ATVGGIKPGCFKIGNTGGMLDNILASKLYRPGSVAYVSRSGGMSNELNNIISRTTDGVYEGVAIGGDRYPGSTFMDH VLRYQDTPGVKMIWLGEIGGTEEYKICRGIKEGRLTKPIVCWCIGTCATMFSSEVQFGHAGACANQASETAVAKNQ ALKEAGVFVPRSFDELGEIIQSVYEDLVANGVIVPAQEVPPPTVPMDYSWARELGLIRKPASFMTSICDERGQELIYA GMPITEVFKEEMGIGGVLGLLWFQKRLPKYSCQFIEMCLMVTADHGPAVSGAHNTIICARAGKDLVSSLTSGLLTIGD RFGGALDAAAKMFSKAFDSGIIPMEFVNKMKKEGKLIMGIGHRVKSINNPDMRVQILKDYVRQHFPATPLLDYALEVE KITTSKKPNLILNVDGLIGVAFVDMLRNCGSFTREEADEYIDIGALNGIFVLGRSMGFIGHYLDQKRLKQGLYRHPWD DISYVLPEHMSM

Acetyl-CoA Acetyltransferase (atoB)

MKNCVIVSAVRTAIGSFNGSLASTSAIDLGATVIKAAIERAKIDSQHVDEVIMGNVLQAGLGQNPARQALLKSGLAETV CGFTVNKVCGSGLKSVALAAQAIQAGQAQSIVAGGMENMSLAPYLLDAKARSGYRLGDGQVYDVILRDGLMCATH GYHMGITAENVAKEYGITREMQDELALHSQRKAAAAIESGAFTAEIVPVNWTRKKTFVFSQDEFPKANSTAEALGAL RPAFDKAGTVTAGNASGINDGAAALVIMEESAALAAGLTPLARIKSYASGGVPPALMGMGPVPATQKALQLAGLQLA DIDLIEANEAFAAQFLAVGKNLGFDSEKVNVNGGAIALGHPIGASGARILVTLLHAMQARDKTLGLATLCIGGGQGIAM VIERLN

3-Hvdroxvbutvryl-CoA Dehydrogenase

MKKVCVIGAGTMGSGIAQAFAAKGFEWLRDIKDEFVDRGLDFINKNLSKLVKKGKIEEATKVEILTRISGTVDLNMAA DCDLVIEAAVERMDIKKQIFADLDNICKPETILASNTSSLSITEVASATKRPDKVIGMHFFNPAPVMKLVEVIRGIATSQE TFDAVKETSIAIGKDPVEVAEAPGFWNRILIPMINEAVGILAEGIASVEDIDKAMKLGANHPMGPLELGDFIGLDICLAI MDVLYSETGDSKYRPHTLLKKYVRAGWLGRKSGKGFYDYSK

Enoyl-CoA Hydratase

MELNNVILEKEGKVAWTINRPKALNALNSDTLKEMDYVIGEIENDSEVLAVILTGAGEKSFVAGADISEMKEMNTIEG RKFGILGNKVFRRLELLEKPVIAAVNGFALGGGCEIAMSCDIRIASSNARFGQPEVGLGITPGFGGTQRLSRLVGMGM AKQLIFTAQNIKADEALRIGLVNKVVEPSELMNTAKEIANKIVSNAPVAVKLSKQAINRGMQCDIDTALAFESEAFGECF STEDQKDAMTAFIEKRKIEGFKNR

Trans-Enoyl-CoA Reductase

MIVKPMVRNNICLNAHPQGCKKGVEDQIEYTKKRITAEVKAGAKAPKNVLVLGCSNGYGLASRITAAFGYGAATIGVS FEKAGSETKYGTPGWYNNLAFDEAAKREGLYSVTIDGDAFSDEIKAQVIEEAKKKGIKFDLIVYSLASPVRTDPDTGIM HKSVLKPFGKTFTGKTVDPFTGELKEISAEPANDEEAAATVKVMGGEDWERWIKQLSKEGLLEEGCITLAYSYIGPEA TQALYRKGTIGKAKEHLEATAHRLNKENPSIRAFVSVNKGLVTRASAVIPVIPLYLASLFKVMKEKGNHEGCIEQITRLY AERLYRKDGTIPVDEENRIRIDDWELEEDVQKAVSALMEKVTGENAESLTDLAGYRHDFLASNGFDVEGINYEAEVE RFDRI

Figure 6A (continued)

Beta-Ketothiolase (bktB)

MTREVWVSGVRTAIGTFGGSLKDVAPAELGALWREALARAQVSGDDVGHWFGNVIQTEPRDMYLGRVAAVNG GVTINAPALTVNRLCGSGLQAIVSAAQTILLGDTDVAIGGGAESMSRAPYLAPAARWGARMGDAGLVDMMLGALHD PFHRIHMGVTAENVAKEYDISRAQQDEAALESHRRASAAIKAGYFKDQIVPWSKGRKGDVTFDTDEHVRHDATIDD MTKLRPVFVKENGTVTAGNASGLNDAAAAVVMMERAEAERRGLKPLARLVSYGHAGVDPKAMGIGPVPATKIALER AGLQVSDLDVIEANEAFAAQACAVTKALGLDPAKVNPNGSGISLGHPIGATGALITVKALHELNRVQGRYALVTMCIG GGQGIAAIFERI

HMG-CoA Synthase

MKLSTKLCWCGIKGRLRPQKQQQLHNTNLQMTELKKQKTAEQKTRPQNVGIKGIQIYIPTQCVNQSELEKFDGVSQ GKYTIGLGQTNMSFVNDREDIYSMSLTVLSKLIKSYNIDTNKIGRLEVGTETLIDKSKSVKSVLMQLFGENTDVEGIDTL NACYGGTNALFNSLNWIESNAWDGRDAIWCGDIAIYDKGAARPTGGAGTVAMWIGPDAPIVFDSVRASYMEHAYD FYKPDFTSEYPYVDGHFSLTCYVKALDQVYKSYSKKAISKGLVSDPAGSDALNVLKYFDYNVFHVPTCKLVTKSYGR LLYNDFRANPQLFPEVDAELATRDYDESLTDKNIEKTFVNVAKPFHKERVAQSLIVPTNTGNMYTASVYAAFASLLNY VGSDDLQGKRVGLFSYGSGLAASLYSCKIVGDVQHIIKELDITNKLAKRITETPKDYEAAIELRENAHLKKNFKPQGSIE HLQSGVYYLTNIDDKFRRSYDVKK

Truncated HMG-CoA Reductase

MVAVRRKALSILAEAPVLASDRLPYKNYDYDRVFGACCENVIGYMPLPVGVIGPLVIDGTSYHIPMATTEGCLVASAM RGCKAINAGGGATTVLTKDGMTRGPWRFPTLKRSGACKIWLDSEEGQNAIKKAFNSTSRFARLQHIQTCLAGDLLF MRFRTTTGDAMGMNMISKGVEYSLKQMVEEYGWEDMEWSVSGNYCTDKKPAAINWIEGRGKSWAEATIPGDW RKVLKSDVSALVELNIAKNLVGSAMAGSVGGFNAHAANLVTAVFLALGQDPAQNVESSNCITLMKEVDGDLRISVSM PSIEVGTIGGGTVLEPQGAMLDLLGVRGPHATAPGTNARQLARIVACAVLAGELSLCAALAAGHLVQSHMTHNR

Mevalonate Kinase

MSLPFLTSAPGKVIIFGEHSAVYNKPAVAASVSALRTYLLISESSAPDTIELDFPDISFNHKWSINDFNAITEDQVNSQK LAKAQQATDGLSQELVSLLDPLLAQLSESFHYHAAFCFLYMFVCLCPHAKNIKFSLKSTLPIGAGLGSSASISVSLALA MAYLGGLIGSNDLEKLSENDKHIVNQWAFIGEKCIHGTPSGIDNAVATYGNALLFEKDSHNGTINTNNFKFLDDFPAIP MILTYTRIPRSTKDLVARVRVLVTEKFPEVMKPILDAMGECALQGLEIMTKLSKCKGTDDEAVETNNELYEQLLELIRIN HGLLVSIGVSHPGLELIKNLSDDLRIGSTKLTGAGGGGCSLTLLRRDITQEQIDSFKKKLQDDFSYETFETDLGGTGCC LLSAKNLNKDLKIKSLVFQLFENKTTTKQQIDDLLLPGNTNLPWTS

Phosphomevalonate Kinase

MSELRAFSAPGKALLAGGYLVLDTKYEAFVVGLSARMHAVAHPYGSLQGSDKFEVRVKSKQFKDGEWLYHISPKSG FIPVSIGGSKNPFIEKVIANVFSYFKPNMDDYCNRNLFVIDIFSDDAYHSQEDSVTEHRGNRRLSFHSHRIEEVPKTGL GSSAGG[J1]LVTVLTTALASFFVSDLENNVDKYREVIHNLAQVAHCQAQGKIGSGFDVAAAAYGSIRYRRFPPALISNL PDIGSATYGSKLAHLVDEEDWNITIKSNHLPSGLTLWMGDIKNGSETVKLVQKVKNWYDSHMPESLKIYTELDHANS RFMDGLSKLDRLHETHDDYSDQIFESLERNDCTCQKYPEITEVRDAVATIRRSFRKITKESGADIEPPVQTSLLDDCQ TLKGVLTCLIPGAGGYDAIAVITKQDVDLRAQTANDKRFSKVQWLDVTQADWGVRKEKDPETYLDK

Figure 6A (continued)

Diphosphomevalonate Decarboxylase

MTVYTASVTAPVNIATLKYWGKRDTKLNLPTNSSISVTLSQDDLRTLTSAATAPEFERDTLWLNGEPHSIDNERTQNC LRDLRQLRKEMESKDASLPTLSQWKLHIVSENNFPTAAGLASSAAGFAALVSAIAKLYQLPQSTSEISRIARKGSGSA CRSLFGGYVAWEMGKAEDGHDSMAVQIADSSDWPQMKACVLWSDIKKDVSSTQGMQLTVATSELFKERIEHWP KRFEVMRKAIVEKDFATFAKETMMDSNSFHATCLDSFPPIFYMNDTSKRIISWCHTINQFYGETIVAYTFDAGPNAVL YYLAENESKLFAFIYKLFGSVPGWDKKFTTEQLEAFNHQFESSNFTARELDLELQKDVARVILTQVGSGPQETNESLI DAKTGLPKE

Isopentenyl-Diphosphate Delta-lsomerase

MTADNNSMPHGAVSSYAKLVQNQTPEDILEEFPEIIPLQQRPNTRSSETSNDESGETCFSGHDEEQIKLMNENCIVL DWDDNAIGAGTKKVCHLMENIEKGLLHRAFSVFIFNEQGELLLQQRATEKITFPDLWTNTCCSHPLCIDDELGLKGKL DDKIKGAITAAVRKLDHELGIPEDETKTRGKFHFLNRIHYMAPSNEPWGEHEIDYILFYKINAKENLTVNPNVNEVRDF KWVSPNDLKTMFADPSYKFTPWFKIICENYLFNVWVEQLDDLSEVENDRQIHRML

Geranyl-Diphosphate Synthase (ERG20ww)

MEAKIDELINNDPVWSSQNESLISKPYNHILLKPGKNFRLNLIVQINRVMNLPKDQLAIVSQIVELLHNSSLLIDDIEDNA PLRRGQTTSHLIWGVPSTINTANYMYFRAMQLVSQLTTKEPLYHWLITIFNEELINLHRGQGLDIYWRDFLPEIIPTQE MYLNMVMNKTGGLFRLTLRLMEALSPSSHHGHSLVPFINLLGIIYQIRDDYLNLKDFQMSSEKGFAEDITEGKLSFPIV HALNFTKTKGQTEQHNEILRILLLRTSDKDIKLKLIQILEFDTNSLAYTKNFINQLVNMIKNDNENKYLPDLASHSDTATN LHDELLYIIDHLSEL

Olivetol Synthase

MNHLRAEGPASVLAIGTANPENILLQDEFPDYYFRVTKSEHMTQLKEKFRKICDKSMIRKRNCFLNEEHLKQNPRLVE HEMQTLDARQDMLWEVPKLGKDACAKAIKEWGQPKSKITHLIFTSASTTDMPGADYHCAKLLGLSPSVKRVMMYQ LGCYGGGTVLRIAKDIAENNKGARVLAVCCDIMACLFRGPSESDLELLVGQAIFGDGAAAVIVGAEPDESVGERPIFE LVSTGQTILPNSEGTIGGHIREAGLIFDLHKDVPMLISNNIEKCLIEAFTPIGISDWNSIFWITHPGGKAILDKVEEKLHLK SDKFVDSRHVLSEHGNMSSSTVLFVMDELRKRSLEEGKSTTGDGFEWGVLFGFGPGLTVERVWRSVPIKY

Olivetolic Acid Cyclase

MAVKHLIVLKFKDEITEAQKEEFFKTYVNLVNIIPAMKDVYWGKDVTQKNKEEGYTHIVEVTFESVETIQDYIIHPAHVG FGDVYRSFWEKLLIFDYTPRK

CBGA Synthase

MGLSSVCTFSFQTNYHTLLNPHNNNPKTSLLCYRHPKTPIKYSYNNFPSKHCSTKSFHLQNKCSESLSIAKNSIRAAT TNQTEPPESDNHSVATKILNFGKACWKLQRPYTIIAFTSCACGLFGKELLHNTNLISWSLMFKAFFFLVAILCIASFTTTI NQIYDLHIDRINKPDLPLASGEISVNTAWIMSIIVALFGLIITIKMKGGPLYIFGYCFGIFGGIVYSVPPFRWKQNPSTAFL LNFLAHIITNFTFYYASRAALGLPFELRPSFTFLLAFMKSMGSALALIKDASDVEGDTKFGISTLASKYGSRNLTLFCSG IVLLSYVAAILAGIIWPQAFNSNVMLLSHAILAFWLILQTRDFALTNYDPEAGRRFYEFMWKLYYAEYLVYVFI

Figure 6A (continued)

Acetyl-CoA Carboxylase

MSEESLFESSPQKMEYEITNYSERHTELPGHFIGLNTVDKLEESPLRDFVKSHGGHTVISKILIANNGIAAVKEIRSVRK WAYETFGDDRTVQFVAMATPEDLEANAEYIRMADQYIEVPGGTNNNNYANVDLIVDIAERADVDAVWAGWGHASE NPLLPEKLSQSKRKVIFIGPPGNAMRSLGDKISSTIVAQSAKVPCIPWSGTGVDTVHVDEKTGLVSVDDDIYQKGCCT SPEDGLQKAKRIGFPVMIKASEGGGGKGIRQVEREEDFIALYHQAANEIPGSPIFIMKLAGRARHLEVQLLADQYGTNI SLFGRDCSVQRRHQKIIEEAPVTIAKAETFHEMEKAAVRLGKLVGYVSAGTVEYLYSHDDGKFYFLELNPRLQVEHP TTEMVSGVNLPAAQLQIAMGIPMHRISDIRTLYGMNPHSASEIDFEFKTQDATKKQRRPIPKGHCTACRITSEDPNDG FKPSGGTLHELNFRSSSNVWGYFSVGNNGNIHSFSDSQFGHIFAFGENRQASRKHMWALKELSIRGDFRTTVEYLI KLLETEDFEDNTITTGWLDDLITHKMTAEKPDPTLAVICGAATKAFLASEEARHKYIESLQKGQVLSKDLLQTMFPVDF IHEGKRYKFTVAKSGNDRYTLFINGSKCDIILRQLSDGGLLIAIGGKSHTIYWKEEVAATRLSVDSMTTLLEVENDPTQL RTPSPGKLVKFLVENGEHIIKGQPYAEIEVMKMQMPLVSQENGIVQLLKQPGSTIVAGDIMAIMTLDDPSKVKHALPFE GMLPDFGSPVIEGTKPAYKFKSLVSTLENILKGYDNQVIMNASLQQLIEVLRNPKLPYSEWKLHISALHSRLPAKLDEQ MEELVARSLRRGAVFPARQLSKLIDMAVKNPEYNPDKLLGAWEPLADIAHKYSNGLEAHEHSIFVHFLEEYYEVEKL FNGPNVREENIILKLRDENPKDLDKVALTVLSHSKVSAKNNLILAILKHYQPLCKLSSKVSAIFSTPLQHIVELESKATAK VALQAREILIQGALPSVKERTEQIEHILKSSVVKVAYGSSNPKRSEPDLNILKDLIDSNYWFDVLLQFLTHQDPWTAA AAQVYIRRAYRAYTIGDIRVHEGVTVPIVEWKFQLPSAAFSTFPTVKSKMGMNRAVSVSDLSYVANSQSSPLREGILM AVDHLDDVDEILSQSLEVIPRHQSSSNGPAPDRSGSSASLSNVANVCVASTEGFESEEEILVRLREILDLNKQELINAS IRRITFMFGFKDGSYPKYYTFNGPNYNENETIRHIEPALAFQLELGRLSNFNIKPIFTDNRNIHVYEAVSKTSPLDKRFF TRGIIRTGHIRDDISIQEYLTSEANRLMSDILDNLEVTDTSNSDLNHIFINFIAVFDISPEDVEAAFGGFLERFGKRLLRLR VSSAEIRIIIKDPQTGAPVPLRALINNVSGYVIKTEMYTEVKNAKGEVWFKSLGKPGSMHLRPIATPYPVKEWLQPKRY KAHLMGTTYVYDFPELFRQASSSQWKNFSADVKLTDDFFISNELIEDENGELTEVEREPGANAIGMVAFKITVKTPEY PRGRQFWVANDITFKIGSFGPQEDEFFNKVTEYARKRGIPRIYLAANSGARIGMAEEIVPLFQVAWNDAANPDKGF QYLYLTSEGMETLKKFDKENSVLTERTVINGEERFVIKTIIGSEDGLGVECLRGSGLIAGATSRAYHDIFTITLVTCRSV GIGAYLVRLGQRAIQVEGQPIILTGAPAINKMLGREVYTSNLQLGGTQIMYNNGVSHLTAVDDLAGVEKIVEWMSYVP AKRNMPVPILETKDTWDRPVDFTPTNDETYDVRWMIEGRETESGFEYGLFDKGSFFETLSGWAKGVWGRARLGGI PLGVIGVETRTVENLIPADPANPNSAETLIQEPGQVWHPNSAFKTAQAINDFNNGEQLPMMILANWRGFSGGQRDM FNEVLKYGSFIVDALVDYKQPIIIYIPPTGELRGGSWWVDPTINADQMEMYADVNARAGVLEPQGMVGIKFRREKLL DTMNRLDDKYRELRSQLSNKSLAPEVHQQISKQLADRERELLPIYGQISLQFADLHDRSSRMVAKGVISKELEWTEA RRFFFWRLRRRLNEEYLIKRLSHQVGEASRLEKIARIRSWYPASVDHEDDRQVATWIEENYKTLDDKLKGLKLESFA QDLAKKIRSDHDNAIDGLSEVIKMLSTDDKEKLLKTLK

CBDA Synthase

MKCSTFSFWFVCKIIFFFFSFNIQTSIANPRENFLKCFSQYIPNNATNLKLVYTQNNPLYMSVLNSTIHNLRFTSDTTPK PLVIVTPSHVSHIQGTILCSKKVGLQIRTRSGGHDSEGMSYISQVPFVIVDLRNMRSIKIDVHSQTAWVEAGATLGEVY YVWNEKNENLSLAAGYCPTVCAGGHFGGGGYGPLMRNYGLAADNIIDAHLVNVHGKVLDRKSMGEDLFWALRGG GAESFGIIVAWKIRLVAVPKSTMFSVKKIMEIHELVKLVNKWQNIAYKYDKDLLLMTHFITRNITDNQGKNKTAIHTYFS SVFLGGVDSLVDLMNKSFPELGIKKTDCRQLSWIDTIIFYSGWNYDTDNFNKEILLDRSAGQNGAFKIKLDYVKKPIP ESVFVQILEKLYEEDIGAGMYALYPYGGIMDEISESAIPFPHRAGILYELWYICSWEKQEDNEKHLNWIRNIYNFMTPY VSKNPRLAYLNYRDLDIGINDPKNPNNYTQARIWGEKYFGKNFDRLVKVKTLVDPNNFFRNEQSIPPLPRHRH

CBCA Synthase

MNCSTFSFWFVCKIIFFFLSFNIQISIANPQENFLKCFSEYIPNNPANPKFIYTQHDQLYMSVLNSTIQNLRFTSDTTPK PLVIVTPSNVSHIQASILCSKKVGLQIRTRSGGHDAEGLSYISQVPFAIVDLRNMHTVKVDIHSQTAWVEAGATLGEVY YWINEMNENFSFPGGYCPTVGVGGHFSGGGYGALMRNYGLAADNIIDAHLVNVDGKVLDRKSMGEDLFWAIRGGG GENFGIIAACKIKLVWPSKATIFSVKKNMEIHGLVKLFNKWQNIAYKYDKDLMLTTHFRTRNITDNHGKNKTTVHGYF SSIFLGGVDSLVDLMNKSFPELGIKKTDCKELSWIDTTIFYSGWNYNTANFKKEILLDRSAGKKTAFSIKLDYVKKLIPE TAMVKILEKLYEEEVGVGMYVLYPYGGIMDEISESAIPFPHRAGIMYELWYTATWEKQEDNEKHINWVRSVYNFTTP YVSQNPRLAYLNYRDLDLGKTNPESPNNYTQARIWGEKYFGKNFNRLVKVKTKADPNNFFRNEQSIPPLPPRHH

Figure 6A (continued)

Hexanoyl-CoA Synthetase

MGKNYKSLDSWASDFIALGITSEVAETLHGRLAEIVCNYGAATPQTWINIANHILSPDLPFSLHQMLFYGCYKDFGPA PPAWIPDPEKVKSTNLGALLEKRGKEFLGVKYKDPISSFSHFQEFSVRNPEVYWRTVLMDEMKISFSKDPECILRRD DINNPGGSEWLPGGYLNSAKNCLNVNSNKKLNDTMIVWRDEGNDDLPLNKLTLDQLRKRVWLVGYALEEMGLEKG CAIAIDMPMHVDAVVIYLAIVLAGYVWSIADSFSAPEISTRLRLSKAKAIFTQDHIIRGKKRIPLYSRVVEAKSPMAIVIP CSGSNIGAELRDGDISWDYFLERAKEFKNCEFTAREQPVDAYTNILFSSGTTGEPKAIPWTQATPLKAAADGWSHLD IRKGDVIVWPTNLGWMMGPWLVYASLLNGASIALYNGSPLVSGFAKFVQDAKVTMLGWPSIVRSWKSTNCVSGYD WSTIRCFSSSGEASNVDEYLWLMGRANYKPVIEMCGGTEIGGAFSAGSFLQAQSLSSFSSQCMGCTLYILDKNGYP MPKNKPGIGELALGPVMFGASKTLLNGNHHDVYFKGMPTLNGEVLRRHGDIFELTSNGYYHAHGRADDTMNIGGIKI SSIEIERVCNEVDDRVFETTAIGVPPLGGGPEQLVIFFVLKDSNDTTIDLNQLRLSFNLGLQKKLNPLFKVTRWPLSSL PRTATNKIMRRVLRQQFSHFE

Figure 6B

ATP Citrate Lyase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID1

MSAKAISEQTGKELLYKFICTTSAIQNRFKYARVTPDTDWARLLQDHPWLLSQNLWKPDQLIKRRGKLGLVGVNLTL DGVKSWLKPRLGQEATVGKATGFLKNFLIEPFVPHSQAEEFYVCIYATREGDYVLFHHEGGVDVGDVDAKAQKLLV GVDEKLNPEDIKKHLLVHAPEDKKEILASFISGLFNFYEDLYFTYLEINPLWTKDGVYVLDLAAKVDATADYICKVKWG DIEFPPPFGREAYPEEAYIADLDAKSGASLKLTLLNPKGRIWTMVAGGGASWYSDTICDLGGVNELANYGEYSGAP SEQQTYDYAKTILSLMTREKHPDGKILIIGGSIANFTNVAATFKGIVRAIRDYQGPLKEHEVTIFVRRGGPNYQEGLRV MGEVGKTTGIPIHVFGTETHMTAIVGMALGHRPIPNQPPTAAHTANFLLNASGSTSTPAPSRTASFSESRADEVAPAK KAKPAMPQDSVPSPRSLQGKSTTLFSRHTKAIVWGMQTRAVQGMLDFDYVCSRDEPSVAAMVYPFTGDHKQKFY WGHKEILIPVFKNMADAMRKHPEVDVLINFASLRSAYDSTMETMNYAQIRTIAIIAEGIPEALTRKLIKKADQKGVTIIGP ATVGGIKPGCFKIGNTGGMLDNILASKLYRPGSVAYVSRSGGMSNELNNIISRTTDGVYEGVAIGGDRYPGSTFMDH VLRYQDTPGVKMIWLGEIGGTEEYKICRGIKEGRLTKPIVCWCIGTCATMFSSEVQFGHAGACANQASETAVAKNQ ALKEAGVFVPRSFDELGEIIQSVYEDLVANGVIVPAQEVPPPTVPMDYSWARELGLIRKPASFMTSICDERGQELIYA GMPITEVFKEEMGIGGVLGLLWFQKRLPKYSCQFIEMCLMVTADHGPAVSGAHNTIICARAGKDLVSSLTSGLLTIGD RFGGALDAAAKMFSKAFDSGIIPMEFVNKMKKEGKLIMGIGHRVKSINNPDMRVQILKDYVRQHFPATPLLDYALEVE KITTSKKPNLILNVDGLIGVAFVDMLRNCGSFTREEADEYIDIGALNGIFVLGRSMGFIGHYLDQKRLKQGLYRHPWD DISYVLPEHMSMKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNA YYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQG DYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIE DYQKALELDPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENE TLKKKNLHKKDLIAYLEKEIANLRKKIEEGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHH HLESTSLYKKAGSGSARNAYLRKKIARLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASHEQ

Acetyl-CoA Acetyltransferase (atoB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID2

MKNCVIVSAVRTAIGSFNGSLASTSAIDLGATVIKAAIERAKIDSQHVDEVIMGNVLQAGLGQNPARQALLKSGLAETV CGFTVNKVCGSGLKSVALAAQAIQAGQAQSIVAGGMENMSLAPYLLDAKARSGYRLGDGQVYDVILRDGLMCATH GYHMGITAENVAKEYGITREMQDELALHSQRKAAAAIESGAFTAEIVPVNWTRKKTFVFSQDEFPKANSTAEALGAL RPAFDKAGTVTAGNASGINDGAAALVIMEESAALAAGLTPLARIKSYASGGVPPALMGMGPVPATQKALQLAGLQLA DIDLIEANEAFAAQFLAVGKNLGFDSEKVNVNGGAIALGHPIGASGARILVTLLHAMQARDKTLGLATLCIGGGQGIAM VIERLNKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGD YQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIE YYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALE LDPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSNEVTTLENDAAFIENENAYLEKEIAR LRKEKAALRNRLAHKKGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKA GSGSQKVAELKNRVAVKLNRNEQLKNKVEELKNRNAYLKNELATLENEVARLENDVAE

3-Hydroxybutyryl-CoA Dehydrogenase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID3

MKKVCVIGAGTMGSGIAQAFAAKGFEVVLRDIKDEFVDRGLDFINKNLSKLVKKGKIEEATKVEILTRISGTVDLNMAADCDL VIEAAVERMDIKKQIFADLDNICKPETILASNTSSLSITEVASATKRPDKVIGMHFFNPAPVMKLVEVIRGIATSQETFDAVKET SIAIGKDPVEVAEAPGFVVNRILIPMINEAVGILAEGIASVEDIDKAMKLGANHPMGPLELGDFIGLDICLAIMDVLYSETGDSK YRPHTLLKKYVRAGWLGRKSGKGFYDYSKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALEL DPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAW KNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQ GDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASENLYFQGENLYFQGDSSESCWNCGRKASETCSGCNT ARYCGSFCQHKDWEKHHHICGQTLQAQQGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSMAVSESQLK KMVSKYKYRDLTVRETVNVITLYKDLKPVLDSYVFNDGSSRELMNLTGTIPVPYRGNTYNIPICLWLLDTYPYNPPICFVKPT SSMTIKTGKHVDANGKIYLPYLHEWKHPQSDLLGLIQVMIVVFGDEPPVFSRP

Figure 6B (continued)

Enoyl-CoA Hydratase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID4

MELNNVILEKEGKVAWTINRPKALNALNSDTLKEMDYVIGEIENDSEVLAVILTGAGEKSFVAGADISEMKEMNTIEG RKFGILGNKVFRRLELLEKPVIAAVNGFALGGGCEIAMSCDIRIASSNARFGQPEVGLGITPGFGGTQRLSRLVGMGM AKQLIFTAQNIKADEALRIGLVNKWEPSELMNTAKEIANKIVSNAPVAVKLSKQAINRGMQCDIDTALAFESEAFGECF STEDQKDAMTAFIEKRKIEGFKNRKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPN NAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAW KNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAY YKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASGPLGSPLTASMLASAPPQEQKQMLGERLFP LIQAMHPTLAGKITGMLLEIDNSELLHMLESPESLRSKVDEAVAVLQAHQAKEAAQKAGSAGSAAGSGEFGSAEAAA KEAAAKAGSAGSAAGSGEFGSNTNMSVPTDGAVTTSQIPASEQETLVRPKPLLLKLLKSVGAQKDTYTMKEVLFYLG QYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLW

Trans-Enoyl-CoA Reductase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID5

MIVKPMVRNNICLNAHPQGCKKGVEDQIEYTKKRITAEVKAGAKAPKNVLVLGCSNGYGLASRITAAFGYGAATIGVS FEKAGSETKYGTPGWYNNLAFDEAAKREGLYSVTIDGDAFSDEIKAQVIEEAKKKGIKFDLIVYSLASPVRTDPDTGIM HKSVLKPFGKTFTGKTVDPFTGELKEISAEPANDEEAAATVKVMGGEDWERWIKQLSKEGLLEEGCITLAYSYIGPEA TQALYRKGTIGKAKEHLEATAHRLNKENPSIRAFVSVNKGLVTRASAVIPVIPLYLASLFKVMKEKGNHEGCIEQITRLY AERLYRKDGTIPVDEENRIRIDDWELEEDVQKAVSALMEKVTGENAESLTDLAGYRHDFLASNGFDVEGINYEAEVE RFDRIKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDY QKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEY YQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALEL DPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSNLLATLRSTAAVLENENHVLEKEKEKL RKEKEQLLNKLEAYKGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAG SGSKRIAYLRKKIAALKKDNANLEKDIANLENEIERLIKEIKTLENEVASHEQ

Beta-Ketothiolase (bktB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID6

MTREVWVSGVRTAIGTFGGSLKDVAPAELGALWREALARAQVSGDDVGHWFGNVIQTEPRDMYLGRVAAVNG GVTINAPALTVNRLCGSGLQAIVSAAQTILLGDTDVAIGGGAESMSRAPYLAPAARWGARMGDAGLVDMMLGALHD PFHRIHMGVTAENVAKEYDISRAQQDEAALESHRRASAAIKAGYFKDQIVPWSKGRKGDVTFDTDEHVRHDATIDD MTKLRPVFVKENGTVTAGNASGLNDAAAAVVMMERAEAERRGLKPLARLVSYGHAGVDPKAMGIGPVPATKIALER AGLQVSDLDVIEANEAFAAQACAVTKALGLDPAKVNPNGSGISLGHPIGATGALITVKALHELNRVQGRYALVTMCIG GGQGIAAIFERIKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAY YKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGD YQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIED YQKALELDPNNRSRSAGGGGSGGGGSGGGGASDVMWEYKWENTGDAELYGPFTSAQMQTWVSEGYFPDGVYC RKLDPPGGQFYNSKRIDFDLYTGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSESDSVEFNNAISYV NKIKTRFLDHPEIYRSFLEILHTYQKEQLHTKGRPFRGMSEEEVFTEVANLFRGQEDLLSEFGQFLPEAKR

Figure 6B (continued)

HMG-CoA Synthase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID7

MKLSTKLCWCGIKGRLRPQKQQQLHNTNLQMTELKKQKTAEQKTRPQNVGIKGIQIYIPTQCVNQSELEKFDGVSQ GKYTIGLGQTNMSFVNDREDIYSMSLTVLSKLIKSYNIDTNKIGRLEVGTETLIDKSKSVKSVLMQLFGENTDVEGIDTL NACYGGTNALFNSLNWIESNAWDGRDAIWCGDIAIYDKGAARPTGGAGTVAMWIGPDAPIVFDSVRASYMEHAYD FYKPDFTSEYPYVDGHFSLTCYVKALDQVYKSYSKKAISKGLVSDPAGSDALNVLKYFDYNVFHVPTCKLVTKSYGR LLYNDFRANPQLFPEVDAELATRDYDESLTDKNIEKTFVNVAKPFHKERVAQSLIVPTNTGNMYTASVYAAFASLLNY VGSDDLQGKRVGLFSYGSGLAASLYSCKIVGDVQHIIKELDITNKLAKRITETPKDYEAAIELRENAHLKKNFKPQGSIE HLQSGVYYLTNIDDKFRRSYDVKKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDP NNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEA WKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGN AYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASLGPLPPGWEVRSTVSGRIYFVDHNNRTT QFTDPRLHGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSGAMGPLPPGWEKRTDSNGRVYFVNH NTRITQWEDPRS

Truncated HMG-CoA Reductase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID8

MVAVRRKALSILAEAPVLASDRLPYKNYDYDRVFGACCENVIGYMPLPVGVIGPLVIDGTSYHIPMATTEGCLVASAM RGCKAINAGGGATTVLTKDGMTRGPWRFPTLKRSGACKIWLDSEEGQNAIKKAFNSTSRFARLQHIQTCLAGDLLF MRFRTTTGDAMGMNMISKGVEYSLKQMVEEYGWEDMEWSVSGNYCTDKKPAAINWIEGRGKSWAEATIPGDW RKVLKSDVSALVELNIAKNLVGSAMAGSVGGFNAHAANLVTAVFLALGQDPAQNVESSNCITLMKEVDGDLRISVSM PSIEVGTIGGGTVLEPQGAMLDLLGVRGPHATAPGTNARQLARIVACAVLAGELSLCAALAAGHLVQSHMTHNRKLS GGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYY QKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALE LDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRS RSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSEFFRRERNKMAAAKCRNRRRELTDTLQAETDQLE DEKSALQTEIANLLKEKEKLEFILAAHRPACKIPDDLGFPEEMSLEGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSA AGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVESLKQKIEELKQRKAQLKNDIANLEKEIAYAET

Mevalonate Kinase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID9

MSLPFLTSAPGKVIIFGEHSAVYNKPAVAASVSALRTYLLISESSAPDTIELDFPDISFNHKWSINDFNAITEDQVNSQK LAKAQQATDGLSQELVSLLDPLLAQLSESFHYHAAFCFLYMFVCLCPHAKNIKFSLKSTLPIGAGLGSSASISVSLALA MAYLGGLIGSNDLEKLSENDKHIVNQWAFIGEKCIHGTPSGIDNAVATYGNALLFEKDSHNGTINTNNFKFLDDFPAIP MILTYTRIPRSTKDLVARVRVLVTEKFPEVMKPILDAMGECALQGLEIMTKLSKCKGTDDEAVETNNELYEQLLELIRIN HGLLVSIGVSHPGLELIKNLSDDLRIGSTKLTGAGGGGCSLTLLRRDITQEQIDSFKKKLQDDFSYETFETDLGGTGCC LLSAKNLNKDLKIKSLVFQLFENKTTTKQQIDDLLLPGNTNLPWTSKLSGGGGSGGGGSGGGGSAEAWYNLGNAYY KQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQK AIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQ KALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASMEPAMEPET LEARINRATNPLNKELDWASINGFCEQLNEDFEGPPLATRLLAHKIQSPQEWEAIQALTVLETCMKSCGKRFHDEVG KFRFLNELIKWSPKYLGSRTSEKVKNKILELLYSWTVGLPEEVKIAEAYQMLKKQGIVKSGSAGSAAGSGEFGSAEA AAKEAAAKAGSAGSAAGSGEFGSGAMGSMAEAEGESLESWLNKATNPSNRQEDWEYIIGFCDQINKELEGPQIAV RLLAHKIQSPQEWEALQALTVLEACMKNCGRRFHNEVGKFRFLNELIKWSPKYLGDRVSEKVKTKVIELLYSWTMA LPEEAKIKDAYHMLKRQGIVQSDPPIPVDRTLIPSPPPRPKN

Figure 6B (continued)

Phosphomevalonate Kinase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID10

MSELRAFSAPGKALLAGGYLVLDTKYEAFVVGLSARMHAVAHPYGSLQGSDKFEVRVKSKQFKDGEWLYHISPKSG FIPVSIGGSKNPFIEKVIANVFSYFKPNMDDYCNRNLFVIDIFSDDAYHSQEDSVTEHRGNRRLSFHSHRIEEVPKTGL GSSAGG[J2]LVTVLTTALASFFVSDLENNVDKYREVIHNLAQVAHCQAQGKIGSGFDVAAAAYGSIRYRRFPPALISNL PDIGSATYGSKLAHLVDEEDWNITIKSNHLPSGLTLWMGDIKNGSETVKLVQKVKNWYDSHMPESLKIYTELDHANS RFMDGLSKLDRLHETHDDYSDQIFESLERNDCTCQKYPEITEVRDAVATIRRSFRKITKESGADIEPPVQTSLLDDCQ TLKGVLTCLIPGAGGYDAIAVITKQDVDLRAQTANDKRFSKVQWLDVTQADWGVRKEKDPETYLDKKLSGGGGSGG GGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDP NNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASA WYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGG SGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSQKVEELKNKIAELENRNAVKKNRVAHLKQEIAYLKDELAAH EFEGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSFENVTHEFIL ATLENENAKLRRLEAKLERELARLRNEVAWL

Diphosphomevalonate Decarboxylase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID11

MTVYTASVTAPVNIATLKYWGKRDTKLNLPTNSSISVTLSQDDLRTLTSAATAPEFERDTLWLNGEPHSIDNERTQNC LRDLRQLRKEMESKDASLPTLSQWKLHIVSENNFPTAAGLASSAAGFAALVSAIAKLYQLPQSTSEISRIARKGSGSA CRSLFGGYVAWEMGKAEDGHDSMAVQIADSSDWPQMKACVLWSDIKKDVSSTQGMQLTVATSELFKERIEHWP KRFEVMRKAIVEKDFATFAKETMMDSNSFHATCLDSFPPIFYMNDTSKRIISWCHTINQFYGETIVAYTFDAGPNAVL YYLAENESKLFAFIYKLFGSVPGWDKKFTTEQLEAFNHQFESSNFTARELDLELQKDVARVILTQVGSGPQETNESLI DAKTGLPKEKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYK QGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQ KAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQ KALELDPNNRSRSAGGGGSGGGGSGGGGASAMADLEQKVLEMEASTYDGVFIWKISDFPRKRQEAVAGRIPAIFS PAFYTSRYGYKMCLRIYLNGDGTGRGTHLSLFFWMKGPNDALLRWPFNQKVTLMLLDQNNREHVIDAFRPDVTSS SFQRPVNDMNIASGCPLFCPVSKMEAKNSYVRDDAIFIKAIVDLTGLGSAGSAAGSGEFGSAEAAAKEAAAKAGSAG SAAGSGEFGSASIKLQSSDGEIFEVDVEIAKQSVTIKTMLEDLGMDDEGDDDPVPLPNVNAAILKKVIQWCTHHKDDP PPPEDDENKEKRTDDIPVWDQEFLKVDQGTLFELILAANYLDIKGLLDVTCKTVANMIKGKTPEEIRKTFNIKNDFTEE EEAQVRKENQWC

Isopentenyl-Diphosphate Delta-lsomerase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID12

MTADNNSMPHGAVSSYAKLVQNQTPEDILEEFPEIIPLQQRPNTRSSETSNDESGETCFSGHDEEQIKLMNENCIVL DWDDNAIGAGTKKVCHLMENIEKGLLHRAFSVFIFNEQGELLLQQRATEKITFPDLWTNTCCSHPLCIDDELGLKGKL DDKIKGAITAAVRKLDHELGIPEDETKTRGKFHFLNRIHYMAPSNEPWGEHEIDYILFYKINAKENLTVNPNVNEVRDF KWVSPNDLKTMFADPSYKFTPWFKIICENYLFNVWVEQLDDLSEVENDRQIHRMLKLSGGGGSGGGGSGGGGSAE AWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGN AYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQ GDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGA SSYYHHHHHHLESTSLYKKAGSGSNTVKELKNYIQELEERNAELKNLKEHLKFAKAELEFELAAHKFEGSAGSAAGS GEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVAQLKNRVAYKLKENAKLE NIVARLENDNANLEKDIANLEKDIANLERDVAR

Figure 6B (continued)

Geranyl-Diphosphate Synthase (ERG20ww) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID13

MEAKIDELINNDPVWSSQNESLISKPYNHILLKPGKNFRLNLIVQINRVMNLPKDQLAIVSQIVELLHNSSLLIDDIEDNA PLRRGQTTSHLIWGVPSTINTANYMYFRAMQLVSQLTTKEPLYHWLITIFNEELINLHRGQGLDIYWRDFLPEIIPTQE MYLNMVMNKTGGLFRLTLRLMEALSPSSHHGHSLVPFINLLGIIYQIRDDYLNLKDFQMSSEKGFAEDITEGKLSFPIV HALNFTKTKGQTEQHNEILRILLLRTSDKDIKLKLIQILEFDTNSLAYTKNFINQLVNMIKNDNENKYLPDLASHSDTATN LHDELLYIIDHLSELKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLG NAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYK QGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQK AIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASLCTMKKGPSGYGFNLHSDKSKPGQFIRSVDPDSPAEAS GLRAQDRIVEVNGVCMEGKQHGDWSAIRAGGDETKLLVVDREGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSA AGSGEFGSSSGAIIYTVELKRYGGPLGITISGTEEPFDPIIISSLTKGGLAERTGAIHIGDRILAINSSSLKGKPLSEAIHLL QMAGETVTLKIKKQTDAQPASS

Olivetol Synthase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID14

MNHLRAEGPASVLAIGTANPENILLQDEFPDYYFRVTKSEHMTQLKEKFRKICDKSMIRKRNCFLNEEHLKQNPRLVE HEMQTLDARQDMLWEVPKLGKDACAKAIKEWGQPKSKITHLIFTSASTTDMPGADYHCAKLLGLSPSVKRVMMYQ LGCYGGGTVLRIAKDIAENNKGARVLAVCCDIMACLFRGPSESDLELLVGQAIFGDGAAAVIVGAEPDESVGERPIFE LVSTGQTILPNSEGTIGGHIREAGLIFDLHKDVPMLISNNIEKCLIEAFTPIGISDWNSIFWITHPGGKAILDKVEEKLHLK SDKFVDSRHVLSEHGNMSSSTVLFVMDELRKRSLEEGKSTTGDGFEWGVLFGFGPGLTVERVWRSVPIKYKLSGG GGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKA LELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDP NNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSA GGGGSGGGGSGGGGASGNNLETYEWYNKSISRDKAEKLLLDTGKEGAFMVRDSRTPGTYTVSVFTKAIISENPCIK HYHIKETNDSPKRYYVAEKYVFDSIPLLIQYHQYNGGGLVTRLRYPVCGGSAGSAAGSGEFGSAEAAAKEAAAKAG SAGSAAGSGEFGSGSHPWFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGK YFLWWKFNSLNELVDYHRSTSVSRNQQIFLRDIEQVPQQPT

Olivetolic Acid Cyclase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID15

MAVKHLIVLKFKDEITEAQKEEFFKTYVNLVNIIPAMKDVYWGKDVTQKNKEEGYTHIVEVTFESVETIQDYIIHPAHVG FGDVYRSFWEKLLIFDYTPRKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNA EAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKN LGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYK QGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASGQDRSEATLIKRFKGEGVRYKAKLIGIDEVSAA RGDKLCQDSMMKLKGWAGARSKGEHKQKIFLTISFGGIKIFDEKTGALQHHHAVHEISYIAKDITDHRAFGYVCGKE GNHRFVAIKTAQAAEPVILDLRDLFQLIYELKQREELEKKAGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSG EFGSGSHMGSQFWVTSQKTEASERCGLQGSYILRVEAEKLTLLTLGAQSQILEPLLFWPYTLLRRYGRDKVMFSFE AGRRCPSGPGTFTFQTSQGNDIFQAVEAAIQQQKAQGKVGQAQDILRLEHHHHHH

Figure 6B (continued)

CBGA Synthase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID16

MGLSSVCTFSFQTNYHTLLNPHNNNPKTSLLCYRHPKTPIKYSYNNFPSKHCSTKSFHLQNKCSESLSIAKNSIRAAT TNQTEPPESDNHSVATKILNFGKACWKLQRPYTilAFTSCACGLFGKELLHNTNLiSWSLMFKAFFFLVAiLCIASFTTTi NQlYDLHIDRINKPDLPLASGEISVNTAWIMSIIVALFGLilTIKMKGGPLYlFGYCFGlFGGIVYSVPPFRWKQNPSTAFL LNFLAHHTNFTFYYASRAALGLPFELRPSFTFLLAFMKSMGSALALiKDASDVEGDTKFGiSTLASKYGSRNLTLFCSG IVLLSYVAAILAGIIWPQAFNSNVMLLSHAILAFWLILQTRDFALTNYDPEAGRRFYEFMWKLYYAEYLVYVFIKLSGGG GSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKAL ELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPN NASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAG GGGSGGGGSGGGGASAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQVWVNAEDSEGKRGMIPVPYVEKYGS AGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSLIKHMRAEALFDFTGNSKLELNFKAGDVIFLLSRINKDW LEGTVRGATGIFPLSFVKILK

Acetyl-CoA Carboxylase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID17

MSEESLFESSPQKMEYEITNYSERHTELPGHFIGLNTVDKLEESPLRDFVKSHGGHTVISKILIANNGIAAVKEIRSVRK WAYETFGDDRTVQFVAMATPEDLEANAEYIRMADQYIEVPGGTNNNNYANVDLIVDIAERADVDAVWAGWGHASE NPLLPEKLSQSKRKVIFIGPPGNAMRSLGDKISSTIVAQSAKVPCIPWSGTGVDTVHVDEKTGLVSVDDDIYQKGCCT SPEDGLQKAKRIGFPVMIKASEGGGGKGIRQVEREEDFIALYHQAANEIPGSPIFIMKLAGRARHLEVQLLADQYGTNI SLFGRDCSVQRRHQKIIEEAPVTIAKAETFHEMEKAAVRLGKLVGYVSAGTVEYLYSHDDGKFYFLELNPRLQVEHP TTEMVSGVNLPAAQLQIAMGIPMHRISDIRTLYGMNPHSASEIDFEFKTQDATKKQRRPIPKGHCTACRITSEDPNDG FKPSGGTLHELNFRSSSNVWGYFSVGNNGNIHSFSDSQFGHIFAFGENRQASRKHMVVALKELSIRGDFRTTVEYLI KLLETEDFEDNTITTGWLDDLITHKMTAEKPDPTLAVICGAATKAFLASEEARHKYIESLQKGQVLSKDLLQTMFPVDF IHEGKRYKFTVAKSGNDRYTLFINGSKCDIILRQLSDGGLLIAIGGKSHTIYWKEEVAATRLSVDSMTTLLEVENDPTQL RTPSPGKLVKFLVENGEHIIKGQPYAEIEVMKMQMPLVSQENGIVQLLKQPGSTIVAGDIMAIMTLDDPSKVKHALPFE GMLPDFGSPVIEGTKPAYKFKSLVSTLENILKGYDNQVIMNASLQQLIEVLRNPKLPYSEWKLHISALHSRLPAKLDEQ MEELVARSLRRGAVFPARQLSKLIDMAVKNPEYNPDKLLGAWEPLADIAHKYSNGLEAHEHSIFVHFLEEYYEVEKL FNGPNVREENIILKLRDENPKDLDKVALTVLSHSKVSAKNNLILAILKHYQPLCKLSSKVSAIFSTPLQHIVELESKATAK VALQAREILIQGALPSVKERTEQIEHILKSSVVKVAYGSSNPKRSEPDLNILKDLIDSNYWFDVLLQFLTHQDPWTAA AAQVYIRRAYRAYTIGDIRVHEGVTVPIVEWKFQLPSAAFSTFPTVKSKMGMNRAVSVSDLSYVANSQSSPLREGILM AVDHLDDVDEILSQSLEVIPRHQSSSNGPAPDRSGSSASLSNVANVCVASTEGFESEEEILVRLREILDLNKQELINAS IRRITFMFGFKDGSYPKYYTFNGPNYNENETIRHIEPALAFQLELGRLSNFNIKPIFTDNRNIHVYEAVSKTSPLDKRFF TRGIIRTGHIRDDISIQEYLTSEANRLMSDILDNLEVTDTSNSDLNHIFINFIAVFDISPEDVEAAFGGFLERFGKRLLRLR VSSAEIRIIIKDPQTGAPVPLRALINNVSGYVIKTEMYTEVKNAKGEVWFKSLGKPGSMHLRPIATPYPVKEWLQPKRY KAHLMGTTYVYDFPELFRQASSSQWKNFSADVKLTDDFFISNELIEDENGELTEVEREPGANAIGMVAFKITVKTPEY PRGRQFWVANDITFKIGSFGPQEDEFFNKVTEYARKRGIPRIYLAANSGARIGMAEEIVPLFQVAWNDAANPDKGF QYLYLTSEGMETLKKFDKENSVLTERTVINGEERFVIKTIIGSEDGLGVECLRGSGLIAGATSRAYHDIFTITLVTCRSV GIGAYLVRLGQRAIQVEGQPIILTGAPAINKMLGREVYTSNLQLGGTQIMYNNGVSHLTAVDDLAGVEKIVEWMSYVP AKRNMPVPILETKDTWDRPVDFTPTNDETYDVRWMIEGRETESGFEYGLFDKGSFFETLSGWAKGVWGRARLGGI PLGVIGVETRTVENLIPADPANPNSAETLIQEPGQVWHPNSAFKTAQAINDFNNGEQLPMMILANWRGFSGGQRDM FNEVLKYGSFIVDALVDYKQPIIIYIPPTGELRGGSWWVDPTINADQMEMYADVNARAGVLEPQGMVGIKFRREKLL DTMNRLDDKYRELRSQLSNKSLAPEVHQQISKQLADRERELLPIYGQISLQFADLHDRSSRMVAKGVISKELEWTEA RRFFFWRLRRRLNEEYLIKRLSHQVGEASRLEKIARIRSWYPASVDHEDDRQVATWIEENYKTLDDKLKGLKLESFA QDLAKKIRSDHDNAIDGLSEVIKMLSTDDKEKLLKTLKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQK AIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKA LELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDP NNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASGSHMRLGAQSIQPTAN LDRTDDLVYLNVMELVRAVLELKNELAQLPPEGYWWKNVGLTLRKLIGSVDDLLPSLPSSSRTEIEGTQKLLNKDLA ELINKMRLAQQNAVTSLSEECKRQMLTASHTLAVDAKNLLDAVDQAKVLANLAHPPAEGSAGSAAGSGEFGSAEAA AKEAAAKAGSAGSAAGSGEFGSGAMATPGSENVLPREPLIATAVKFLQNSRVRQSPLATRRAFLKKKGLTDEEIDM AFQQSGTAADEPSSLW

Figure 6C

Cannabinoiderqic Metabolon Scaffold - (Mych

MGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSARNAYLRKKIA RLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSG EFSYYHHHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENETLKKKNLHKKDLIAYLEKEIANLRKKIEEGSA GSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQ KVAELKNRVAVKLNRNEQLKNKVEELKNRNAYLKNELATLENEVARLENDVAEGSAGSAAGSGEFAEAAAKEA AAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNEVTTLENDAAFIENENAYLEKEIARLRKEKAALR NRLAHKKGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSRPPTISNPPPLISSAK HPSVGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFNFLQSRPEPTAPPEESFRSGGSAGSAAGSGE FGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSKGTGLNPNAKVWQEIAPGNGSAGSAAGSGEF AEAAAKEAAAKAGSAGSAAGSGEFPDGGTTFEHLWSSLEPDSTYGSAGSAAGSGEFGSAEAAAKEAAAKEA AAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSKRIAYLRKKIAALKKDNANLEKDIANLE NEIERLIKEIKTLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLY KKAGSGSNLLATLRSTAAVLENENHVLEKEKEKLRKEKEQLLNKLEAYKGSAGSAAGSGEFGSAEAAAKEAAA KEAAAKEAAAKAGSAGSAAGSGEFGSPATSQHPPPPPGHRSQAPSHGSAGSAAGSGEFAEAAAKEAAAKAG SAGSAAGSGEFELNSLLILLEAAEYLERRDRGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSA AGSGEFGSRPPTISNPPPLISSAKHPSVGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFNFLQSRPEP TAPPEESFRSGGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSKGTGLNPNA KVWQEIAPGNGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFPDGGTTFEHLWSSLEPDSTYGSAGS AAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSKRIA YLRKKIAALKKDNANLEKDIANLENEIERLIKEIKTLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGS AAGSGEFSYYHHHHHHLESTSLYKKAGSGSNLLATLRSTAAVLENENHVLEKEKEKLRKEKEQLLNKLEAYKG SAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSALVDDAADYEPPPSNNEEALGSA GSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFRELFDDPSYVNVQNLDKARQGSAGSAAGSGEFGSAEAA AKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSKNTKSMNFDNPVYRKTTEEEGSAGSAAGSGEFAEAAAKE AAAKAGSAGSAAGSGEFRSLPSTWIENKLYGMSDPNWGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAA KAGSAGSAAGSGEFGSVVDNSPPPALPPKKRQSAPSGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGE FTQRSKPQPAVPPRPSADLILGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGST DEEREETEEEVYLLNSTTLGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFDGNVSGTQRLDSATVRT YSCGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKK AGSGSQKVAQLKNRVAYKLKENAKLENIVARLENDNANLEKDIANLEKDIANLERDVARGSAGSAAGSGEFAE AAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNTVKELKNYIQELEERNAELKNLKEHLK FAKAELEFELAAHKFEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSHDDSLP HPQQATDDSGHESDGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFGSPNAGSVEQTPKKPGLRRR GSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGS GSFENVTHEFILATLENENAKLRRLEAKLERELARLRNEVAWLGSAGSAAGSGEFAEAAAKEAAAKAGSAGSA AGSGEFSYYHHHHHHLESTSLYKKAGSGSQKVEELKNKIAELENRNAVKKNRVAHLKQEIAYLKDELAAHEFE GSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSVSSTKLVSFHDDSDEDLLHIGS AGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFAAATPISTFHDDSDEDLLHVGSAGSAAGSGEFGSAEAA AKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVESLKQKIEELKQRK AQLKNDIANLEKEIAYAETGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKK AGSEFFRRERNKMAAAKCRNRRRELTDTLQAETDQLEDEKSALQTEIANLLKEKEKLEFILAAHRPACKIPDDL GFPEEMSLEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSFQMPADTPPPAY LPPEDPMTGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFERESNEEPPPPYEDPYWGNGGSAGSA AGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVA ELKNRVAVKLNRNEQLKNKVEELKNRNAYLKNELATLENEVARLENDVAEGSAGSAAGSGEFAEAAAKEAAAK

Figure 6C (continued)

AGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNEVTTLENDAAFIENENAYLEKEIARLRKEKAALRNRL AHKKSYYHHHHHHLESTSLYKKAGSGSARNAYLRKKIARLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASH EQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNLVAQLENEV ASLENENETLKKKNLHKKDLIAYLEKEIANLRKKIEEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAG SAGSAAGSGEFGSEQKLISEEDLEQKLISEEDLEQKLISEEDLGSAGSAAGSGEFGSAGSAAGSGEFGSAGSA AGSGEF

Figure 6D

Malonyl-CoA Metabolon Scaffold - (FLAG)3

MGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSARNAYLRKKIA RLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSG EFSYYHHHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENETLKKKNLHKKDLIAYLEKEIANLRKKIEEGSA GSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSATRELDELMASLSDFKIQGGSAGS AAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFDLALSENWAQEFLAAGDAVDGSAGSAAGSGEFGSAEAAAK EAAAKEAAAKEAAAKAGSAGSAAGSGEFGSDYKDDDDKDYKDDDDKDYKDDDDKGSAGSAAGSGEFGSAG SAAGSGEFGSAGSAAGSGEF

<img file="IL283384A_D0161.tif" />

<img file="IL283384A_D0162.tif" />

CH, CH, OH O

<img file="IL283384A_D0163.tif" />

Cannabigerolic Acid

HoC 0 HO

OH

CH3

CH

H3C H

<img file="IL283384A_D0164.tif" />

<img file="IL283384A_D0165.tif" />

H

Ο^ΟΗ

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0166.tif" />

HO

Cannabigerolic Acid ch3

H2Q

<img file="IL283384A_D0167.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid ch3 ch3 oh o

H31

HO

Hexanoic Acid

<img file="IL283384A_D0168.tif" />

OH

CO

O I

CH3

CBDA Synthase ^(CBDAS)^

<img file="IL283384A_D0169.tif" />

ω

3,5,7-trioxodo- Olivetolic

Acid decanoyl-CoA

Geranyl Pyrophosphate ω

I ω

Cannabigerolic Acid

HexanoylC0A

HexanoylC0ASynthetase (HCS)

<img file="IL283384A_D0170.tif" />

Olivetol Synthase (OS)

Olivetolc Acid Cyclase K (OAC) ,

CBGA Synthase (CBGAS)

&amp;#1504;&amp;#1505;

JD14

&amp;#1504;&amp;#1505;

JD15

&amp;#1504;&amp;#1505;

JD16

<img file="IL283384A_D0171.tif" />

ID14 Scaffolded Ligands

ID15 Scaffolded Ligands

ID16 Scaffolded Ligands

Upper CB Pathway

FIG. 7 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0172.tif" />

E~

Δ hv

Δ ------> hv h3c

H

CH, C0 ch3

Cannabichromene

CH, OH

<img file="IL283384A_D0173.tif" />

H2Q h3c ho ch3

Cannabidiol

<img file="IL283384A_D0174.tif" />

Hexanoyl-CoA Synthetase

<img file="IL283384A_D0175.tif" />

CBCAS

Nucleus CH3 CH, OHO

A/xA/yUk!

H3C unAAz

Cannabigerolic Acid

Upper CB Pathway

<img file="IL283384A_D0176.tif" />

Upper CBergic Scaffold

CBDAS

<img file="IL283384A_D0177.tif" />

Cytosol ch3[

Δ >

hv

CH3 CH H&amp;#1470;C HO

<img file="IL283384A_D0178.tif" />

Cannabigerol

CH3

FIG. 7

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0179.tif" />

<img file="IL283384A_D0180.tif" />

CHo CHo OH O

<img file="IL283384A_D0181.tif" />

HO

Cannabigerolic Acid h3c

H

CH

OH

CH3

<img file="IL283384A_D0182.tif" />

H

O^OH θ^3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0183.tif" />

HO

Cannabigerolic Acid ch3

H2Q

<img file="IL283384A_D0184.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid ch3 ch3 oh o

H31

OH ch3

CBDA Synthase ^(CBDAS)^

<img file="IL283384A_D0185.tif" />

HO

Cannabigerolic Acid

<img file="IL283384A_D0186.tif" />

ω

IPP + DMAPP

Hexanoic Acid

HexanoylC0A

Geranyl Pyrophosphate

3,5,7-trioxodo- Olivetolic decanoyl-CoA Acid o O

I to

HexanoylC0ASynthetase &lt;(HCS)/

&amp;#1504;&amp;#1505;

IDX

IDX

Olivetol Synthase \ (OS) ,

&amp;#1504;&amp;#1505;

JD14

ID14

Olivetolic Acid Cyclase \(OAC)/

&amp;#1504;&amp;#1505;

JD15

ID15

CBGA Synthase (CBGAS)

&amp;#1499;&amp;#1505;

JD16

ID16

GPP Synthase (ERG20ww) F96W-N127W

&amp;#1504;&amp;#1505;

JD13

ID13 |2 Scaffolded Scaffolded Scaffolded Scaffolded Scaffolded

<img file="IL283384A_D0187.tif" />

Ligands

Ligands

Ligands

Ligands

Ligands

Truncated CBergic Scaffold

FIG. 8 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0188.tif" />

FIG. 8

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0189.tif" />

<img file="IL283384A_D0190.tif" />

ATP Citrate Acetyl-CoA

Lyase Carboxylase

<img file="IL283384A_D0191.tif" />

ATP Citrate Acetyl-CoA

Lyase Carboxylase

<img file="IL283384A_D0192.tif" />

Non-scaffolded Malonyl-CoA Pathway

<img file="IL283384A_D0193.tif" />

HO Ο

<img file="IL283384A_D0194.tif" />

ATP Citrate Lyase N (ACL) y

Acetyl-CoA AcetylT ransferase \(atoB)/

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

Enoyl-CoA Hydratase . (ECH) _

T rans-2enoyl-CoAReductase \(ECR)/

<img file="IL283384A_D0195.tif" />

Non-scaffolded Hexanoyl-CoA Pathway

<img file="IL283384A_D0196.tif" />

FIG. 9 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0197.tif" />

ATP Citrate Acetyl-CoA

Lyase (ACL)

Carboxylase (ACC)

<img file="IL283384A_D0198.tif" />

<img file="IL283384A_D0199.tif" />

Citrate Acetyl-CoA

<img file="IL283384A_D0200.tif" />

MalonylXi

Xi

Xi

Xi

Xi ω co

3-keto3-HydroxyT rans-Hex-2Hexanoyl-CoA Hexanoyl-CoA enoyl-CoA

HexanoylC0A

3,5,7trioxododecanoylC0A

Beta-ketothiolase (bktB)

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

Enoyl-CoA Hydratase . (ECH) '

T rans-2enoyl-CoAReductase \(ECR)/

Olivetol Synthase (OS)

<img file="IL283384A_D0201.tif" />

&amp;#1504;&amp;#1505;

JD14

<img file="IL283384A_D0202.tif" />

FIG. 9 (Continuous)

SUBSTITUTE SHEET (RULE 26)

ID14 Scaffolded Ligands

H

L

<img file="IL283384A_D0203.tif" />

<img file="IL283384A_D0204.tif" />

h3&amp;#1523;

CHo CHo OH O

<img file="IL283384A_D0205.tif" />

HO

Cannabigerolic Acid

OH

CH3

H3C H

CH

<img file="IL283384A_D0206.tif" />

H

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0207.tif" />

HO

Cannabigerolic Acid

CH3

H2Q

<img file="IL283384A_D0208.tif" />

<img file="IL283384A_D0209.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid ch3 ch3 oh o

ΗοΟ'^^Ο&amp;#972;&amp;#972; 3 HO^^

OH —CH3

CBDA Synthase . (CBDAS)

Cannabigerolic Acid

<img file="IL283384A_D0210.tif" />

IPP

MPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

R

S

CBGA Synthase (CBGAS)

Olivetol Acid Cyclase (OAC)

<img file="IL283384A_D0211.tif" />

o o

GPP Synthase (ERG20ww) F96W\N127W/

Isopentyl Diphosphate Isomerase \(IDI1)/

Diphosphomevalonate Decarboxylase \(MVD1)/

&amp;#1504;&amp;#1505;

JD15

&amp;#1504;&amp;#1505;

JD16

ID15 Scaffolded Ligands

ID16 Scaffolded Ligands

<img file="IL283384A_D0212.tif" />

Upper Cannabinoid Pathway

FIG. 9 (Continuous)

SUBSTITUTE SHEET (RULE 26)

X

<img file="IL283384A_D0213.tif" />

Cannabichromene

<img file="IL283384A_D0214.tif" />

<img file="IL283384A_D0215.tif" />

Mevalonate HMG-C0A

Δ >

hv cH3ch3 2H H3C IHO^

Cannabigerol

<img file="IL283384A_D0216.tif" />

Acetoacetyl- AcetylC0A C0A

<img file="IL283384A_D0217.tif" />

Mevalonate

phosphate

<img file="IL283384A_D0218.tif" />

<td> Phospho Mevalonate Kinase</td><td> Mevalonate Kinase (ERG12)</td><td> Truncated HMG-C0A Reductase</td><td> HMG-C0A Synthase (HMGS)</td><td> Acetyl-CoA AcetylT ransferase</td><td> ATP Citrate Lyase</td>

(ERG8L/\ ‘Z&amp;#1523;MtHMGRy\ / \(atoB)7\(ACL)

Non-scaffolded Mevalonate Pathway

FIG. 9

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0219.tif" />

<img file="IL283384A_D0220.tif" />

Acetoacetyl 3-Hydroxy- Trans-But Butanoyl-C0A Butanoyl-CoA enoyl-CoA C0A

<img file="IL283384A_D0221.tif" />

<img file="IL283384A_D0222.tif" />

—

Hexanoyl-CoA Pathway

®&amp;#1470; ©&amp;#1470;

T©

X©

FIG. 10 (Continuous)

SUBSTITUTE SHEET (RULE 26)

48/156 co co co co

3-KetoHexanoyl-CoA

3-HydroxyHexanoyl-CoA

T rans-Hex-2enoyl-CoA

HexanoylC0A

<img file="IL283384A_D0223.tif" />

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

Scaffolded

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

ID3 Scaffolded Ligands

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

<img file="IL283384A_D0224.tif" />

Citrate

ID4 Scaffolded Ligands

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

ID5 Scaffolded Ligands

Acetyl-CoA

<img file="IL283384A_D0225.tif" />

MalonylC0A

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0226.tif" />

o

ID17 Scaffolded Ligands

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

ID1 Scaffolded Ligands

3,5,7trioxododecanoylC0A

Olivetol Synthase (OS)

<img file="IL283384A_D0227.tif" />

&amp;#1504;&amp;#1505;

JD14

ID14 Scaffolded Ligands

I '— &amp;#1493; &lt;

Upper CB Pathway

MCA Pathway

L

FIG. 10 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0228.tif" />

<img file="IL283384A_D0229.tif" />

CH, CH, OH O

<img file="IL283384A_D0230.tif" />

Cannabigerolic Acid

HoC 0 HO

OH

CH3 h3c

H

CH

<img file="IL283384A_D0231.tif" />

H

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0232.tif" />

HO

Cannabigerolic Acid ch3

H2Q

<img file="IL283384A_D0233.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid

OH

CH3 CH3 OH Ο

<img file="IL283384A_D0234.tif" />

HO ch3

CBDA Synthase ^(CBDAS)^

<img file="IL283384A_D0235.tif" />

Cannabigerolic Acid

H31

<img file="IL283384A_D0236.tif" />

IPP

MPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

R

S

Olivetol Acid Cyclase . (OAC) ,

CBGA Synthase (CBGAS)

GPP Synthase (ERG20ww) F96W\N127W/

Isopentyl Diphosphate Isomerase \(IDI1)/

Diphosphomevalonate Decarboxylase \(MVD1)/ o

&amp;#1504;&amp;#1505;

ID15

<img file="IL283384A_D0237.tif" />

ID15 Scaffolded Ligands o

&amp;#1504;&amp;#1505;

ID16

<img file="IL283384A_D0238.tif" />

ID16 Scaffolded Ligands o

&amp;#1504;&amp;#1505;

ID13

<img file="IL283384A_D0239.tif" />

ID13 Scaffolded Ligands

&amp;#1504;&amp;#1505;

JD12

ID12 Scaffolded Ligands

&amp;#1504;&amp;#1505;

ID11

ID11 Scaffolded Ligands

FIG. 10 (Continuous)

SUBSTITUTE SHEET (RULE 26)

X

50/156 if a

Δ hv

Δ ------> hv

3U}=\CH3 H3C Q°

H0XV ch3

Cannabichromene

<img file="IL283384A_D0240.tif" />

CH, OH h2c

H3C HO ch Cannabidiol

R

<img file="IL283384A_D0241.tif" />

S

V

<img file="IL283384A_D0242.tif" />

Malonyl-CoA Pathway

Upper CB Pathway

<img file="IL283384A_D0243.tif" />

CBergic + HCA Pathway Scaffolds 13

<img file="IL283384A_D0244.tif" />

Mevalonate Pathway

Hexanoyl-CoA Pathway

Nucleus

<img file="IL283384A_D0245.tif" />

CH3 CHo OH Ο

Λν&amp;#1523;\Α/\ΑΑι h3c

Cannabigerolic Acid ch3

<img file="IL283384A_D0246.tif" />

Mevalonate , ± *

phosphate Mevalonate HMG-C0A

Cytosol hv o O to ch3 ch3

<img file="IL283384A_D0247.tif" />

Cannabigerol H&amp;#1470;C HO ch3 o o to

Acetoacetyl- AcetylC0A C0A

<img file="IL283384A_D0248.tif" />

Citrate

Phospho Mevalonate Truncated HMG-C0A Acetyl-CoA

Mevalonate

&amp;#1504;&amp;#1505;

JD1Q

ID10

Kinase

&amp;#1499;&amp;#1505;

ID9

ID9

HMG-C0A Synthase

&amp;#1499;&amp;#1505;

ID8

ID8

AcetylATP Citrate Lyase

&amp;#1504;&amp;#1505;

ID7

ID7

&amp;#1504;&amp;#1505;

ID2

ID2

&amp;#1504;&amp;#1505;

ID1

ID1

Scaffolded _ Scaffolded _ Scaffolded Scaffolded Scaffolded Scaffolded J

Ligands

Ligands

Ligands

Ligands

Ligands

Mevalonate Pathway

Ligands

<img file="IL283384A_D0249.tif" />

FIG. 10

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0250.tif" />

<img file="IL283384A_D0251.tif" />

Malonyl-CoA Pathway

<img file="IL283384A_D0252.tif" />

<img file="IL283384A_D0253.tif" />

<img file="IL283384A_D0254.tif" />

I

Hexanoyl-CoA Pathway

FIG. 11 (Continuous)

<img file="IL283384A_D0255.tif" />

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0256.tif" />

.ω ω co

3,5,7trioxododecanoylC0A

<img file="IL283384A_D0257.tif" />

MalonylCitrate Acetyl-CoA

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0258.tif" />

ο ο

&amp;#1502; &amp;#1504;&amp;#1505;

ID1

&amp;#1502; &amp;#1504;&amp;#1505;

JD1Z

HexanoylC0A

Olivetol Synthase (OS)

<img file="IL283384A_D0259.tif" />

&amp;#1504;&amp;#1505;

JD14

ID17 Scaffolded Ligands

ID1 Scaffolded Ligands (Λ (Λ (Λ

Τ rans-Hex Hydroxy3-ketoHexanoyl-CoA Hexanoyl-CoA enoyl-CoA

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

73 &amp;#1504;&amp;#1505;

ID3

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

ID6 Scaffolded

ID3 Scaffolded Ligands

ID4 Scaffolded Ligands

ID5 Scaffolded Ligands

ID14 Scaffolded Ligands

FIG. 11 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0260.tif" />

<img file="IL283384A_D0261.tif" />

h3&amp;#1523;

CH, CH, OH O

<img file="IL283384A_D0262.tif" />

HO

Cannabigerolic Acid

OH

CH3

H3C H

CH

<img file="IL283384A_D0263.tif" />

H

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0264.tif" />

HO

Cannabigerolic Acid ch3 ch3 oh o

ΗοΟ'^^Ο&amp;#972;&amp;#972; 3 HO^^

OH ch3

CH3

H2Q

<img file="IL283384A_D0265.tif" />

<img file="IL283384A_D0266.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid

CBDA Synthase ^(CBDAS)^

Cannabigerolic Acid

<img file="IL283384A_D0267.tif" />

IPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

R

S

Olivetol Acid Cyclase . (OAC) ,

&amp;#1504;&amp;#1505;

JD15

ID15 Scaffolded Ligands

MPP

CBGA Synthase (CBGAS)

&amp;#1504;&amp;#1505;

JD16

ID16 Scaffolded Ligands

GPP Synthase (ERG20ww) F96W\N127W/

Isopentyl Diphosphate Isomerase \(IDI1)/

Diphosphomevalonate Decarboxylase \(MVD1)/

&amp;#1504;&amp;#1505;

JD13

&amp;#1504;&amp;#1505;

JD12

&amp;#1504;&amp;#1505;

ID11

ID13 Scaffolded Ligands

ID12 Scaffolded Ligands

ID11 Scaffolded Ligands

Upper Cannabinoid Pathway

FIG. 11 (Continuous)

SUBSTITUTE SHEET (RULE 26) eg

Kx)

> &#1504;

Δ hv

Δ ------> hv 3>vCH3 H3C Q?

H0XV ch3

Cannabichromene

<img file="IL283384A_D0268.tif" />

Malonyl-CoA Pathway

Upper Cannabinoid Pathway

<img file="IL283384A_D0269.tif" />

CH, OH h2c h3c HO ch Cannabidiol

<img file="IL283384A_D0270.tif" />

CBergic + MCA Pathway Scaffolds 13

<img file="IL283384A_D0271.tif" />

Hexanoyl-CoA Pathway

Mevalonate Pathway

Nucleus

<img file="IL283384A_D0272.tif" />

<img file="IL283384A_D0273.tif" />

R

S CH3 CHoOHO

Λν,χΑζχΑΑι h3c

Cytosol

Cannabigerolic Acid ch3 hv

CH3 CH3

<img file="IL283384A_D0274.tif" />

Cannabigerol H&amp;#1470;C HO

CH3

<img file="IL283384A_D0275.tif" />

Mevalonate , ± *

phosphate Mevalonate HMG-C0A

Acetoacetyl- AcetylC0A C0A

<img file="IL283384A_D0276.tif" />

Citrate

Phospho Mevalonate Truncated HMG-C0A Acetyl-CoA

Mevalonate

&amp;#1504;&amp;#1505;

JD1Q

ID10

Kinase

&amp;#1499;&amp;#1505;

ID9

ID9

HMG-C0A Synthase

&amp;#1499;&amp;#1505;

ID8

ID8

AcetylATP Citrate Lyase

&amp;#1504;&amp;#1505;

ID7

ID7

&amp;#1504;&amp;#1505;

ID2

ID2

&amp;#1504;&amp;#1505;

ID1

ID1

Scaffolded _ Scaffolded _ Scaffolded Scaffolded Scaffolded Scaffolded J

Ligands

Ligands

Ligands

Ligands

Ligands

Ligands

Mevalonate Pathway

FIG. 11 (Continuous)

X

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0277.tif" />

&amp;#1504;&amp;#1506;

Acetyl-CoA

Acetyl-CoA

MalonylMalonylCitrate

Citrate

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0278.tif" />

ID1

ID17 Scaffolded Ligands

ID1 Scaffolded Ligands

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0279.tif" />

o o

&amp;#1502; &amp;#1504;&amp;#1505;

ID1

&amp;#1502; &amp;#1504;&amp;#1505;

ID1Z

ID1 Scaffolded Ligands

ID17 Scaffolded Ligands

Malonyl-CoA Pathway

HO Ο

<img file="IL283384A_D0280.tif" />

Citrate o O

&amp;#1504;&amp;#1506; o o

&amp;#1504;&amp;#1506; o

&amp;#1504;&amp;#1506;&amp;#1500; :ω o O

&amp;#1504;&amp;#1506;&amp;#1500;

Acetyl-CoA

Acetoacetyl

-C0A

o.&amp;#1488; 3-HydroxyButanoyl-CoA

Trans-But Butanoylenoyl-CoA C0A

ATP Citrate Lyase N (ACL) z

Acetyl-CoA AcetylT ransferase \(atoB)/

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

Enoyl-CoA Hydratase . (ECH) _

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

&amp;#1504;&amp;#1505;

ID4

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

&amp;#1502; &amp;#1504;&amp;#1505;

ID2

&amp;#1502; &amp;#1504;&amp;#1505;

ID1 £

g

Z

Z θ'

<td></td><td></td>

<td></td><td></td>

ID5 Scaffolded Ligands

ID4 Scaffolded Ligands

ID3 Scaffolded Ligands

ID2 Scaffolded Ligands

ID1 Scaffolded Ligands

Hexanoyl-CoA Pathway

<img file="IL283384A_D0281.tif" />

FIG. 11 (Continuous)

SUBSTITUTE SHEET (RULE 26)

<img file="IL283384A_D0282.tif" />

Xi

<img file="IL283384A_D0283.tif" />

MalonylCitrate Acetyl-CoA

ATP Citrate Lyase (ACL)

Acetyl-CoA Carboxylase (ACC)

<img file="IL283384A_D0284.tif" />

o o

&amp;#1502; &amp;#1504;&amp;#1505;

JD1Z

&amp;#1504;&amp;#1505;

ID1

U co

Xi b, o' e^

ID17 Scaffolded Ligands

ID1 Scaffolded Ligands

Xi

Xi

Xi

3,5,7trioxododecanoylC0A

HexanoylC0A

T rans-Hex Hydroxy3-ketoHexanoyl-CoA Hexanoyl-CoA enoyl-CoA

Olivetol Synthase (OS)

<img file="IL283384A_D0285.tif" />

&amp;#1504;&amp;#1505;

JD14

T rans-2enoyl-CoAReductase \(ECR)/

&amp;#1504;&amp;#1505;

ID5

ID5

ID14

<td> Scaffolded</td><td></td><td> Scaffolded</td>

Ligands

Ligands

Enoyl-CoA Hydratase . (ECH) '

&amp;#1502; &amp;#1504;&amp;#1505;

ID4

ID4 Scaffolded Ligands

3-hydroxybutyryl-CoA Dehydrogenase \(BHBD)/

&amp;#1502; &amp;#1504;&amp;#1505;

ID3

ID3 Scaffolded Ligands

FIG. 11 (Continuous)

SUBSTITUTE SHEET (RULE 26)

Beta-ketothiolase (bktB)

&amp;#1504;&amp;#1505;

ID6

ID6

Ligands

<img file="IL283384A_D0286.tif" />

<img file="IL283384A_D0287.tif" />

CHo CHo OH O

<img file="IL283384A_D0288.tif" />

Cannabigerolic Acid

HoC 0 HO

OH

CH3

CH

H3C H

<img file="IL283384A_D0289.tif" />

<img file="IL283384A_D0290.tif" />

H

O^OH CH3

Cannabichromenic Acid

CBCA Synthase (CBCAS)

<img file="IL283384A_D0291.tif" />

HO

Cannabigerolic Acid ch3

H2Q

<img file="IL283384A_D0292.tif" />

CH, OH O

OH

H3C HO ch3

Cannabidiolic Acid h3'

OH

CH3 CH3 OH O

<img file="IL283384A_D0293.tif" />

HO

CBDA Synthase ^(CBDAS)^

<img file="IL283384A_D0294.tif" />

Cannabigerolic Acid

CH3

<img file="IL283384A_D0295.tif" />

IPP

MPP

Olivetolic Acid

Geranyl

Pyrophosphate IPP + DMAPP

Olivetol Acid Cyclase . (OAC) ,

CBGA Synthase (CBGAS)

GPP Synthase (ERG20ww) F96W\N127W/

Isopentyl Diphosphate Isomerase \(IDI1)/

Diphosphomevalonate Decarboxylase \(MVD1)/

&amp;#1504;&amp;#1505;

JD15

&amp;#1504;&amp;#1505;

JD16

&amp;#1504;&amp;#1505;

JD13

&amp;#1504;&amp;#1505;

JD12

&amp;#1504;&amp;#1505;

ID11

ID15 Scaffolded Ligands

ID16 Scaffolded Ligands

ID13 Scaffolded Ligands

ID12 Scaffolded Ligands

ID11 _ Scaffolded j Ligands &amp;#1509;

Upper Cannabinoid Pathway

FIG. 11 (Continuous)

SUBSTITUTE SHEET (RULE 26)

Z.

<img file="IL283384A_D0296.tif" />

Cannabichromene

<img file="IL283384A_D0297.tif" />

Δ ------> hv

<img file="IL283384A_D0298.tif" />

CH, OH h2c

H3C HO ch Cannabidiol 13

Mitochondria or Plastid

Cytosol

Cannabigerolic Acid o Ui ch3 ch3 ch, OH ο

Δ hv

<img file="IL283384A_D0299.tif" />

Mevalonate. , ± , _ &amp;#1524;

phosphate Mevalonate HMG-C0A

Cytosol ch3 ch3

<img file="IL283384A_D0300.tif" />

Cannabigerol H&amp;#1470;C HO ch3

Acetoacetyl- AcetylC0A C0A

<img file="IL283384A_D0301.tif" />

Citrate

Phospho- Mevalonate Truncated HMG-C0A Acetyl-CoA

Mevalonate

I \ID1Q

Kinase

&amp;#1504;&amp;#1505;

ID9

ID9

HMG-C0A Synthase

&amp;#1504;&amp;#1505;

ID8

ID8

AcetylATP Citrate Lyase

&amp;#1504;&amp;#1505;

ID7

ID7

&amp;#1502; &amp;#1504;&amp;#1505;

ID2

&amp;#1504;&amp;#1505;

ID1

ID1

ID10 _ ID9 _ ID8 _ ID7 ID2 . ID1 (0¾ Scaffolded _ Scaffolded _ Scaffolded Scaffolded Scaffolded Scaffolded ] (nw&amp;#1470;| &amp;#2404; inondc Π &amp;#1493; inondo Π 1 inonric Π &amp;#1493; inondo Π Ligands Ligands @T Ligands

Ligands

Ligands

Ligands

Ligands

Mevalonate Pathway

<img file="IL283384A_D0302.tif" />

FIG. 11

SUBSTITUTE SHEET (RULE 26)

Figure 12A

ATP Citrate Lyase

ATGTCTGCTAAAGCTATTTCTGAACAAACTGGTAAAGAATTGTTGTATAAATTTATTTGTACTACTTCTGCTATTC AAAATAGATTTAAATATGCTAGAGTTACTCCAGATACTGATTGGGCTAGATTGTTGCAAGATCATCCATGGTTGT TGTCTCAAAATTTGGTTGTTAAACCAGATCAATTGATTAAAAGAAGAGGTAAATTGGGTTTGGTTGGTGTTAATTT GACTTTGGATGGTGTTAAATCTTGGTTGAAACCAAGATTGGGTCAAGAAGCTACTGTTGGTAAAGCTACTGGTT TTTTGAAAAATTTTTTGATTGAACCATTTGTTCCACATTCTCAAGCTGAAGAATTTTATGTTTGTATTTATGCTACT AGAGAAGGTGATTATGTTTTGTTTCATCATGAAGGTGGTGTTGATGTTGGTGATGTTGATGCTAAAGCTCAAAAA TTGTTGGTTGGTGTTGATGAAAAATTGAATCCAGAAGATATTAAAAAACATTTGTTGGTTCATGCTCCAGAAGAT AAAAAAGAAATTTTGGCTTCTTTTATTTCTGGTTTGTTTAATTTTTATGAAGATTTGTATTTTACTTATTTGGAAATT AATCCATTGGTTGTTACTAAAGATGGTGTTTATGTTTTGGATTTGGCTGCTAAAGTTGATGCTACTGCTGATTATA TTTGTAAAGTTAAATGGGGTGATATTGAATTTCCACCACCATTTGGTAGAGAAGCTTATCCAGAAGAAGCTTATA TTGCTGATTTGGATGCTAAATCTGGTGCTTCTTTGAAATTGACTTTGTTGAATCCAAAAGGTAGAATTTGGACTAT GGTTGCTGGTGGTGGTGCTTCTGTTGTTTATTCTGATACTATTTGTGATTTGGGTGGTGTTAATGAATTGGCTAA TTATGGTGAATATTCTGGTGCTCCATCTGAACAACAAACTTATGATTATGCTAAAACTATTTTGTCTTTGATGACT AGAGAAAAACATCCAGATGGTAAAATTTTGATTATTGGTGGTTCTATTGCTAATTTTACTAATGTTGCTGCTACTT TTAAAGGTATTGTTAGAGCTATTAGAGATTATCAAGGTCCATTGAAAGAACATGAAGTTACTATTTTTGTTAGAAG AGGTGGTCCAAATTATCAAGAAGGTTTGAGAGTTATGGGTGAAGTTGGTAAAACTACTGGTATTCCAATTCATGT TTTTGGTACTGAAACTCATATGACTGCTATTGTTGGTATGGCTTTGGGTCATAGACCAATTCCAAATCAACCACC AACTGCTGCTCATACTGCTAATTTTTTGTTGAATGCTTCTGGTTCTACTTCTACTCCAGCTCCATCTAGAACTGCT TCTTTTTCTGAATCTAGAGCTGATGAAGTTGCTCCAGCTAAAAAAGCTAAACCAGCTATGCCACAAGATTCTGTT CCATCTCCAAGATCTTTGCAAGGTAAATCTACTACTTTGTTTTCTAGACATACTAAAGCTATTGTTTGGGGTATGC AAACTAGAGCTGTTCAAGGTATGTTGGATTTTGATTATGTTTGTTCTAGAGATGAACCATCTGTTGCTGCTATGG TTTATCCATTTACTGGTGATCATAAACAAAAATTTTATTGGGGTCATAAAGAAATTTTGATTCCAGTTTTTAAAAAT ATGGCTGATGCTATGAGAAAACATCCAGAAGTTGATGTTTTGATTAATTTTGCTTCTTTGAGATCTGCTTATGATT CTACTATGGAAACTATGAATTATGCTCAAATTAGAACTATTGCTATTATTGCTGAAGGTATTCCAGAAGCTTTGAC TAGAAAATTGATTAAAAAAGCTGATCAAAAAGGTGTTACTATTATTGGTCCAGCTACTGTTGGTGGTATTAAACC AGGTTGTTTTAAAATTGGTAATACTGGTGGTATGTTGGATAATATTTTGGCTTCTAAATTGTATAGACCAGGTTCT GTTGCTTATGTTTCTAGATCTGGTGGTATGTCTAATGAATTGAATAATATTATTTCTAGAACTACTGATGGTGTTT ATGAAGGTGTTGCTATTGGTGGTGATAGATATCCAGGTTCTACTTTTATGGATCATGTTTTGAGATATCAAGATA CTCCAGGTGTTAAAATGATTGTTGTTTTGGGTGAAATTGGTGGTACTGAAGAATATAAAATTTGTAGAGGTATTA AAGAAGGTAGATTGACTAAACCAATTGTTTGTTGGTGTATTGGTACTTGTGCTACTATGTTTTCTTCTGAAGTTCA ATTTGGTCATGCTGGTGCTTGTGCTAATCAAGCTTCTGAAACTGCTGTTGCTAAAAATCAAGCTTTGAAAGAAGC TGGTGTTTTTGTTCCAAGATCTTTTGATGAATTGGGTGAAATTATTCAATCTGTTTATGAAGATTTGGTTGCTAAT GGTGTTATTGTTCCAGCTCAAGAAGTTCCACCACCAACTGTTCCAATGGATTATTCTTGGGCTAGAGAATTGGG TTTGATTAGAAAACCAGCTTCTTTTATGACTTCTATTTGTGATGAAAGAGGTCAAGAATTGATTTATGCTGGTATG CCAATTACTGAAGTTTTTAAAGAAGAAATGGGTATTGGTGGTGTTTTGGGTTTGTTGTGGTTTCAAAAAAGATTG CCAAAATATTCTTGTCAATTTATTGAAATGTGTTTGATGGTTACTGCTGATCATGGTCCAGCTGTTTCTGGTGCT CATAATACTATTATTTGTGCTAGAGCTGGTAAAGATTTGGTTTCTTCTTTGACTTCTGGTTTGTTGACTATTGGTG ATAGATTTGGTGGTGCTTTGGATGCTGCTGCTAAAATGTTTTCTAAAGCTTTTGATTCTGGTATTATTCCAATGGA ATTTGTTAATAAAATGAAAAAAGAAGGTAAATTGATTATGGGTATTGGTCATAGAGTTAAATCTATTAATAATCCA GATATGAGAGTTCAAATTTTGAAAGATTATGTTAGACAACATTTTCCAGCTACTCCATTGTTGGATTATGCTTTGG AAGTTGAAAAAATTACTACTTCTAAAAAACCAAATTTGATTTTGAATGTTGATGGTTTGATTGGTGTTGCTTTTGTT GATATGTTGAGAAATTGTGGTTCTTTTACTAGAGAAGAAGCTGATGAATATATTGATATTGGTGCTTTGAATGGT ATTTTTGTTTTGGGTAGATCTATGGGTTTTATTGGTCATTATTTGGATCAAAAAAGATTGAAACAAGGTTTGTATA GACATCCATGGGATGATATTTCTTATGTTTTGCCAGAACATATGTCTATG

Figure 12A (continued)

Acetyl-CoA Acetyltransferase (atoB)

ATGAAAAATTGTGTTATTGTTTCTGCTGTTAGAACTGCTATTGGTTCTTTTAATGGTTCTTTGGCTTCTACTTCTG CTATTGATTTGGGTGCTACTGTTATTAAAGCTGCTATTGAAAGAGCTAAAATTGATTCTCAACATGTTGATGAAGT TATTATGGGTAATGTTTTGCAAGCTGGTTTGGGTCAAAATCCAGCTAGACAAGCTTTGTTGAAATCTGGTTTGGC TGAAACTGTTTGTGGTTTTACTGTTAATAAAGTTTGTGGTTCTGGTTTGAAATCTGTTGCTTTGGCTGCTCAAGCT ATTCAAGCTGGTCAAGCTCAATCTATTGTTGCTGGTGGTATGGAAAATATGTCTTTGGCTCCATATTTGTTGGAT GCTAAAGCTAGATCTGGTTATAGATTGGGTGATGGTCAAGTTTATGATGTTATTTTGAGAGATGGTTTGATGTGT GCTACTCATGGTTATCATATGGGTATTACTGCTGAAAATGTTGCTAAAGAATATGGTATTACTAGAGAAATGCAA GATGAATTGGCTTTGCATTCTCAAAGAAAAGCTGCTGCTGCTATTGAATCTGGTGCTTTTACTGCTGAAATTGTT CCAGTTAATGTTGTTACTAGAAAAAAAACTTTTGTTTTTTCTCAAGATGAATTTCCAAAAGCTAATTCTACTGCTG AAGCTTTGGGTGCTTTGAGACCAGCTTTTGATAAAGCTGGTACTGTTACTGCTGGTAATGCTTCTGGTATTAATG ATGGTGCTGCTGCTTTGGTTATTATGGAAGAATCTGCTGCTTTGGCTGCTGGTTTGACTCCATTGGCTAGAATTA AATCTTATGCTTCTGGTGGTGTTCCACCAGCTTTGATGGGTATGGGTCCAGTTCCAGCTACTCAAAAAGCTTTG CAATTGGCTGGTTTGCAATTGGCTGATATTGATTTGATTGAAGCTAATGAAGCTTTTGCTGCTCAATTTTTGGCT GTTGGTAAAAATTTGGGTTTTGATTCTGAAAAAGTTAATGTTAATGGTGGTGCTATTGCTTTGGGTCATCCAATT GGTGCTTCTGGTGCTAGAATTTTGGTTACTTTGTTGCATGCTATGCAAGCTAGAGATAAAACTTTGGGTTTGGCT ACTTTGTGTATTGGTGGTGGTCAAGGTATTGCTATGGTTATTGAAAGATTGAAT

3-Hvdroxvbutvryl-CoA Dehydrogenase

ATGAAAAAAGTTTGTGTTATTGGTGCTGGTACTATGGGTTCTGGTATTGCTCAAGCTTTTGCTGCTAAAGGTTTT GAAGTTGTTTTGAGAGATATTAAAGATGAATTTGTTGATAGAGGTTTGGATTTTATTAATAAAAATTTGTCTAAATT GGTTAAAAAAGGTAAAATTGAAGAAGCTACTAAAGTTGAAATTTTGACTAGAATTTCTGGTACTGTTGATTTGAAT ATGGCTGCTGATTGTGATTTGGTTATTGAAGCTGCTGTTGAAAGAATGGATATTAAAAAACAAATTTTTGCTGATT TGGATAATATTTGTAAACCAGAAACTATTTTGGCTTCTAATACTTCTTCTTTGTCTATTACTGAAGTTGCTTCTGCT ACTAAAAGACCAGATAAAGTTATTGGTATGCATTTTTTTAATCCAGCTCCAGTTATGAAATTGGTTGAAGTTATTA GAGGTATTGCTACTTCTCAAGAAACTTTTGATGCTGTTAAAGAAACTTCTATTGCTATTGGTAAAGATCCAGTTG AAGTTGCTGAAGCTCCAGGTTTTGTTGTTAATAGAATTTTGATTCCAATGATTAATGAAGCTGTTGGTATTTTGGC TGAAGGTATTGCTTCTGTTGAAGATATTGATAAAGCTATGAAATTGGGTGCTAATCATCCAATGGGTCCATTGGA ATTGGGTGATTTTATTGGTTTGGATATTTGTTTGGCTATTATGGATGTTTTGTATTCTGAAACTGGTGATTCTAAA TATAGACCACATACTTTGTTGAAAAAATATGTTAGAGCTGGTTGGTTGGGTAGAAAATCTGGTAAAGGTTTTTAT GATTATTCTAAA

Enoyl-CoA Hydratase

ATGGAATTGAATAATGTTATTTTGGAAAAAGAAGGTAAAGTTGCTGTTGTTACTATTAATAGACCAAAAGCTTTGA ATGCTTTGAATTCTGATACTTTGAAAGAAATGGATTATGTTATTGGTGAAATTGAAAATGATTCTGAAGTTTTGGC TGTTATTTTGACTGGTGCTGGTGAAAAATCTTTTGTTGCTGGTGCTGATATTTCTGAAATGAAAGAAATGAATACT ATTGAAGGTAGAAAATTTGGTATTTTGGGTAATAAAGTTTTTAGAAGATTGGAATTGTTGGAAAAACCAGTTATTG CTGCTGTTAATGGTTTTGCTTTGGGTGGTGGTTGTGAAATTGCTATGTCTTGTGATATTAGAATTGCTTCTTCTAA TGCTAGATTTGGTCAACCAGAAGTTGGTTTGGGTATTACTCCAGGTTTTGGTGGTACTCAAAGATTGTCTAGATT GGTTGGTATGGGTATGGCTAAACAATTGATTTTTACTGCTCAAAATATTAAAGCTGATGAAGCTTTGAGAATTGG TTTGGTTAATAAAGTTGTTGAACCATCTGAATTGATGAATACTGCTAAAGAAATTGCTAATAAAATTGTTTCTAAT GCTCCAGTTGCTGTTAAATTGTCTAAACAAGCTATTAATAGAGGTATGCAATGTGATATTGATACTGCTTTGGCT TTTGAATCTGAAGCTTTTGGTGAATGTTTTTCTACTGAAGATCAAAAAGATGCTATGACTGCTTTTATTGAAAAAA GAAAAATTGAAGGTTTTAAAAATAGA

Figure 12A (continued)

Trans-Enoyl-CoA Reductase

ATGATTGTTAAACCAATGGTTAGAAATAATATTTGTTTGAATGCTCATCCACAAGGTTGTAAAAAAGGTGTTGAA GATCAAATTGAATATACTAAAAAAAGAATTACTGCTGAAGTTAAAGCTGGTGCTAAAGCTCCAAAAAATGTTTTG GTTTTGGGTTGTTCTAATGGTTATGGTTTGGCTTCTAGAATTACTGCTGCTTTTGGTTATGGTGCTGCTACTATT GGTGTTTCTTTTGAAAAAGCTGGTTCTGAAACTAAATATGGTACTCCAGGTTGGTATAATAATTTGGCTTTTGAT GAAGCTGCTAAAAGAGAAGGTTTGTATTCTGTTACTATTGATGGTGATGCTTTTTCTGATGAAATTAAAGCTCAA GTTATTGAAGAAGCTAAAAAAAAAGGTATTAAATTTGATTTGATTGTTTATTCTTTGGCTTCTCCAGTTAGAACTG ATCCAGATACTGGTATTATGCATAAATCTGTTTTGAAACCATTTGGTAAAACTTTTACTGGTAAAACTGTTGATCC ATTTACTGGTGAATTGAAAGAAATTTCTGCTGAACCAGCTAATGATGAAGAAGCTGCTGCTACTGTTAAAGTTAT GGGTGGTGAAGATTGGGAAAGATGGATTAAACAATTGTCTAAAGAAGGTTTGTTGGAAGAAGGTTGTATTACTT TGGCTTATTCTTATATTGGTCCAGAAGCTACTCAAGCTTTGTATAGAAAAGGTACTATTGGTAAAGCTAAAGAAC ATTTGGAAGCTACTGCTCATAGATTGAATAAAGAAAATCCATCTATTAGAGCTTTTGTTTCTGTTAATAAAGGTTT GGTTACTAGAGCTTCTGCTGTTATTCCAGTTATTCCATTGTATTTGGCTTCTTTGTTTAAAGTTATGAAAGAAAAA GGTAATCATGAAGGTTGTATTGAACAAATTACTAGATTGTATGCTGAAAGATTGTATAGAAAAGATGGTACTATT CCAGTTGATGAAGAAAATAGAATTAGAATTGATGATTGGGAATTGGAAGAAGATGTTCAAAAAGCTGTTTCTGCT TTGATGGAAAAAGTTACTGGTGAAAATGCTGAATCTTTGACTGATTTGGCTGGTTATAGACATGATTTTTTGGCT TCTAATGGTTTTGATGTTGAAGGTATTAATTATGAAGCTGAAGTTGAAAGATTTGATAGAATT

Beta-Ketothiolase (bktB)

ATGACTAGAGAAGTTGTTGTTGTTTCTGGTGTTAGAACTGCTATTGGTACTTTTGGTGGTTCTTTGAAAGATGTT GCTCCAGCTGAATTGGGTGCTTTGGTTGTTAGAGAAGCTTTGGCTAGAGCTCAAGTTTCTGGTGATGATGTTGG TCATGTTGTTTTTGGTAATGTTATTCAAACTGAACCAAGAGATATGTATTTGGGTAGAGTTGCTGCTGTTAATGG TGGTGTTACTATTAATGCTCCAGCTTTGACTGTTAATAGATTGTGTGGTTCTGGTTTGCAAGCTATTGTTTCTGCT GCTCAAACTATTTTGTTGGGTGATACTGATGTTGCTATTGGTGGTGGTGCTGAATCTATGTCTAGAGCTCCATAT TTGGCTCCAGCTGCTAGATGGGGTGCTAGAATGGGTGATGCTGGTTTGGTTGATATGATGTTGGGTGCTTTGC ATGATCCATTTCATAGAATTCATATGGGTGTTACTGCTGAAAATGTTGCTAAAGAATATGATATTTCTAGAGCTCA ACAAGATGAAGCTGCTTTGGAATCTCATAGAAGAGCTTCTGCTGCTATTAAAGCTGGTTATTTTAAAGATCAAAT TGTTCCAGTTGTTTCTAAAGGTAGAAAAGGTGATGTTACTTTTGATACTGATGAACATGTTAGACATGATGCTAC TATTGATGATATGACTAAATTGAGACCAGTTTTTGTTAAAGAAAATGGTACTGTTACTGCTGGTAATGCTTCTGGT TTGAATGATGCTGCTGCTGCTGTTGTTATGATGGAAAGAGCTGAAGCTGAAAGAAGAGGTTTGAAACCATTGGC TAGATTGGTTTCTTATGGTCATGCTGGTGTTGATCCAAAAGCTATGGGTATTGGTCCAGTTCCAGCTACTAAAAT TGCTTTGGAAAGAGCTGGTTTGCAAGTTTCTGATTTGGATGTTATTGAAGCTAATGAAGCTTTTGCTGCTCAAGC TTGTGCTGTTACTAAAGCTTTGGGTTTGGATCCAGCTAAAGTTAATCCAAATGGTTCTGGTATTTCTTTGGGTCA TCCAATTGGTGCTACTGGTGCTTTGATTACTGTTAAAGCTTTGCATGAATTGAATAGAGTTCAAGGTAGATATGC TTTGGTTACTATGTGTATTGGTGGTGGTCAAGGTATTGCTGCTATTTTTGAAAGAATT

Figure 12A (continued)

HMG CoA Synthase

ATGAAACTCTCAACTAAACTTTGTTGGTGTGGTATTAAAGGAAGACTTAGGCCGCAAAAGCAACAACAATTACAC AATACAAACTTGCAAATGACTGAACTAAAAAAACAAAAGACCGCTGAACAAAAAACCAGACCTCAAAATGTCGGT ATTAAAGGTATCCAAATTTACATCCCAACTCAATGTGTCAACCAATCTGAGCTAGAGAAATTTGATGGCGTTTCT CAAGGTAAATACACAATTGGTCTGGGCCAAACCAACATGTCTTTTGTCAATGACAGAGAAGATATCTACTCGAT GTCCCTAACTGTTTTGTCTAAGTTGATCAAGAGTTACAACATCGACACCAACAAAATTGGTAGATTAGAAGTCGG TACTGAAACTCTGATTGACAAGTCCAAGTCTGTCAAGTCTGTCTTGATGCAATTGTTTGGTGAAAACACTGACGT CGAAGGTATTGACACGCTTAATGCCTGTTACGGTGGTACCAACGCGTTGTTCAACTCTTTGAACTGGATTGAAT CTAACGCATGGGATGGTAGAGACGCCATTGTAGTTTGCGGTGATATTGCCATCTACGATAAGGGTGCCGCAAG ACCAACCGGTGGTGCCGGTACTGTTGCTATGTGGATCGGTCCTGATGCTCCAATTGTATTTGACTCTGTAAGAG CTTCTTACATGGAACACGCCTACGATTTTTACAAGCCAGATTTCACCAGCGAATATCCTTACGTCGATGGTCATT TTTCATTAACTTGTTACGTCAAGGCTCTTGATCAAGTTTACAAGAGTTATTCCAAGAAGGCTATTTCTAAAGGGTT GGTTAGCGATCCCGCTGGTTCGGATGCTTTGAACGTTTTGAAATATTTCGACTACAACGTTTTCCATGTTCCAAC CTGTAAATTGGTCACAAAATCATACGGTAGATTACTATATAACGATTTCAGAGCCAATCCTCAATTGTTCCCAGA AGTTGACGCCGAATTAGCTACTCGCGATTATGACGAATCTTTAACCGATAAGAACATTGAAAAAACTTTTGTTAA TGTTGCTAAGCCATTCCACAAAGAGAGAGTTGCCCAATCTTTGATTGTTCCAACAAACACAGGTAACATGTACAC CGCATCTGTTTATGCCGCCTTTGCATCTCTATTAAACTATGTTGGATCTGACGACTTACAAGGCAAGCGTGTTG GTTTATTTTCTTACGGTTCCGGTTTAGCTGCATCTCTATATTCTTGCAAAATTGTTGGTGACGTCCAACATATTAT CAAGGAATTAGATATTACTAACAAATTAGCCAAGAGAATCACCGAAACTCCAAAGGATTACGAAGCTGCCATCG AATTGAGAGAAAATGCCCATTTGAAGAAGAACTTCAAACCTCAAGGTTCCATTGAGCATTTGCAAAGTGGTGTTT ACTACTTGACCAACATCGATGACAAATTTAGAAGATCTTACGATGTTAAAAAATAA

Truncated HMG-CoA Reductase

ATGGTTGCGGTACGTAGGAAGGCTCTTTCAATTTTGGCAGAAGCTCCTGTATTAGCATCTGATCGTTTACCATAT AAAAATTATGACTACGACCGCGTATTTGGCGCTTGTTGTGAAAATGTTATAGGTTACATGCCTTTGCCCGTTGGT GTTATAGGCCCCTTGGTTATCGATGGTACATCTTATCATATACCAATGGCAACTACAGAGGGTTGTTTGGTAGCT TCTGCCATGCGTGGCTGTAAGGCAATCAATGCTGGCGGTGGTGCAACAACTGTTTTAACTAAGGATGGTATGA CAAGAGGCCCAGTAGTCCGTTTCCCAACTTTGAAAAGATCTGGTGCCTGTAAGATATGGTTAGACTCAGAAGAG GGACAAAACGCAATTAAAAAAGCTTTTAACTCTACATCAAGATTTGCACGTCTGCAACATATTCAAACTTGTCTA GCAGGAGATTTACTCTTCATGAGATTTAGAACAACTACTGGTGACGCAATGGGTATGAATATGATTTCTAAAGGT GTCGAATACTCATTAAAGCAAATGGTAGAAGAGTATGGCTGGGAAGATATGGAGGTTGTCTCCGTTTCTGGTAA CTACTGTACCGACAAAAAACCAGCTGCCATCAACTGGATCGAAGGTCGTGGTAAGAGTGTCGTCGCAGAAGCT ACTATTCCTGGTGATGTTGTCAGAAAAGTGTTAAAAAGTGATGTTTCCGCATTGGTTGAGTTGAACATTGCTAAG AATTTGGTTGGATCTGCAATGGCTGGGTCTGTTGGTGGATTTAACGCACATGCAGCTAATTTAGTGACAGCTGT TTTCTTGGCATTAGGACAAGATCCTGCACAAAATGTTGAAAGTTCCAACTGTATAACATTGATGAAAGAAGTGGA CGGTGATTTGAGAATTTCCGTATCCATGCCATCCATCGAAGTAGGTACCATCGGTGGTGGTACTGTTCTAGAAC CACAAGGTGCCATGTTGGACTTATTAGGTGTAAGAGGCCCGCATGCTACCGCTCCTGGTACCAACGCACGTCA ATTAGCAAGAATAGTTGCCTGTGCCGTCTTGGCAGGTGAATTATCCTTATGTGCTGCCCTAGCAGCCGGCCATT TGGTTCAAAGTCATATGACCCACAACAGG

Figure 12A (continued)

Mevalonate Kinase

ATGTCATTACCGTTCTTAACTTCTGCACCGGGAAAGGTTATTATTTTTGGTGAACACTCTGCTGTGTACAACAAG CCTGCCGTCGCTGCTAGTGTGTCTGCGTTGAGAACCTACCTGCTAATAAGCGAGTCATCTGCACCAGATACTAT TGAATTGGACTTCCCGGACATTAGCTTTAATCATAAGTGGTCCATCAATGATTTCAATGCCATCACCGAGGATCA AGTAAACTCCCAAAAATTGGCCAAGGCTCAACAAGCCACCGATGGCTTGTCTCAGGAACTCGTTAGTCTTTTGG ATCCGTTGTTAGCTCAACTATCCGAATCCTTCCACTACCATGCAGCGTTTTGTTTCCTGTATATGTTTGTTTGCCT ATGCCCCCATGCCAAGAATATTAAGTTTTCTTTAAAGTCTACTTTACCCATCGGTGCTGGGTTGGGCTCAAGCG CCTCTATTTCTGTATCACTGGCCTTAGCTATGGCCTACTTGGGGGGGTTAATAGGATCTAATGACTTGGAAAAG CTGTCAGAAAACGATAAGCATATAGTGAATCAATGGGCCTTCATAGGTGAAAAGTGTATTCACGGTACCCCTTC AGGAATAGATAACGCTGTGGCCACTTATGGTAATGCCCTGCTATTTGAAAAAGACTCACATAATGGAACAATAAA CACAAACAATTTTAAGTTCTTAGATGATTTCCCAGCCATTCCAATGATCCTAACCTATACTAGAATTCCAAGGTCT ACAAAAGATCTTGTTGCTCGCGTTCGTGTGTTGGTCACCGAGAAATTTCCTGAAGTTATGAAGCCAATTCTAGAT GCCATGGGTGAATGTGCCCTACAAGGCTTAGAGATCATGACTAAGTTAAGTAAATGTAAAGGCACCGATGACGA GGCTGTAGAAACTAATAATGAACTGTATGAACAACTATTGGAATTGATAAGAATAAATCATGGACTGCTTGTCTC AATCGGTGTTTCTCATCCTGGATTAGAACTTATTAAAAATCTGAGCGATGATTTGAGAATTGGCTCCACAAAACT TACCGGTGCTGGTGGCGGCGGTTGCTCTTTGACTTTGTTACGAAGAGACATTACTCAAGAGCAAATTGACAGCT TCAAAAAGAAATTGCAAGATGATTTTAGTTACGAGACATTTGAAACAGACTTGGGTGGGACTGGCTGCTGTTTG TTAAGCGCAAAAAATTTGAATAAAGATCTTAAAATCAAATCCCTAGTATTCCAATTATTTGAAAATAAAACTACCA CAAAGCAACAAATTGACGATCTATTATTGCCAGGAAACACGAATTTACCATGGACTTCATAA

Phosphomevalonate Kinase

ATGTCAGAGTTGAGAGCCTTCAGTGCCCCAGGGAAAGCGTTACTAGCTGGTGGATATTTAGTTTTAGATACAAA ATATGAAGCATTTGTAGTCGGATTATCGGCAAGAATGCATGCTGTAGCCCATCCTTACGGTTCATTGCAAGGGT CTGATAAGTTTGAAGTGCGTGTGAAAAGTAAACAATTTAAAGATGGGGAGTGGCTGTACCATATAAGTCCTAAA AGTGGCTTCATTCCTGTTTCGATAGGCGGATCTAAGAACCCTTTCATTGAAAAAGTTATCGCTAACGTATTTAGC TACTTTAAACCTAACATGGACGACTACTGCAATAGAAACTTGTTCGTTATTGATATTTTCTCTGATGATGCCTACC ATTCTCAGGAGGATAGCGTTACCGAACATCGTGGCAACAGAAGATTGAGTTTTCATTCGCACAGAATTGAAGAA GTTCCCAAAACAGGGCTGGGCTCCTCGGCAGGTTTAGTCACAGTTTTAACTACAGCTTTGGCCTCCTTTTTTGT ATCGGACCTGGAAAATAATGTAGACAAATATAGAGAAGTTATTCATAATTTAGCACAAGTTGCTCATTGTCAAGC TCAGGGTAAAATTGGAAGCGGGTTTGATGTAGCGGCGGCAGCATATGGATCTATCAGATATAGAAGATTCCCA CCCGCATTAATCTCTAATTTGCCAGATATTGGAAGTGCTACTTACGGCAGTAAACTGGCGCATTTGGTTGATGA AGAAGACTGGAATATTACGATTAAAAGTAACCATTTACCTTCGGGATTAACTTTATGGATGGGCGATATTAAGAA TGGTTCAGAAACAGTAAAACTGGTCCAGAAGGTAAAAAATTGGTATGATTCGCATATGCCAGAAAGCTTGAAAA TATATACAGAACTCGATCATGCAAATTCTAGATTTATGGATGGACTATCTAAACTAGATCGCTTACACGAGACTC ATGACGATTACAGCGATCAGATATTTGAGTCTCTTGAGAGGAATGACTGTACCTGTCAAAAGTATCCTGAAATCA CAGAAGTTAGAGATGCAGTTGCCACAATTAGACGTTCCTTTAGAAAAATAACTAAAGAATCTGGTGCCGATATC GAACCTCCCGTACAAACTAGCTTATTGGATGATTGCCAGACCTTAAAAGGAGTTCTTACTTGCTTAATACCTGGT GCTGGTGGTTATGACGCCATTGCAGTGATTACTAAGCAAGATGTTGATCTTAGGGCTCAAACCGCTAATGACAA AAGATTTTCTAAGGTTCAATGGCTGGATGTAACTCAGGCTGACTGGGGTGTTAGGAAAGAAAAAGATCCGGAAA CTTATCTTGATAAATAA

Figure 12A (continued)

Diphosphomevalonate Decarboxylase

ATGACCGTTTACACAGCATCCGTTACCGCACCCGTCAACATCGCAACCCTTAAGTATTGGGGGAAAAGGGACA CGAAGTTGAATCTGCCCACCAATTCGTCCATATCAGTGACTTTATCGCAAGATGACCTCAGAACGTTGACCTCT GCGGCTACTGCACCTGAGTTTGAACGCGACACTTTGTGGTTAAATGGAGAACCACACAGCATCGACAATGAAA GAACTCAAAATTGTCTGCGCGACCTACGCCAATTAAGAAAGGAAATGGAATCGAAGGACGCCTCATTGCCCAC ATTATCTCAATGGAAACTCCACATTGTCTCCGAAAATAACTTTCCTACAGCAGCTGGTTTAGCTTCCTCCGCTGC TGGCTTTGCTGCATTGGTCTCTGCAATTGCTAAGTTATACCAATTACCACAGTCAACTTCAGAAATATCTAGAAT AGCAAGAAAGGGGTCTGGTTCAGCTTGTAGATCGTTGTTTGGCGGATACGTGGCCTGGGAAATGGGAAAAGCT GAAGATGGTCATGATTCCATGGCAGTACAAATCGCAGACAGCTCTGACTGGCCTCAGATGAAAGCTTGTGTCCT AGTTGTCAGCGATATTAAAAAGGATGTGAGTTCCACTCAGGGTATGCAATTGACCGTGGCAACCTCCGAACTAT TTAAAGAAAGAATTGAACATGTCGTACCAAAGAGATTTGAAGTCATGCGTAAAGCCATTGTTGAAAAAGATTTCG CCACCTTTGCAAAGGAAACAATGATGGATTCCAACTCTTTCCATGCCACATGTTTGGACTCTTTCCCTCCAATAT TCTACATGAATGACACTTCCAAGCGTATCATCAGTTGGTGCCACACCATTAATCAGTTTTACGGAGAAACAATCG TTGCATACACGTTTGATGCAGGTCCAAATGCTGTGTTGTACTACTTAGCTGAAAATGAGTCGAAACTCTTTGCAT TTATCTATAAATTGTTTGGCTCTGTTCCTGGATGGGACAAGAAATTTACTACTGAGCAGCTTGAGGCTTTCAACC ATCAATTTGAATCATCTAACTTTACTGCACGTGAATTGGATCTTGAGTTGCAAAAGGATGTTGCCAGAGTGATTT TAACTCAAGTCGGTTCAGGCCCACAAGAAACAAACGAATCTTTGATTGACGCAAAGACTGGTCTACCAAAGGAA TAA

Isopentenyl-Diphosphate Delta-lsomerase

ATGACTGCCGACAACAATAGTATGCCCCATGGTGCAGTATCTAGTTACGCCAAATTAGTGCAAAACCAAACACC TGAAGACATTTTGGAAGAGTTTCCTGAAATTATTCCATTACAACAAAGACCTAATACCCGATCTAGTGAGACGTC AAATGACGAAAGCGGAGAAACATGTTTTTCTGGTCATGATGAGGAGCAAATTAAGTTAATGAATGAAAATTGTAT TGTTTTGGATTGGGACGATAATGCTATTGGTGCCGGTACCAAGAAAGTTTGTCATTTAATGGAAAATATTGAAAA GGGTTTACTACATCGTGCATTCTCCGTCTTTATTTTCAATGAACAAGGTGAATTACTTTTACAACAAAGAGCCAC TGAAAAAATAACTTTCCCTGATCTTTGGACTAACACATGCTGCTCTCATCCACTATGTATTGATGACGAATTAGG TTTGAAGGGTAAGCTAGACGATAAGATTAAGGGCGCTATTACTGCGGCGGTGAGAAAACTAGATCATGAATTAG GTATTCCAGAAGATGAAACTAAGACAAGGGGTAAGTTTCACTTTTTAAACAGAATCCATTACATGGCACCAAGCA ATGAACCATGGGGTGAACATGAAATTGATTACATCCTATTTTATAAGATCAACGCTAAAGAAAACTTGACTGTCA ACCCAAACGTCAATGAAGTTAGAGACTTCAAATGGGTTTCACCAAATGATTTGAAAACTATGTTTGCTGACCCAA GTTACAAGTTTACGCCTTGGTTTAAGATTATTTGCGAGAATTACTTATTCAACTGGTGGGAGCAATTAGATGACC TTTCTGAAGTGGAAAATGACAGGCAAATTCATAGAATGCTATAA

Geranyl-Diphosphate Synthase (ERG20ww)

ATGGCTTCAGAAAAGGAAATAAGAAGAGAAAGATTCTTGAACGTATTCCCAAAGTTAGTTGAAGAATTGAACGCT AGTTTGTTAGCTTATGGTATGCCTAAAGAAGCCTGCGATTGGTATGCTCACTCTTTAAACTACAATACTCCAGGT GGTAAATTGAATAGAGGTTTGAGTGTAGTTGATACTTATGCTATCTTGTCTAACAAAACCGTTGAACAATTAGGT CAAGAAGAATACGAAAAGGTCGCTATCTTGGGTTGGTGTATTGAATTGTTGCAAGCATACTTTTTGGTTGCCGAT GACATGATGGATAAGTCTATAACAAGAAGAGGTCAACCATGCTGGTACAAAGTTCCAGAAGTTGGTGAAATAGC CATAAATGATGCTTTTATGTTGGAAGCCGCTATCTATAAATTGTTGAAGTCACATTTCAGAAACGAAAAGTACTA CATCGATATTACCGAATTATTCCACGAAGTTACTTTCCAAACAGAATTGGGTCAATTGATGGATTTGATAACTGC ACCTGAAGATAAAGTTGACTTGTCAAAGTTTTCCTTGAAGAAACATTCATTCATCGTCACCTTTGAAACTGCTTAT TACTCCTTCTATTTGCCAGTCGCCTTGGCTATGTACGTAGCTGGTATTACTGATGAAAAAGACTTGAAGCAAGCA AGAGATGTTTTGATACCTTTGGGTGAATACTTCCAAATCCAAGATGACTACTTAGACTGTTTCGGTACTCCAGAA CAAATAGGTAAAATCGGTACAGATATTCAAGACAATAAGTGCAGTTGGGTTATTAACAAGGCTTTGGAATTAGCA TCTGCCGAACAAAGAAAGACTTTGGATGAAAACTACGGTAAAAAGGACTCAGTTGCTGAAGCAAAGTGTAAGAA AATTTTTAATGATTTGAAGATTGAACAATTGTACCATGAATACGAAGAATCCATCGCTAAAGACTTAAAGGCAAA GATTAGTCAAGTTGATGAATCAAGAGGTTTTAAAGCCGACGTTTTGACAGCTTTCTTGAATAAGGTCTACAAGAG ATCAAAGTAG

Figure 12A (continued)

Olivetol Synthase

ATGAATCATTTGAGAGCTGAAGGTCCAGCTTCTGTTTTGGCTATTGGTACTGCTAATCCAGAAAATATTTTGTTG CAAGATGAATTTCCAGATTATTATTTTAGAGTTACTAAATCTGAACATATGACTCAATTGAAAGAAAAATTTAGAA AAATTTGTGATAAATCTATGATTAGAAAAAGAAATTGTTTTTTGAATGAAGAACATTTGAAACAAAATCCAAGATT GGTTGAACATGAAATGCAAACTTTGGATGCTAGACAAGATATGTTGGTTGTTGAAGTTCCAAAATTGGGTAAAGA TGCTTGTGCTAAAGCTATTAAAGAATGGGGTCAACCAAAATCTAAAATTACTCATTTGATTTTTACTTCTGCTTCT ACTACTGATATGCCAGGTGCTGATTATCATTGTGCTAAATTGTTGGGTTTGTCTCCATCTGTTAAAAGAGTTATG ATGTATCAATTGGGTTGTTATGGTGGTGGTACTGTTTTGAGAATTGCTAAAGATATTGCTGAAAATAATAAAGGT GCTAGAGTTTTGGCTGTTTGTTGTGATATTATGGCTTGTTTGTTTAGAGGTCCATCTGAATCTGATTTGGAATTG TTGGTTGGTCAAGCTATTTTTGGTGATGGTGCTGCTGCTGTTATTGTTGGTGCTGAACCAGATGAATCTGTTGG TGAAAGACCAATTTTTGAATTGGTTTCTACTGGTCAAACTATTTTGCCAAATTCTGAAGGTACTATTGGTGGTCAT ATTAGAGAAGCTGGTTTGATTTTTGATTTGCATAAAGATGTTCCAATGTTGATTTCTAATAATATTGAAAAATGTTT GATTGAAGCTTTTACTCCAATTGGTATTTCTGATTGGAATTCTATTTTTTGGATTACTCATCCAGGTGGTAAAGCT ATTTTGGATAAAGTTGAAGAAAAATTGCATTTGAAATCTGATAAATTTGTTGATTCTAGACATGTTTTGTCTGAAC ATGGTAATATGTCTTCTTCTACTGTTTTGTTTGTTATGGATGAATTGAGAAAAAGATCTTTGGAAGAAGGTAAATC TACTACTGGTGATGGTTTTGAATGGGGTGTTTTGTTTGGTTTTGGTCCAGGTTTGACTGTTGAAAGAGTTGTTGT TAGATCTGTTCCAATTAAATAT

Olivetolic Acid Cyclase

ATGGCTGTTAAACATTTGATTGTTTTGAAATTTAAAGATGAAATTACTGAAGCTCAAAAAGAAGAATTTTTTAAAA CTTATGTTAATTTGGTTAATATTATTCCAGCTATGAAAGATGTTTATTGGGGTAAAGATGTTACTCAAAAAAATAA AGAAGAAGGTTATACTCATATTGTTGAAGTTACTTTTGAATCTGTTGAAACTATTCAAGATTATATTATTCATCCA GCTCATGTTGGTTTTGGTGATGTTTATAGATCTTTTTGGGAAAAATTGTTGATTTTTGATTATACTCCAAGAAAA

CBGA Synthase

ATGGGTTTGTCTTCTGTTTGTACTTTTTCTTTTCAAACTAATTATCATACTTTGTTGAATCCACATAATAATAATCC AAAAACTTCTTTGTTGTGTTATAGACATCCAAAAACTCCAATTAAATATTCTTATAATAATTTTCCATCTAAACATT GTTCTACTAAATCTTTTCATTTGCAAAATAAATGTTCTGAATCTTTGTCTATTGCTAAAAATTCTATTAGAGCTGCT ACTACTAATCAAACTGAACCACCAGAATCTGATAATCATTCTGTTGCTACTAAAATTTTGAATTTTGGTAAAGCTT GTTGGAAATTGCAAAGACCATATACTATTATTGCTTTTACTTCTTGTGCTTGTGGTTTGTTTGGTAAAGAATTGTT GCATAATACTAATTTGATTTCTTGGTCTTTGATGTTTAAAGCTTTTTTTTTTTTGGTTGCTATTTTGTGTATTGCTT CTTTTACTACTACTATTAATCAAATTTATGATTTGCATATTGATAGAATTAATAAACCAGATTTGCCATTGGCTTCT GGTGAAATTTCTGTTAATACTGCTTGGATTATGTCTATTATTGTTGCTTTGTTTGGTTTGATTATTACTATTAAAAT GAAAGGTGGTCCATTGTATATTTTTGGTTATTGTTTTGGTATTTTTGGTGGTATTGTTTATTCTGTTCCACCATTTA GATGGAAACAAAATCCATCTACTGCTTTTTTGTTGAATTTTTTGGCTCATATTATTACTAATTTTACTTTTTATTAT GCTTCTAGAGCTGCTTTGGGTTTGCCATTTGAATTGAGACCATCTTTTACTTTTTTGTTGGCTTTTATGAAATCTA TGGGTTCTGCTTTGGCTTTGATTAAAGATGCTTCTGATGTTGAAGGTGATACTAAATTTGGTATTTCTACTTTGG CTTCTAAATATGGTTCTAGAAATTTGACTTTGTTTTGTTCTGGTATTGTTTTGTTGTCTTATGTTGCTGCTATTTTG GCTGGTATTATTTGGCCACAAGCTTTTAATTCTAATGTTATGTTGTTGTCTCATGCTATTTTGGCTTTTTGGTTGA TTTTGCAAACTAGAGATTTTGCTTTGACTAATTATGATCCAGAAGCTGGTAGAAGATTTTATGAATTTATGTGGAA ATTGTATTATGCTGAATATTTGGTTTATGTTTTTATT

Figure 12A (continued)

AcetvI-CoA Carboxylase

ATGAGCGAAGAAAGCTTATTCGAGTCTTCTCCACAGAAGATGGAGTACGAAATTACAAACTACTCAGAAAGACA TACAGAACTTCCAGGTCATTTCATTGGCCTCAATACAGTAGATAAACTAGAGGAGTCCCCGTTAAGGGACTTTG TTAAGAGTCACGGTGGTCACACGGTCATATCCAAGATCCTGATAGCAAATAATGGTATTGCCGCCGTGAAAGAA ATTAGATCCGTCAGAAAATGGGCATACGAGACGTTCGGCGATGACAGAACCGTCCAATTCGTCGCCATGGCCA CCCCAGAAGATCTGGAGGCCAACGCAGAATATATCCGTATGGCCGATCAATACATTGAAGTGCCAGGTGGTAC TAATAATAACAACTACGCTAACGTAGACTTGATCGTAGACATCGCCGAAAGAGCAGACGTAGACGCCGTATGG GCTGGCTGGGGTCACGCCTCCGAGAATCCACTATTGCCTGAAAAATTGTCCCAGTCTAAGAGGAAAGTCATCTT TATTGGGCCTCCAGGTAACGCCATGAGGTCTTTAGGTGATAAAATCTCCTCTACCATTGTCGCTCAAAGTGCTA AAGTCCCATGTATTCCATGGTCTGGTACCGGTGTTGACACCGTTCACGTGGACGAGAAAACCGGTCTGGTCTC TGTCGACGATGACATCTATCAAAAGGGTTGTTGTACCTCTCCTGAAGATGGTTTACAAAAGGCCAAGCGTATTG GTTTTCCTGTCATGATTAAGGCATCCGAAGGTGGTGGTGGTAAAGGTATCAGACAAGTTGAACGTGAAGAAGAT TTCATCGCTTTATACCACCAGGCAGCCAACGAAATTCCAGGCTCCCCCATTTTCATCATGAAGTTGGCCGGTAG AGCGCGTCACTTGGAAGTTCAACTGCTAGCAGATCAGTACGGTACAAATATTTCCTTGTTCGGTAGAGACTGTT CCGTTCAGAGACGTCATCAAAAAATTATCGAAGAAGCACCAGTTACAATTGCCAAGGCTGAAACATTTCACGAG ATGGAAAAGGCTGCCGTCAGACTGGGGAAACTAGTCGGTTATGTCTCTGCCGGTACCGTGGAGTATCTATATT CTCATGATGATGGAAAATTCTACTTTTTAGAATTGAACCCAAGATTACAAGTCGAGCATCCAACAACGGAAATGG TCTCCGGTGTTAACTTACCTGCAGCTCAATTACAAATCGCTATGGGTATCCCTATGCATAGAATAAGTGACATTA GAACTTTATATGGTATGAATCCTCATTCTGCCTCAGAAATCGATTTCGAATTCAAAACTCAAGATGCCACCAAGA AACAAAGAAGACCTATTCCAAAGGGTCATTGTACCGCTTGTCGTATCACATCAGAAGATCCAAACGATGGATTC AAGCCATCGGGTGGTACTTTGCATGAACTAAACTTCCGTTCTTCCTCTAATGTTTGGGGTTACTTCTCCGTGGG TAACAATGGTAATATTCACTCCTTTTCGGACTCTCAGTTCGGCCATATTTTTGCTTTTGGTGAAAATAGACAAGCT TCCAGGAAACACATGGTTGTTGCCCTGAAGGAATTGTCCATTAGGGGTGATTTCAGAACTACTGTGGAATACTT GATCAAACTTTTGGAAACTGAAGATTTCGAGGATAACACTATTACCACCGGTTGGTTGGACGATTTGATTACTCA TAAAATGACCGCTGAAAAGCCTGATCCAACTCTTGCCGTCATTTGCGGTGCCGCTACAAAGGCTTTCTTAGCAT CTGAAGAAGCCCGCCACAAGTATATCGAATCCTTACAAAAGGGACAAGTTCTATCTAAAGACCTACTGCAAACT ATGTTCCCTGTAGATTTTATCCATGAGGGTAAAAGATACAAGTTCACCGTAGCTAAATCCGGTAATGACCGTTAC ACATTATTTATCAATGGTTCTAAATGTGATATCATACTGCGTCAACTATCTGATGGTGGTCTTTTGATTGCCATAG GCGGTAAATCGCATACCATCTATTGGAAAGAAGAAGTTGCTGCTACAAGATTATCCGTTGACTCTATGACTACTT TGTTGGAAGTTGAAAACGATCCAACCCAGTTGCGTACTCCATCCCCTGGTAAATTGGTTAAATTCTTGGTGGAA AATGGTGAACACATTATCAAGGGCCAACCATATGCAGAAATTGAAGTTATGAAAATGCAAATGCCTTTGGTTTCT CAAGAAAATGGTATCGTCCAGTTATTAAAGCAACCTGGTTCTACCATTGTTGCAGGTGATATCATGGCTATTATG ACTCTTGACGATCCATCCAAGGTCAAGCACGCTCTACCATTTGAAGGTATGCTGCCAGATTTTGGTTCTCCAGT TATCGAAGGAACCAAACCTGCCTATAAATTCAAGTCATTAGTGTCTACTTTGGAAAACATTTTGAAGGGTTATGA CAACCAAGTTATTATGAACGCTTCCTTGCAACAATTGATAGAGGTTTTGAGAAATCCAAAACTGCCTTACTCAGA ATGGAAACTACACATCTCTGCTTTACATTCAAGATTGCCTGCTAAGCTAGATGAACAAATGGAAGAGTTAGTTGC ACGTTCTTTGAGACGTGGTGCTGTTTTCCCAGCTAGACAATTAAGTAAATTGATTGATATGGCCGTGAAGAATCC TGAATACAACCCCGACAAATTGCTGGGCGCCGTCGTGGAACCATTGGCGGATATTGCTCATAAGTACTCTAAC GGGTTAGAAGCCCATGAACATTCTATATTTGTCCATTTCTTGGAAGAATATTACGAAGTTGAAAAGTTATTCAAT GGTCCAAATGTTCGTGAGGAAAATATCATTCTGAAATTGCGTGATGAAAACCCTAAAGATCTAGATAAAGTTGCG CTAACTGTTTTGTCTCATTCGAAAGTTTCAGCGAAGAATAACCTGATCCTAGCTATCTTGAAACATTATCAACCAT TGTGCAAGTTATCTTCTAAAGTTTCTGCCATTTTCTCTACTCCTCTACAACATATTGTTGAACTAGAATCTAAGGC TACCGCTAAGGTCGCTCTACAAGCAAGAGAAATTTTGATTCAAGGCGCTTTACCTTCGGTCAAGGAAAGAACTG AACAAATTGAACATATCTTAAAATCCTCTGTTGTGAAGGTTGCCTATGGCTCATCCAATCCAAAGCGCTCTGAAC CAGATTTGAATATCTTGAAGGACTTGATCGATTCTAATTACGTTGTGTTCGATGTTTTACTTCAATTCCTAACCCA TCAAGACCCAGTTGTGACTGCTGCAGCTGCTCAAGTCTATATTCGTCGTGCTTATCGTGCTTACACCATAGGAG ATATTAGAGTTCACGAAGGTGTCACAGTTCCAATTGTTGAATGGAAATTCCAACTACCTTCAGCTGCGTTCTCCA CCTTTCCAACTGTTAAATCTAAAATGGGTATGAACAGGGCTGTTTCTGTTTCAGATTTGTCATATGTTGCAAACA GTCAGTCATCTCCGTTAAGAGAAGGTATTTTGATGGCTGTGGATCATTTAGATGATGTTGATGAAATTTTGTCAC AAAGTTTGGAAGTTATTCCTCGTCACCAATCTTCTTCTAACGGACCTGCTCCTGATCGTTCTGGTAGCTCCGCAT CGTTGAGTAATGTTGCTAATGTTTGTGTTGCTTCTACAGAAGGTTTCGAATCTGAAGAGGAAATTTTGGTAAGGT TGAGAGAAATTTTGGATTTGAATAAGCAGGAATTAATCAATGCTTCTATCCGTCGTATCACATTTATGTTCGGTTT TAAAGATGGGTCTTATCCAAAGTATTATACTTTTAACGGTCCAAATTATAACGAAAATGAAACAATTCGTCACATT

Figure 12A (continued)

GAGCCGGCTTTGGCCTTCCAACTGGAATTAGGAAGATTGTCCAACTTCAACATTAAACCAATTTTCACTGATAAT AGAAACATCCATGTCTACGAAGCTGTTAGTAAGACTTCTCCATTGGATAAGAGATTCTTTACAAGAGGTATTATT AGAACGGGTCATATCCGTGATGACATTTCTATTCAAGAATATCTGACTTCTGAAGCTAACAGATTGATGAGTGAT ATATTGGATAATTTAGAAGTCACCGACACTTCAAATTCTGATTTGAATCATATCTTCATCAACTTCATTGCGGTGT TTGATATCTCTCCAGAAGATGTCGAAGCCGCCTTCGGTGGTTTCTTAGAAAGATTTGGTAAGAGATTGTTGAGA TTGCGTGTTTCTTCTGCCGAAATTAGAATCATCATCAAAGATCCTCAAACAGGTGCCCCAGTACCATTGCGTGC CTTGATCAATAACGTTTCTGGTTATGTTATCAAAACAGAAATGTACACCGAAGTCAAGAACGCAAAAGGTGAATG GGTATTTAAGTCTTTGGGTAAACCTGGATCCATGCATTTAAGACCTATTGCTACTCCTTACCCTGTTAAGGAATG GTTGCAACCAAAACGTTATAAGGCACACTTGATGGGTACCACATATGTCTATGACTTCCCAGAATTATTCCGCCA AGCATCGTCATCCCAATGGAAAAATTTCTCTGCAGATGTTAAGTTAACAGATGATTTCTTTATTTCCAACGAGTT GATTGAAGATGAAAACGGCGAATTAACTGAGGTGGAAAGAGAACCTGGTGCCAACGCTATTGGTATGGTTGCC TTTAAGATTACTGTAAAGACTCCTGAATATCCAAGAGGCCGTCAATTTGTTGTTGTTGCTAACGATATCACATTC AAGATCGGTTCCTTTGGTCCACAAGAAGACGAATTCTTCAATAAGGTTACTGAATATGCTAGAAAGCGTGGTAT CCCAAGAATTTACTTGGCTGCAAACTCAGGTGCCAGAATTGGTATGGCTGAAGAGATTGTTCCACTATTTCAAG TTGCATGGAATGATGCTGCCAATCCGGACAAGGGCTTCCAATACTTATACTTAACAAGTGAAGGTATGGAAACT TTAAAGAAATTTGACAAAGAAAATTCTGTTCTCACTGAACGTACTGTTATAAACGGTGAAGAAAGATTTGTCATCA AGACAATTATTGGTTCTGAAGATGGGTTAGGTGTCGAATGTCTACGTGGATCTGGTTTAATTGCTGGTGCAACG TCAAGGGCTTACCACGATATCTTCACTATCACCTTAGTCACTTGTAGATCCGTCGGTATCGGTGCTTATTTGGTT CGTTTGGGTCAAAGAGCTATTCAGGTCGAAGGCCAGCCAATTATTTTAACTGGTGCTCCTGCAATCAACAAAAT GCTGGGTAGAGAAGTTTATACTTCTAACTTACAATTGGGTGGTACTCAAATCATGTATAACAACGGTGTTTCACA TTTGACTGCTGTTGACGATTTAGCTGGTGTAGAGAAGATTGTTGAATGGATGTCTTATGTTCCAGCCAAGCGTA ATATGCCAGTTCCTATCTTGGAAACTAAAGACACATGGGATAGACCAGTTGATTTCACTCCAACTAATGATGAAA CTTACGATGTAAGATGGATGATTGAAGGTCGTGAGACTGAAAGTGGATTTGAATATGGTTTGTTTGATAAAGGG TCTTTCTTTGAAACTTTGTCAGGATGGGCCAAAGGTGTTGTCGTTGGTAGAGCCCGTCTTGGTGGTATTCCACT GGGTGTTATTGGTGTTGAAACAAGAACTGTCGAGAACTTGATTCCTGCTGATCCAGCTAATCCAAATAGTGCTG AAACATTAATTCAAGAACCTGGTCAAGTTTGGCATCCAAACTCCGCCTTCAAGACTGCTCAAGCTATCAATGACT TTAACAACGGTGAACAATTGCCAATGATGATTTTGGCCAACTGGAGAGGTTTCTCTGGTGGTCAACGTGATATG TTCAACGAAGTCTTGAAGTATGGTTCGTTTATTGTTGACGCATTGGTGGATTACAAACAACCAATTATTATCTATA TCCCACCTACCGGTGAACTAAGAGGTGGTTCATGGGTTGTTGTCGATCCAACTATCAACGCTGACCAAATGGAA ATGTATGCCGACGTCAACGCTAGAGCTGGTGTTTTGGAACCACAAGGTATGGTTGGTATCAAGTTCCGTAGAGA AAAATTGCTGGACACCATGAACAGATTGGATGACAAGTACAGAGAATTGAGATCTCAATTATCCAACAAGAGTTT GGCTCCAGAAGTACATCAGCAAATATCCAAGCAATTAGCTGATCGTGAGAGAGAACTATTGCCAATTTACGGAC AAATCAGTCTTCAATTTGCTGATTTGCACGATAGGTCTTCACGTATGGTGGCCAAGGGTGTTATTTCTAAGGAAC TGGAATGGACCGAGGCACGTCGTTTCTTCTTCTGGAGATTGAGAAGAAGATTGAACGAAGAATATTTGATTAAA AGGTTGAGCCATCAGGTAGGCGAAGCATCAAGATTAGAAAAGATCGCAAGAATTAGATCGTGGTACCCTGCTT CAGTGGACCATGAAGATGATAGGCAAGTCGCAACATGGATTGAAGAAAACTACAAAACTTTGGACGATAAACTA AAGGGTTTGAAATTAGAGTCATTCGCTCAAGACTTAGCTAAAAAGATCAGAAGCGACCATGACAATGCTATTGAT GGATTATCTGAAGTTATCAAGATGTTATCTACCGATGATAAAGAAAAATTGTTGAAGACTTTGAAATAA

CBDA Synthase

ATGAAATGTTCTACTTTTTCTTTTTGGTTTGTTTGTAAAATTATTTTTTTTTTTTTTTCTTTTAATATTCAAACTTCTA TTGCTAATCCAAGAGAAAATTTTTTGAAATGTTTTTCTCAATATATTCCAAATAATGCTACTAATTTGAAATTGGTT TATACTCAAAATAATCCATTGTATATGTCTGTTTTGAATTCTACTATTCATAATTTGAGATTTACTTCTGATACTAC TCCAAAACCATTGGTTATTGTTACTCCATCTCATGTTTCTCATATTCAAGGTACTATTTTGTGTTCTAAAAAAGTT GGTTTGCAAATTAGAACTAGATCTGGTGGTCATGATTCTGAAGGTATGTCTTATATTTCTCAAGTTCCATTTGTTA TTGTTGATTTGAGAAATATGAGATCTATTAAAATTGATGTTCATTCTCAAACTGCTTGGGTTGAAGCTGGTGCTA CTTTGGGTGAAGTTTATTATTGGGTTAATGAAAAAAATGAAAATTTGTCTTTGGCTGCTGGTTATTGTCCAACTGT TTGTGCTGGTGGTCATTTTGGTGGTGGTGGTTATGGTCCATTGATGAGAAATTATGGTTTGGCTGCTGATAATAT TATTGATGCTCATTTGGTTAATGTTCATGGTAAAGTTTTGGATAGAAAATCTATGGGTGAAGATTTGTTTTGGGCT TTGAGAGGTGGTGGTGCTGAATCTTTTGGTATTATTGTTGCTTGGAAAATTAGATTGGTTGCTGTTCCAAAATCT

Figure 12A (continued)

ACTATGTTTTCTGTTAAAAAAATTATGGAAATTCATGAATTGGTTAAATTGGTTAATAAATGGCAAAATATTGCTTA TAAATATGATAAAGATTTGTTGTTGATGACTCATTTTATTACTAGAAATATTACTGATAATCAAGGTAAAAATAAAA CTGCTATTCATACTTATTTTTCTTCTGTTTTTTTGGGTGGTGTTGATTCTTTGGTTGATTTGATGAATAAATCTTTT CCAGAATTGGGTATTAAAAAAACTGATTGTAGACAATTGTCTTGGATTGATACTATTATTTTTTATTCTGGTGTTG TTAATTATGATACTGATAATTTTAATAAAGAAATTTTGTTGGATAGATCTGCTGGTCAAAATGGTGCTTTTAAAATT AAATTGGATTATGTTAAAAAACCAATTCCAGAATCTGTTTTTGTTCAAATTTTGGAAAAATTGTATGAAGAAGATA TTGGTGCTGGTATGTATGCTTTGTATCCATATGGTGGTATTATGGATGAAATTTCTGAATCTGCTATTCCATTTCC ACATAGAGCTGGTATTTTGTATGAATTGTGGTATATTTGTTCTTGGGAAAAACAAGAAGATAATGAAAAACATTTG AATTGGATTAGAAATATTTATAATTTTATGACTCCATATGTTTCTAAAAATCCAAGATTGGCTTATTTGAATTATAG AGATTTGGATATTGGTATTAATGATCCAAAAAATCCAAATAATTATACTCAAGCTAGAATTTGGGGTGAAAAATAT TTTGGTAAAAATTTTGATAGATTGGTTAAAGTTAAAACTTTGGTTGATCCAAATAATTTTTTTAGAAATGAACAATC TATTCCACCATTGCCAAGACATAGACAT

CBCA Synthase

ATGAATTGTAGTACTTTCTCTTTCTGGTTTGTTTGTAAGATTATATTTTTTTTTCTTAGTTTCAATATACAAATTTCA ATTGCAAACCCTCAAGAAAATTTCCTTAAGTGCTTTTCAGAATATATCCCTAATAATCCTGCAAACCCTAAATTCA TTTATACACAACATGATCAGTTATATATGTCTGTCTTAAACTCTACCATTCAAAATTTGAGGTTCACGTCTGATAC AACCCCAAAGCCTTTAGTTATCGTGACACCCTCTAACGTTAGTCATATTCAGGCTAGTATCTTATGTTCAAAAAA AGTGGGTTTACAAATCAGAACTAGGTCTGGTGGTCATGACGCGGAAGGTCTGTCTTACATATCTCAGGTGCCGT TTGCAATCGTTGATCTACGTAATATGCATACAGTTAAAGTCGATATTCACTCTCAAACTGCATGGGTCGAGGCTG GTGCCACTCTAGGTGAAGTTTATTACTGGATCAATGAAATGAACGAGAATTTTTCCTTCCCAGGTGGTTATTGTC CTACTGTGGGTGTAGGCGGACACTTTTCTGGCGGGGGGTATGGTGCTTTGATGAGGAACTATGGTTTGGCCGC CGATAATATAATTGACGCCCATCTTGTAAACGTCGACGGGAAGGTTCTGGACCGTAAATCTATGGGTGAAGATT TATTCTGGGCGATAAGAGGTGGCGGGGGAGAGAACTTTGGTATTATCGCAGCTTGTAAGATTAAGTTAGTTGTT GTCCCCTCAAAAGCAACAATTTTTTCAGTGAAGAAGAACATGGAAATCCACGGTTTGGTAAAACTGTTTAATAAA TGGCAGAATATTGCCTACAAATACGATAAGGATTTGATGTTGACAACACATTTCAGAACTAGAAATATTACTGAC AACCACGGAAAGAACAAGACAACCGTCCATGGATATTTTAGTTCTATTTTCTTAGGCGGAGTTGATTCACTAGTA GACTTAATGAACAAGTCTTTCCCCGAATTGGGAATAAAAAAAACCGATTGCAAGGAATTATCCTGGATAGATACA ACAATATTCTACTCTGGAGTCGTTAATTATAATACGGCCAACTTTAAGAAGGAAATATTATTAGATCGTTCCGCA GGTAAAAAGACAGCTTTTTCCATAAAATTGGACTACGTCAAAAAATTAATTCCTGAGACAGCCATGGTAAAAATA TTGGAAAAATTGTACGAAGAGGAGGTAGGCGTGGGTATGTATGTGTTATACCCATACGGTGGTATTATGGATGA AATTTCTGAGAGCGCTATTCCCTTCCCCCATCGTGCAGGTATAATGTATGAATTATGGTACACAGCAACATGGG AAAAACAAGAGGATAACGAAAAGCATATTAATTGGGTACGTAGTGTGTACAACTTTACGACACCTTACGTGTCC CAAAATCCAAGATTAGCGTATTTGAACTATAGAGACTTAGATTTAGGTAAAACAAACCCTGAGTCTCCAAATAAT TACACCCAAGCCAGGATTTGGGGTGAAAAATACTTCGGCAAAAATTTCAATAGATTGGTTAAGGTAAAAACTAAG GCGGATCCAAACAATTTTTTTAGAAATGAGCAGAGTATTCCGCCCCTGCCTCCAAGACACCAT

Hexanoyl-CoA Synthetase

ATGGGTAAAAATTATAAATCTTTGGATTCTGTTGTTGCTTCTGATTTTATTGCTTTGGGTATTACTTCTGAAGTTG CTGAAACTTTGCATGGTAGATTGGCTGAAATTGTTTGTAATTATGGTGCTGCTACTCCACAAACTTGGATTAATA TTGCTAATCATATTTTGTCTCCAGATTTGCCATTTTCTTTGCATCAAATGTTGTTTTATGGTTGTTATAAAGATTTT GGTCCAGCTCCACCAGCTTGGATTCCAGATCCAGAAAAAGTTAAATCTACTAATTTGGGTGCTTTGTTGGAAAA AAGAGGTAAAGAATTTTTGGGTGTTAAATATAAAGATCCAATTTCTTCTTTTTCTCATTTTCAAGAATTTTCTGTTA GAAATCCAGAAGTTTATTGGAGAACTGTTTTGATGGATGAAATGAAAATTTCTTTTTCTAAAGATCCAGAATGTAT TTTGAGAAGAGATGATATTAATAATCCAGGTGGTTCTGAATGGTTGCCAGGTGGTTATTTGAATTCTGCTAAAAA TTGTTTGAATGTTAATTCTAATAAAAAATTGAATGATACTATGATTGTTTGGAGAGATGAAGGTAATGATGATTTG CCATTGAATAAATTGACTTTGGATCAATTGAGAAAAAGAGTTTGGTTGGTTGGTTATGCTTTGGAAGAAATGGGT TTGGAAAAAGGTTGTGCTATTGCTATTGATATGCCAATGCATGTTGATGCTGTTGTTATTTATTTGGCTATTGTTT

Figure 12A (continued)

TGGCTGGTTATGTTGTTGTTTCTATTGCTGATTCTTTTTCTGCTCCAGAAATTTCTACTAGATTGAGATTGTCTAA AGCTAAAGCTATTTTTACTCAAGATCATATTATTAGAGGTAAAAAAAGAATTCCATTGTATTCTAGAGTTGTTGAA GCTAAATCTCCAATGGCTATTGTTATTCCATGTTCTGGTTCTAATATTGGTGCTGAATTGAGAGATGGTGATATTT CTTGGGATTATTTTTTGGAAAGAGCTAAAGAATTTAAAAATTGTGAATTTACTGCTAGAGAACAACCAGTTGATG CTTATACTAATATTTTGTTTTCTTCTGGTACTACTGGTGAACCAAAAGCTATTCCATGGACTCAAGCTACTCCATT GAAAGCTGCTGCTGATGGTTGGTCTCATTTGGATATTAGAAAAGGTGATGTTATTGTTTGGCCAACTAATTTGGG TTGGATGATGGGTCCATGGTTGGTTTATGCTTCTTTGTTGAATGGTGCTTCTATTGCTTTGTATAATGGTTCTCC ATTGGTTTCTGGTTTTGCTAAATTTGTTCAAGATGCTAAAGTTACTATGTTGGGTGTTGTTCCATCTATTGTTAGA TCTTGGAAATCTACTAATTGTGTTTCTGGTTATGATTGGTCTACTATTAGATGTTTTTCTTCTTCTGGTGAAGCTT CTAATGTTGATGAATATTTGTGGTTGATGGGTAGAGCTAATTATAAACCAGTTATTGAAATGTGTGGTGGTACTG AAATTGGTGGTGCTTTTTCTGCTGGTTCTTTTTTGCAAGCTCAATCTTTGTCTTCTTTTTCTTCTCAATGTATGGG TTGTACTTTGTATATTTTGGATAAAAATGGTTATCCAATGCCAAAAAATAAACCAGGTATTGGTGAATTGGCTTTG GGTCCAGTTATGTTTGGTGCTTCTAAAACTTTGTTGAATGGTAATCATCATGATGTTTATTTTAAAGGTATGCCAA CTTTGAATGGTGAAGTTTTGAGAAGACATGGTGATATTTTTGAATTGACTTCTAATGGTTATTATCATGCTCATGG TAGAGCTGATGATACTATGAATATTGGTGGTATTAAAATTTCTTCTATTGAAATTGAAAGAGTTTGTAATGAAGTT GATGATAGAGTTTTTGAAACTACTGCTATTGGTGTTCCACCATTGGGTGGTGGTCCAGAACAATTGGTTATTTTT TTTGTTTTGAAAGATTCTAATGATACTACTATTGATTTGAATCAATTGAGATTGTCTTTTAATTTGGGTTTGCAAAA AAAATTGAATCCATTGTTTAAAGTTACTAGAGTTGTTCCATTGTCTTCTTTGCCAAGAACTGCTACTAATAAAATT ATGAGAAGAGTTTTGAGACAACAATTTTCTCATTTTGAA

Figure 12B

ATP Citrate Lyase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID1

ATGTCTGCTAAAGCTATTTCTGAACAAACTGGTAAAGAATTGTTGTATAAATTTATTTGTACTACTTCTGCTATTC AAAATAGATTTAAATATGCTAGAGTTACTCCAGATACTGATTGGGCTAGATTGTTGCAAGATCATCCATGGTTGT TGTCTCAAAATTTGGTTGTTAAACCAGATCAATTGATTAAAAGAAGAGGTAAATTGGGTTTGGTTGGTGTTAATTT GACTTTGGATGGTGTTAAATCTTGGTTGAAACCAAGATTGGGTCAAGAAGCTACTGTTGGTAAAGCTACTGGTT TTTTGAAAAATTTTTTGATTGAACCATTTGTTCCACATTCTCAAGCTGAAGAATTTTATGTTTGTATTTATGCTACT AGAGAAGGTGATTATGTTTTGTTTCATCATGAAGGTGGTGTTGATGTTGGTGATGTTGATGCTAAAGCTCAAAAA TTGTTGGTTGGTGTTGATGAAAAATTGAATCCAGAAGATATTAAAAAACATTTGTTGGTTCATGCTCCAGAAGAT AAAAAAGAAATTTTGGCTTCTTTTATTTCTGGTTTGTTTAATTTTTATGAAGATTTGTATTTTACTTATTTGGAAATT AATCCATTGGTTGTTACTAAAGATGGTGTTTATGTTTTGGATTTGGCTGCTAAAGTTGATGCTACTGCTGATTATA TTTGTAAAGTTAAATGGGGTGATATTGAATTTCCACCACCATTTGGTAGAGAAGCTTATCCAGAAGAAGCTTATA TTGCTGATTTGGATGCTAAATCTGGTGCTTCTTTGAAATTGACTTTGTTGAATCCAAAAGGTAGAATTTGGACTAT GGTTGCTGGTGGTGGTGCTTCTGTTGTTTATTCTGATACTATTTGTGATTTGGGTGGTGTTAATGAATTGGCTAA TTATGGTGAATATTCTGGTGCTCCATCTGAACAACAAACTTATGATTATGCTAAAACTATTTTGTCTTTGATGACT AGAGAAAAACATCCAGATGGTAAAATTTTGATTATTGGTGGTTCTATTGCTAATTTTACTAATGTTGCTGCTACTT TTAAAGGTATTGTTAGAGCTATTAGAGATTATCAAGGTCCATTGAAAGAACATGAAGTTACTATTTTTGTTAGAAG AGGTGGTCCAAATTATCAAGAAGGTTTGAGAGTTATGGGTGAAGTTGGTAAAACTACTGGTATTCCAATTCATGT TTTTGGTACTGAAACTCATATGACTGCTATTGTTGGTATGGCTTTGGGTCATAGACCAATTCCAAATCAACCACC AACTGCTGCTCATACTGCTAATTTTTTGTTGAATGCTTCTGGTTCTACTTCTACTCCAGCTCCATCTAGAACTGCT TCTTTTTCTGAATCTAGAGCTGATGAAGTTGCTCCAGCTAAAAAAGCTAAACCAGCTATGCCACAAGATTCTGTT CCATCTCCAAGATCTTTGCAAGGTAAATCTACTACTTTGTTTTCTAGACATACTAAAGCTATTGTTTGGGGTATGC AAACTAGAGCTGTTCAAGGTATGTTGGATTTTGATTATGTTTGTTCTAGAGATGAACCATCTGTTGCTGCTATGG TTTATCCATTTACTGGTGATCATAAACAAAAATTTTATTGGGGTCATAAAGAAATTTTGATTCCAGTTTTTAAAAAT ATGGCTGATGCTATGAGAAAACATCCAGAAGTTGATGTTTTGATTAATTTTGCTTCTTTGAGATCTGCTTATGATT CTACTATGGAAACTATGAATTATGCTCAAATTAGAACTATTGCTATTATTGCTGAAGGTATTCCAGAAGCTTTGAC TAGAAAATTGATTAAAAAAGCTGATCAAAAAGGTGTTACTATTATTGGTCCAGCTACTGTTGGTGGTATTAAACC AGGTTGTTTTAAAATTGGTAATACTGGTGGTATGTTGGATAATATTTTGGCTTCTAAATTGTATAGACCAGGTTCT

Figure 12B (continued)

GTTGCTTATGTTTCTAGATCTGGTGGTATGTCTAATGAATTGAATAATATTATTTCTAGAACTACTGATGGTGTTT ATGAAGGTGTTGCTATTGGTGGTGATAGATATCCAGGTTCTACTTTTATGGATCATGTTTTGAGATATCAAGATA CTCCAGGTGTTAAAATGATTGTTGTTTTGGGTGAAATTGGTGGTACTGAAGAATATAAAATTTGTAGAGGTATTA AAGAAGGTAGATTGACTAAACCAATTGTTTGTTGGTGTATTGGTACTTGTGCTACTATGTTTTCTTCTGAAGTTCA ATTTGGTCATGCTGGTGCTTGTGCTAATCAAGCTTCTGAAACTGCTGTTGCTAAAAATCAAGCTTTGAAAGAAGC TGGTGTTTTTGTTCCAAGATCTTTTGATGAATTGGGTGAAATTATTCAATCTGTTTATGAAGATTTGGTTGCTAAT GGTGTTATTGTTCCAGCTCAAGAAGTTCCACCACCAACTGTTCCAATGGATTATTCTTGGGCTAGAGAATTGGG TTTGATTAGAAAACCAGCTTCTTTTATGACTTCTATTTGTGATGAAAGAGGTCAAGAATTGATTTATGCTGGTATG CCAATTACTGAAGTTTTTAAAGAAGAAATGGGTATTGGTGGTGTTTTGGGTTTGTTGTGGTTTCAAAAAAGATTG CCAAAATATTCTTGTCAATTTATTGAAATGTGTTTGATGGTTACTGCTGATCATGGTCCAGCTGTTTCTGGTGCT CATAATACTATTATTTGTGCTAGAGCTGGTAAAGATTTGGTTTCTTCTTTGACTTCTGGTTTGTTGACTATTGGTG ATAGATTTGGTGGTGCTTTGGATGCTGCTGCTAAAATGTTTTCTAAAGCTTTTGATTCTGGTATTATTCCAATGGA ATTTGTTAATAAAATGAAAAAAGAAGGTAAATTGATTATGGGTATTGGTCATAGAGTTAAATCTATTAATAATCCA GATATGAGAGTTCAAATTTTGAAAGATTATGTTAGACAACATTTTCCAGCTACTCCATTGTTGGATTATGCTTTGG AAGTTGAAAAAATTACTACTTCTAAAAAACCAAATTTGATTTTGAATGTTGATGGTTTGATTGGTGTTGCTTTTGTT GATATGTTGAGAAATTGTGGTTCTTTTACTAGAGAAGAAGCTGATGAATATATTGATATTGGTGCTTTGAATGGT ATTTTTGTTTTGGGTAGATCTATGGGTTTTATTGGTCATTATTTGGATCAAAAAAGATTGAAACAAGGTTTGTATA GACATCCATGGGATGATATTTCTTATGTTTTGCCAGAACATATGTCTATGAAATTGTCTGGTGGTGGTGGTTCTG GTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGT GATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATT TGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCC AAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGAT TATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATA AACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTG GTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAA TTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCT ATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGT GGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTG TATAAAAAAGCTGGTTCTGGTTCTAATTTGGTTGCTCAATTGGAAAATGAAGTTGCTTCTTTGGAAAATGAAAAT GAAACTTTGAAAAAAAAAAATTTGCATAAAAAAGATTTGATTGCTTATTTGGAAAAAGAAATTGCTAATTTGAGAA AAAAAATTGAAGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAA GAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCAT CATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTGCTAGAAATGCTTATTTGAGAA AAAAAATTGCTAGATTGAAAAAAGATAATTTGCAATTGGAAAGAGATGAACAAAATTTGGAAAAAATTATTGCTAA TTTGAGAGATGAAATTGCTAGATTGGAAAATGAAGTTGCTTCTCATGAACAA

Acetyl-CoA Acetyltransferase (atoB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID2

ATGAAAAATTGTGTTATTGTTTCTGCTGTTAGAACTGCTATTGGTTCTTTTAATGGTTCTTTGGCTTCTACTTCTG CTATTGATTTGGGTGCTACTGTTATTAAAGCTGCTATTGAAAGAGCTAAAATTGATTCTCAACATGTTGATGAAGT TATTATGGGTAATGTTTTGCAAGCTGGTTTGGGTCAAAATCCAGCTAGACAAGCTTTGTTGAAATCTGGTTTGGC TGAAACTGTTTGTGGTTTTACTGTTAATAAAGTTTGTGGTTCTGGTTTGAAATCTGTTGCTTTGGCTGCTCAAGCT ATTCAAGCTGGTCAAGCTCAATCTATTGTTGCTGGTGGTATGGAAAATATGTCTTTGGCTCCATATTTGTTGGAT GCTAAAGCTAGATCTGGTTATAGATTGGGTGATGGTCAAGTTTATGATGTTATTTTGAGAGATGGTTTGATGTGT GCTACTCATGGTTATCATATGGGTATTACTGCTGAAAATGTTGCTAAAGAATATGGTATTACTAGAGAAATGCAA GATGAATTGGCTTTGCATTCTCAAAGAAAAGCTGCTGCTGCTATTGAATCTGGTGCTTTTACTGCTGAAATTGTT CCAGTTAATGTTGTTACTAGAAAAAAAACTTTTGTTTTTTCTCAAGATGAATTTCCAAAAGCTAATTCTACTGCTG AAGCTTTGGGTGCTTTGAGACCAGCTTTTGATAAAGCTGGTACTGTTACTGCTGGTAATGCTTCTGGTATTAATG ATGGTGCTGCTGCTTTGGTTATTATGGAAGAATCTGCTGCTTTGGCTGCTGGTTTGACTCCATTGGCTAGAATTA AATCTTATGCTTCTGGTGGTGTTCCACCAGCTTTGATGGGTATGGGTCCAGTTCCAGCTACTCAAAAAGCTTTG CAATTGGCTGGTTTGCAATTGGCTGATATTGATTTGATTGAAGCTAATGAAGCTTTTGCTGCTCAATTTTTGGCT

Figure 12B (continued)

GTTGGTAAAAATTTGGGTTTTGATTCTGAAAAAGTTAATGTTAATGGTGGTGCTATTGCTTTGGGTCATCCAATT GGTGCTTCTGGTGCTAGAATTTTGGTTACTTTGTTGCATGCTATGCAAGCTAGAGATAAAACTTTGGGTTTGGCT ACTTTGTGTATTGGTGGTGGTCAAGGTATTGCTATGGTTATTGAAAGATTGAATAAATTGTCTGGTGGTGGTGGT TCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACA AGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTA TAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTG GATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTG AAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTT ATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTC TGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCT TTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAA AAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGT TCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACT TCTTTGTATAAAAAAGCTGGTTCTGGTTCTAATGAAGTTACTACTTTGGAAAATGATGCTGCTTTTATTGAAAATG AAAATGCTTATTTGGAAAAAGAAATTGCTAGATTGAGAAAAGAAAAAGCTGCTTTGAGAAATAGATTGGCTCATA AAAAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCT GCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCAT CATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTGCTGAATTGAAAAATAGAGTTG CTGTTAAATTGAATAGAAATGAACAATTGAAAAATAAAGTTGAAGAATTGAAAAATAGAAATGCTTATTTGAAAAA TGAATTGGCTACTTTGGAAAATGAAGTTGCTAGATTGGAAAATGATGTTGCTGAA

3-Hvdroxvbutvrvl-CoA Dehydrogenase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID3

ATGAAAAAAGTTTGTGTTATTGGTGCTGGTACTATGGGTTCTGGTATTGCTCAAGCTTTTGCTGCTAAAGGTTTT GAAGTTGTTTTGAGAGATATTAAAGATGAATTTGTTGATAGAGGTTTGGATTTTATTAATAAAAATTTGTCTAAATT GGTTAAAAAAGGTAAAATTGAAGAAGCTACTAAAGTTGAAATTTTGACTAGAATTTCTGGTACTGTTGATTTGAAT ATGGCTGCTGATTGTGATTTGGTTATTGAAGCTGCTGTTGAAAGAATGGATATTAAAAAACAAATTTTTGCTGATT TGGATAATATTTGTAAACCAGAAACTATTTTGGCTTCTAATACTTCTTCTTTGTCTATTACTGAAGTTGCTTCTGCT ACTAAAAGACCAGATAAAGTTATTGGTATGCATTTTTTTAATCCAGCTCCAGTTATGAAATTGGTTGAAGTTATTA GAGGTATTGCTACTTCTCAAGAAACTTTTGATGCTGTTAAAGAAACTTCTATTGCTATTGGTAAAGATCCAGTTG AAGTTGCTGAAGCTCCAGGTTTTGTTGTTAATAGAATTTTGATTCCAATGATTAATGAAGCTGTTGGTATTTTGGC TGAAGGTATTGCTTCTGTTGAAGATATTGATAAAGCTATGAAATTGGGTGCTAATCATCCAATGGGTCCATTGGA ATTGGGTGATTTTATTGGTTTGGATATTTGTTTGGCTATTATGGATGTTTTGTATTCTGAAACTGGTGATTCTAAA TATAGACCACATACTTTGTTGAAAAAATATGTTAGAGCTGGTTGGTTGGGTAGAAAATCTGGTAAAGGTTTTTAT GATTATTCTAAAAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGC TTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTG GAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAA GCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTT ATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCA AGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAA AAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATT ATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAG GTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAA TAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGAAAATT TGTATTTTCAAGGTGAAAATTTGTATTTTCAAGGTGATTCTTCTGAATCTTGTTGGAATTGTGGTAGAAAAGCTTC TGAAACTTGTTCTGGTTGTAATACTGCTAGATATTGTGGTTCTTTTTGTCAACATAAAGATTGGGAAAAACATCAT CATATTTGTGGTCAAACTTTGCAAGCTCAACAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCT GCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTG GTTCTATGGCTGTTTCTGAATCTCAATTGAAAAAAATGGTTTCTAAATATAAATATAGAGATTTGACTGTTAGAGA AACTGTTAATGTTATTACTTTGTATAAAGATTTGAAACCAGTTTTGGATTCTTATGTTTTTAATGATGGTTCTTCTA GAGAATTGATGAATTTGACTGGTACTATTCCAGTTCCATATAGAGGTAATACTTATAATATTCCAATTTGTTTGTG

Figure 12B (continued)

GTTGTTGGATACTTATCCATATAATCCACCAATTTGTTTTGTTAAACCAACTTCTTCTATGACTATTAAAACTGGTA AACATGTTGATGCTAATGGTAAAATTTATTTGCCATATTTGCATGAATGGAAACATCCACAATCTGATTTGTTGGG TTTGATTCAAGTTATGATTGTTGTTTTTGGTGATGAACCACCAGTTTTTTCTAGACCA

Enoyl-CoA Hydratase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID4

ATGGAATTGAATAATGTTATTTTGGAAAAAGAAGGTAAAGTTGCTGTTGTTACTATTAATAGACCAAAAGCTTTGA ATGCTTTGAATTCTGATACTTTGAAAGAAATGGATTATGTTATTGGTGAAATTGAAAATGATTCTGAAGTTTTGGC TGTTATTTTGACTGGTGCTGGTGAAAAATCTTTTGTTGCTGGTGCTGATATTTCTGAAATGAAAGAAATGAATACT ATTGAAGGTAGAAAATTTGGTATTTTGGGTAATAAAGTTTTTAGAAGATTGGAATTGTTGGAAAAACCAGTTATTG CTGCTGTTAATGGTTTTGCTTTGGGTGGTGGTTGTGAAATTGCTATGTCTTGTGATATTAGAATTGCTTCTTCTAA TGCTAGATTTGGTCAACCAGAAGTTGGTTTGGGTATTACTCCAGGTTTTGGTGGTACTCAAAGATTGTCTAGATT GGTTGGTATGGGTATGGCTAAACAATTGATTTTTACTGCTCAAAATATTAAAGCTGATGAAGCTTTGAGAATTGG TTTGGTTAATAAAGTTGTTGAACCATCTGAATTGATGAATACTGCTAAAGAAATTGCTAATAAAATTGTTTCTAAT GCTCCAGTTGCTGTTAAATTGTCTAAACAAGCTATTAATAGAGGTATGCAATGTGATATTGATACTGCTTTGGCT TTTGAATCTGAAGCTTTTGGTGAATGTTTTTCTACTGAAGATCAAAAAGATGCTATGACTGCTTTTATTGAAAAAA GAAAAATTGAAGGTTTTAAAAATAGAAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGT GGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATT ATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAG GTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAA TTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGAT CCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCT ATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATT ATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGC TTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTG GAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTG GTGCTTCTGGTCCATTGGGTTCTCCATTGACTGCTTCTATGTTGGCTTCTGCTCCACCACAAGAACAAAAACAAA TGTTGGGTGAAAGATTGTTTCCATTGATTCAAGCTATGCATCCAACTTTGGCTGGTAAAATTACTGGTATGTTGT TGGAAATTGATAATTCTGAATTGTTGCATATGTTGGAATCTCCAGAATCTTTGAGATCTAAAGTTGATGAAGCTG TTGCTGTTTTGCAAGCTCATCAAGCTAAAGAAGCTGCTCAAAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCT GGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTG GTTCTGGTGAATTTGGTTCTAATACTAATATGTCTGTTCCAACTGATGGTGCTGTTACTACTTCTCAAATTCCAGC TTCTGAACAAGAAACTTTGGTTAGACCAAAACCATTGTTGTTGAAATTGTTGAAATCTGTTGGTGCTCAAAAAGA TACTTATACTATGAAAGAAGTTTTGTTTTATTTGGGTCAATATATTATGACTAAAAGATTGTATGATGAAAAACAAC AACATATTGTTTATTGTTCTAATGATTTGTTGGGTGATTTGTTTGGTGTTCCATCTTTTTCTGTTAAAGAACATAGA AAAATTTATACTATGATTTATAGAAATTTGGTTGTT

Trans-Enoyl-CoA Reductase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID5

ATGATTGTTAAACCAATGGTTAGAAATAATATTTGTTTGAATGCTCATCCACAAGGTTGTAAAAAAGGTGTTGAA GATCAAATTGAATATACTAAAAAAAGAATTACTGCTGAAGTTAAAGCTGGTGCTAAAGCTCCAAAAAATGTTTTG GTTTTGGGTTGTTCTAATGGTTATGGTTTGGCTTCTAGAATTACTGCTGCTTTTGGTTATGGTGCTGCTACTATT GGTGTTTCTTTTGAAAAAGCTGGTTCTGAAACTAAATATGGTACTCCAGGTTGGTATAATAATTTGGCTTTTGAT GAAGCTGCTAAAAGAGAAGGTTTGTATTCTGTTACTATTGATGGTGATGCTTTTTCTGATGAAATTAAAGCTCAA GTTATTGAAGAAGCTAAAAAAAAAGGTATTAAATTTGATTTGATTGTTTATTCTTTGGCTTCTCCAGTTAGAACTG ATCCAGATACTGGTATTATGCATAAATCTGTTTTGAAACCATTTGGTAAAACTTTTACTGGTAAAACTGTTGATCC ATTTACTGGTGAATTGAAAGAAATTTCTGCTGAACCAGCTAATGATGAAGAAGCTGCTGCTACTGTTAAAGTTAT GGGTGGTGAAGATTGGGAAAGATGGATTAAACAATTGTCTAAAGAAGGTTTGTTGGAAGAAGGTTGTATTACTT TGGCTTATTCTTATATTGGTCCAGAAGCTACTCAAGCTTTGTATAGAAAAGGTACTATTGGTAAAGCTAAAGAAC ATTTGGAAGCTACTGCTCATAGATTGAATAAAGAAAATCCATCTATTAGAGCTTTTGTTTCTGTTAATAAAGGTTT GGTTACTAGAGCTTCTGCTGTTATTCCAGTTATTCCATTGTATTTGGCTTCTTTGTTTAAAGTTATGAAAGAAAAA

Figure 12B (continued)

GGTAATCATGAAGGTTGTATTGAACAAATTACTAGATTGTATGCTGAAAGATTGTATAGAAAAGATGGTACTATT CCAGTTGATGAAGAAAATAGAATTAGAATTGATGATTGGGAATTGGAAGAAGATGTTCAAAAAGCTGTTTCTGCT TTGATGGAAAAAGTTACTGGTGAAAATGCTGAATCTTTGACTGATTTGGCTGGTTATAGACATGATTTTTTGGCT TCTAATGGTTTTGATGTTGAAGGTATTAATTATGAAGCTGAAGTTGAAAGATTTGATAGAATTAAATTGTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTAT TATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAG CTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTT GGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAA AGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGG TAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAAT AATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATC AAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTG ATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTG GTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCTTATTATCATCATCATCATCATCATTTG GAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTAATTTGTTGGCTACTTTGAGATCTACTGCTGCTGTTT TGGAAAATGAAAATCATGTTTTGGAAAAAGAAAAAGAAAAATTGAGAAAAGAAAAAGAACAATTGTTGAATAAATT GGAAGCTTATAAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAG AAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATC ATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTAAAAGAATTGCTTATTTGAGAAA AAAAATTGCTGCTTTGAAAAAAGATAATGCTAATTTGGAAAAAGATATTGCTAATTTGGAAAATGAAATTGAAAGA TTGATTAAAGAAATTAAAACTTTGGAAAATGAAGTTGCTTCTCATGAACAA

Beta-Ketothiolase (bktB) - Enzyme Linker - 0TPR6 Spacer - ID Linker - ID6

ATGACTAGAGAAGTTGTTGTTGTTTCTGGTGTTAGAACTGCTATTGGTACTTTTGGTGGTTCTTTGAAAGATGTT GCTCCAGCTGAATTGGGTGCTTTGGTTGTTAGAGAAGCTTTGGCTAGAGCTCAAGTTTCTGGTGATGATGTTGG TCATGTTGTTTTTGGTAATGTTATTCAAACTGAACCAAGAGATATGTATTTGGGTAGAGTTGCTGCTGTTAATGG TGGTGTTACTATTAATGCTCCAGCTTTGACTGTTAATAGATTGTGTGGTTCTGGTTTGCAAGCTATTGTTTCTGCT GCTCAAACTATTTTGTTGGGTGATACTGATGTTGCTATTGGTGGTGGTGCTGAATCTATGTCTAGAGCTCCATAT TTGGCTCCAGCTGCTAGATGGGGTGCTAGAATGGGTGATGCTGGTTTGGTTGATATGATGTTGGGTGCTTTGC ATGATCCATTTCATAGAATTCATATGGGTGTTACTGCTGAAAATGTTGCTAAAGAATATGATATTTCTAGAGCTCA ACAAGATGAAGCTGCTTTGGAATCTCATAGAAGAGCTTCTGCTGCTATTAAAGCTGGTTATTTTAAAGATCAAAT TGTTCCAGTTGTTTCTAAAGGTAGAAAAGGTGATGTTACTTTTGATACTGATGAACATGTTAGACATGATGCTAC TATTGATGATATGACTAAATTGAGACCAGTTTTTGTTAAAGAAAATGGTACTGTTACTGCTGGTAATGCTTCTGGT TTGAATGATGCTGCTGCTGCTGTTGTTATGATGGAAAGAGCTGAAGCTGAAAGAAGAGGTTTGAAACCATTGGC TAGATTGGTTTCTTATGGTCATGCTGGTGTTGATCCAAAAGCTATGGGTATTGGTCCAGTTCCAGCTACTAAAAT TGCTTTGGAAAGAGCTGGTTTGCAAGTTTCTGATTTGGATGTTATTGAAGCTAATGAAGCTTTTGCTGCTCAAGC TTGTGCTGTTACTAAAGCTTTGGGTTTGGATCCAGCTAAAGTTAATCCAAATGGTTCTGGTATTTCTTTGGGTCA TCCAATTGGTGCTACTGGTGCTTTGATTACTGTTAAAGCTTTGCATGAATTGAATAGAGTTCAAGGTAGATATGC TTTGGTTACTATGTGTATTGGTGGTGGTCAAGGTATTGCTGCTATTTTTGAAAGAATTAAATTGTCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATA AACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTT GGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGA ATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCT ATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAAT GCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATG CTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAA AGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTA TCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGATGTTATGTGGGAATATAAATGGGAAAATACTG GTGATGCTGAATTGTATGGTCCATTTACTTCTGCTCAAATGCAAACTTGGGTTTCTGAAGGTTATTTTCCAGATG GTGTTTATTGTAGAAAATTGGATCCACCAGGTGGTCAATTTTATAATTCTAAAAGAATTGATTTTGATTTGTATAC TGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTA

Figure 12B (continued)

AAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGAATCTGATTCTGTTGAATTTAATAATG CTATTTCTTATGTTAATAAAATTAAAACTAGATTTTTGGATCATCCAGAAATTTATAGATCTTTTTTGGAAATTTTG CATACTTATCAAAAAGAACAATTGCATACTAAAGGTAGACCATTTAGAGGTATGTCTGAAGAAGAAGTTTTTACT GAAGTTGCTAATTTGTTTAGAGGTCAAGAAGATTTGTTGTCTGAATTTGGTCAATTTTTGCCAGAAGCTAAAAGA

HMG CoA Synthase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID7

ATGAAACTCTCAACTAAACTTTGTTGGTGTGGTATTAAAGGAAGACTTAGGCCGCAAAAGCAACAACAATTACAC AATACAAACTTGCAAATGACTGAACTAAAAAAACAAAAGACCGCTGAACAAAAAACCAGACCTCAAAATGTCGGT ATTAAAGGTATCCAAATTTACATCCCAACTCAATGTGTCAACCAATCTGAGCTAGAGAAATTTGATGGCGTTTCT CAAGGTAAATACACAATTGGTCTGGGCCAAACCAACATGTCTTTTGTCAATGACAGAGAAGATATCTACTCGAT GTCCCTAACTGTTTTGTCTAAGTTGATCAAGAGTTACAACATCGACACCAACAAAATTGGTAGATTAGAAGTCGG TACTGAAACTCTGATTGACAAGTCCAAGTCTGTCAAGTCTGTCTTGATGCAATTGTTTGGTGAAAACACTGACGT CGAAGGTATTGACACGCTTAATGCCTGTTACGGTGGTACCAACGCGTTGTTCAACTCTTTGAACTGGATTGAAT CTAACGCATGGGATGGTAGAGACGCCATTGTAGTTTGCGGTGATATTGCCATCTACGATAAGGGTGCCGCAAG ACCAACCGGTGGTGCCGGTACTGTTGCTATGTGGATCGGTCCTGATGCTCCAATTGTATTTGACTCTGTAAGAG CTTCTTACATGGAACACGCCTACGATTTTTACAAGCCAGATTTCACCAGCGAATATCCTTACGTCGATGGTCATT TTTCATTAACTTGTTACGTCAAGGCTCTTGATCAAGTTTACAAGAGTTATTCCAAGAAGGCTATTTCTAAAGGGTT GGTTAGCGATCCCGCTGGTTCGGATGCTTTGAACGTTTTGAAATATTTCGACTACAACGTTTTCCATGTTCCAAC CTGTAAATTGGTCACAAAATCATACGGTAGATTACTATATAACGATTTCAGAGCCAATCCTCAATTGTTCCCAGA AGTTGACGCCGAATTAGCTACTCGCGATTATGACGAATCTTTAACCGATAAGAACATTGAAAAAACTTTTGTTAA TGTTGCTAAGCCATTCCACAAAGAGAGAGTTGCCCAATCTTTGATTGTTCCAACAAACACAGGTAACATGTACAC CGCATCTGTTTATGCCGCCTTTGCATCTCTATTAAACTATGTTGGATCTGACGACTTACAAGGCAAGCGTGTTG GTTTATTTTCTTACGGTTCCGGTTTAGCTGCATCTCTATATTCTTGCAAAATTGTTGGTGACGTCCAACATATTAT CAAGGAATTAGATATTACTAACAAATTAGCCAAGAGAATCACCGAAACTCCAAAGGATTACGAAGCTGCCATCG AATTGAGAGAAAATGCCCATTTGAAGAAGAACTTCAAACCTCAAGGTTCCATTGAGCATTTGCAAAGTGGTGTTT ACTACTTGACCAACATCGATGACAAATTTAGAAGATCTTACGATGTTAAAAAATAAAAATTGTCTGGTGGTGGTG GTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAA CAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGG TATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAAT TGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTAT TGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGC TTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCT TCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAG CTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATC AAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTG GTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTTGGGTCCATTGCCACCAGGTTGGGAAGTTAGATC TACTGTTTCTGGTAGAATTTATTTTGTTGATCATAATAATAGAACTACTCAATTTACTGATCCAAGATTGCATGGT TCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGC TGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGGTGCTATGGGTCCATTGCCACCAGGTTGG GAAAAAAGAACTGATTCTAATGGTAGAGTTTATTTTGTTAATCATAATACTAGAATTACTCAATGGGAAGATCCAA GATCT

Truncated HMG-CoA Reductase - Enzyme Linker - cTPR6 Spacer - ID Linker - IDS

ATGGTTGCGGTACGTAGGAAGGCTCTTTCAATTTTGGCAGAAGCTCCTGTATTAGCATCTGATCGTTTACCATAT AAAAATTATGACTACGACCGCGTATTTGGCGCTTGTTGTGAAAATGTTATAGGTTACATGCCTTTGCCCGTTGGT GTTATAGGCCCCTTGGTTATCGATGGTACATCTTATCATATACCAATGGCAACTACAGAGGGTTGTTTGGTAGCT TCTGCCATGCGTGGCTGTAAGGCAATCAATGCTGGCGGTGGTGCAACAACTGTTTTAACTAAGGATGGTATGA CAAGAGGCCCAGTAGTCCGTTTCCCAACTTTGAAAAGATCTGGTGCCTGTAAGATATGGTTAGACTCAGAAGAG GGACAAAACGCAATTAAAAAAGCTTTTAACTCTACATCAAGATTTGCACGTCTGCAACATATTCAAACTTGTCTA

Figure 12B (continued)

GCAGGAGATTTACTCTTCATGAGATTTAGAACAACTACTGGTGACGCAATGGGTATGAATATGATTTCTAAAGGT GTCGAATACTCATTAAAGCAAATGGTAGAAGAGTATGGCTGGGAAGATATGGAGGTTGTCTCCGTTTCTGGTAA CTACTGTACCGACAAAAAACCAGCTGCCATCAACTGGATCGAAGGTCGTGGTAAGAGTGTCGTCGCAGAAGCT ACTATTCCTGGTGATGTTGTCAGAAAAGTGTTAAAAAGTGATGTTTCCGCATTGGTTGAGTTGAACATTGCTAAG AATTTGGTTGGATCTGCAATGGCTGGGTCTGTTGGTGGATTTAACGCACATGCAGCTAATTTAGTGACAGCTGT TTTCTTGGCATTAGGACAAGATCCTGCACAAAATGTTGAAAGTTCCAACTGTATAACATTGATGAAAGAAGTGGA CGGTGATTTGAGAATTTCCGTATCCATGCCATCCATCGAAGTAGGTACCATCGGTGGTGGTACTGTTCTAGAAC CACAAGGTGCCATGTTGGACTTATTAGGTGTAAGAGGCCCGCATGCTACCGCTCCTGGTACCAACGCACGTCA ATTAGCAAGAATAGTTGCCTGTGCCGTCTTGGCAGGTGAATTATCCTTATGTGCTGCCCTAGCAGCCGGCCATT TGGTTCAAAGTCATATGACCCACAACAGGAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGG TGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGA ATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAA CAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGG TATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAAT TGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAA AAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGC TTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCT AAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAA GCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTG GTGGTGGTGCTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTC TGAATTTTTTAGAAGAGAAAGAAATAAAATGGCTGCTGCTAAATGTAGAAATAGAAGAAGAGAATTGACTGATAC TTTGCAAGCTGAAACTGATCAATTGGAAGATGAAAAATCTGCTTTGCAAACTGAAATTGCTAATTTGTTGAAAGA AAAAGAAAAATTGGAATTTATTTTGGCTGCTCATAGACCAGCTTGTAAAATTCCAGATGATTTGGGTTTTCCAGA AGAAATGTCTTTGGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTA AAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATC ATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTGAATCTTTGAA ACAAAAAATTGAAGAATTGAAACAAAGAAAAGCTCAATTGAAAAATGATATTGCTAATTTGGAAAAAGAAATTGCT TATGCTGAAACT

Mevalonate Kinase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID9

ATGTCATTACCGTTCTTAACTTCTGCACCGGGAAAGGTTATTATTTTTGGTGAACACTCTGCTGTGTACAACAAG CCTGCCGTCGCTGCTAGTGTGTCTGCGTTGAGAACCTACCTGCTAATAAGCGAGTCATCTGCACCAGATACTAT TGAATTGGACTTCCCGGACATTAGCTTTAATCATAAGTGGTCCATCAATGATTTCAATGCCATCACCGAGGATCA AGTAAACTCCCAAAAATTGGCCAAGGCTCAACAAGCCACCGATGGCTTGTCTCAGGAACTCGTTAGTCTTTTGG ATCCGTTGTTAGCTCAACTATCCGAATCCTTCCACTACCATGCAGCGTTTTGTTTCCTGTATATGTTTGTTTGCCT ATGCCCCCATGCCAAGAATATTAAGTTTTCTTTAAAGTCTACTTTACCCATCGGTGCTGGGTTGGGCTCAAGCG CCTCTATTTCTGTATCACTGGCCTTAGCTATGGCCTACTTGGGGGGGTTAATAGGATCTAATGACTTGGAAAAG CTGTCAGAAAACGATAAGCATATAGTGAATCAATGGGCCTTCATAGGTGAAAAGTGTATTCACGGTACCCCTTC AGGAATAGATAACGCTGTGGCCACTTATGGTAATGCCCTGCTATTTGAAAAAGACTCACATAATGGAACAATAAA CACAAACAATTTTAAGTTCTTAGATGATTTCCCAGCCATTCCAATGATCCTAACCTATACTAGAATTCCAAGGTCT ACAAAAGATCTTGTTGCTCGCGTTCGTGTGTTGGTCACCGAGAAATTTCCTGAAGTTATGAAGCCAATTCTAGAT GCCATGGGTGAATGTGCCCTACAAGGCTTAGAGATCATGACTAAGTTAAGTAAATGTAAAGGCACCGATGACGA GGCTGTAGAAACTAATAATGAACTGTATGAACAACTATTGGAATTGATAAGAATAAATCATGGACTGCTTGTCTC AATCGGTGTTTCTCATCCTGGATTAGAACTTATTAAAAATCTGAGCGATGATTTGAGAATTGGCTCCACAAAACT TACCGGTGCTGGTGGCGGCGGTTGCTCTTTGACTTTGTTACGAAGAGACATTACTCAAGAGCAAATTGACAGCT TCAAAAAGAAATTGCAAGATGATTTTAGTTACGAGACATTTGAAACAGACTTGGGTGGGACTGGCTGCTGTTTG TTAAGCGCAAAAAATTTGAATAAAGATCTTAAAATCAAATCCCTAGTATTCCAATTATTTGAAAATAAAACTACCA CAAAGCAACAAATTGACGATCTATTATTGCCAGGAAACACGAATTTACCATGGACTTCATAAAAATTGTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTAT TATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAG

Figure 12B (continued)

CTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTT GGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAA AGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGG TAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAAT AATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATC AAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTG ATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTG GTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTATGGAACCAGCTATGGAACCAGAAACTTT GGAAGCTAGAATTAATAGAGCTACTAATCCATTGAATAAAGAATTGGATTGGGCTTCTATTAATGGTTTTTGTGA ACAATTGAATGAAGATTTTGAAGGTCCACCATTGGCTACTAGATTGTTGGCTCATAAAATTCAATCTCCACAAGA ATGGGAAGCTATTCAAGCTTTGACTGTTTTGGAAACTTGTATGAAATCTTGTGGTAAAAGATTTCATGATGAAGT TGGTAAATTTAGATTTTTGAATGAATTGATTAAAGTTGTTTCTCCAAAATATTTGGGTTCTAGAACTTCTGAAAAA GTTAAAAATAAAATTTTGGAATTGTTGTATTCTTGGACTGTTGGTTTGCCAGAAGAAGTTAAAATTGCTGAAGCTT ATCAAATGTTGAAAAAACAAGGTATTGTTAAATCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTT CTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTT GGTTCTGGTGCTATGGGTTCTATGGCTGAAGCTGAAGGTGAATCTTTGGAATCTTGGTTGAATAAAGCTACTAA TCCATCTAATAGACAAGAAGATTGGGAATATATTATTGGTTTTTGTGATCAAATTAATAAAGAATTGGAAGGTCCA CAAATTGCTGTTAGATTGTTGGCTCATAAAATTCAATCTCCACAAGAATGGGAAGCTTTGCAAGCTTTGACTGTT TTGGAAGCTTGTATGAAAAATTGTGGTAGAAGATTTCATAATGAAGTTGGTAAATTTAGATTTTTGAATGAATTGA TTAAAGTTGTTTCTCCAAAATATTTGGGTGATAGAGTTTCTGAAAAAGTTAAAACTAAAGTTATTGAATTGTTGTA TTCTTGGACTATGGCTTTGCCAGAAGAAGCTAAAATTAAAGATGCTTATCATATGTTGAAAAGACAAGGTATTGT TCAATCTGATCCACCAATTCCAGTTGATAGAACTTTGATTCCATCTCCACCACCAAGACCAAAAAAT

Phosphomevalonate Kinase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID10

ATGTCAGAGTTGAGAGCCTTCAGTGCCCCAGGGAAAGCGTTACTAGCTGGTGGATATTTAGTTTTAGATACAAA ATATGAAGCATTTGTAGTCGGATTATCGGCAAGAATGCATGCTGTAGCCCATCCTTACGGTTCATTGCAAGGGT CTGATAAGTTTGAAGTGCGTGTGAAAAGTAAACAATTTAAAGATGGGGAGTGGCTGTACCATATAAGTCCTAAA AGTGGCTTCATTCCTGTTTCGATAGGCGGATCTAAGAACCCTTTCATTGAAAAAGTTATCGCTAACGTATTTAGC TACTTTAAACCTAACATGGACGACTACTGCAATAGAAACTTGTTCGTTATTGATATTTTCTCTGATGATGCCTACC ATTCTCAGGAGGATAGCGTTACCGAACATCGTGGCAACAGAAGATTGAGTTTTCATTCGCACAGAATTGAAGAA GTTCCCAAAACAGGGCTGGGCTCCTCGGCAGGTTTAGTCACAGTTTTAACTACAGCTTTGGCCTCCTTTTTTGT ATCGGACCTGGAAAATAATGTAGACAAATATAGAGAAGTTATTCATAATTTAGCACAAGTTGCTCATTGTCAAGC TCAGGGTAAAATTGGAAGCGGGTTTGATGTAGCGGCGGCAGCATATGGATCTATCAGATATAGAAGATTCCCA CCCGCATTAATCTCTAATTTGCCAGATATTGGAAGTGCTACTTACGGCAGTAAACTGGCGCATTTGGTTGATGA AGAAGACTGGAATATTACGATTAAAAGTAACCATTTACCTTCGGGATTAACTTTATGGATGGGCGATATTAAGAA TGGTTCAGAAACAGTAAAACTGGTCCAGAAGGTAAAAAATTGGTATGATTCGCATATGCCAGAAAGCTTGAAAA TATATACAGAACTCGATCATGCAAATTCTAGATTTATGGATGGACTATCTAAACTAGATCGCTTACACGAGACTC ATGACGATTACAGCGATCAGATATTTGAGTCTCTTGAGAGGAATGACTGTACCTGTCAAAAGTATCCTGAAATCA CAGAAGTTAGAGATGCAGTTGCCACAATTAGACGTTCCTTTAGAAAAATAACTAAAGAATCTGGTGCCGATATC GAACCTCCCGTACAAACTAGCTTATTGGATGATTGCCAGACCTTAAAAGGAGTTCTTACTTGCTTAATACCTGGT GCTGGTGGTTATGACGCCATTGCAGTGATTACTAAGCAAGATGTTGATCTTAGGGCTCAAACCGCTAATGACAA AAGATTTTCTAAGGTTCAATGGCTGGATGTAACTCAGGCTGACTGGGGTGTTAGGAAAGAAAAAGATCCGGAAA CTTATCTTGATAAATAAAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCT GAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAG CTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCA AAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAAT GCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATT TGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTA TCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGT GATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAA GAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATC CAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCT

Figure 12B (continued)

TATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTG AAGAATTGAAAAATAAAATTGCTGAATTGGAAAATAGAAATGCTGTTAAAAAAAATAGAGTTGCTCATTTGAAACA AGAAATTGCTTATTTGAAAGATGAATTGGCTGCTCATGAATTTGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGG TGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTT CTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGG TTCTGGTTCTTTTGAAAATGTTACTCATGAATTTATTTTGGCTACTTTGGAAAATGAAAATGCTAAATTGAGAAGA TTGGAAGCTAAATTGGAAAGAGAATTGGCTAGATTGAGAAATGAAGTTGCTTGGTTG

Diphosphomevalonate Decarboxylase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID11

ATGACCGTTTACACAGCATCCGTTACCGCACCCGTCAACATCGCAACCCTTAAGTATTGGGGGAAAAGGGACA CGAAGTTGAATCTGCCCACCAATTCGTCCATATCAGTGACTTTATCGCAAGATGACCTCAGAACGTTGACCTCT GCGGCTACTGCACCTGAGTTTGAACGCGACACTTTGTGGTTAAATGGAGAACCACACAGCATCGACAATGAAA GAACTCAAAATTGTCTGCGCGACCTACGCCAATTAAGAAAGGAAATGGAATCGAAGGACGCCTCATTGCCCAC ATTATCTCAATGGAAACTCCACATTGTCTCCGAAAATAACTTTCCTACAGCAGCTGGTTTAGCTTCCTCCGCTGC TGGCTTTGCTGCATTGGTCTCTGCAATTGCTAAGTTATACCAATTACCACAGTCAACTTCAGAAATATCTAGAAT AGCAAGAAAGGGGTCTGGTTCAGCTTGTAGATCGTTGTTTGGCGGATACGTGGCCTGGGAAATGGGAAAAGCT GAAGATGGTCATGATTCCATGGCAGTACAAATCGCAGACAGCTCTGACTGGCCTCAGATGAAAGCTTGTGTCCT AGTTGTCAGCGATATTAAAAAGGATGTGAGTTCCACTCAGGGTATGCAATTGACCGTGGCAACCTCCGAACTAT TTAAAGAAAGAATTGAACATGTCGTACCAAAGAGATTTGAAGTCATGCGTAAAGCCATTGTTGAAAAAGATTTCG CCACCTTTGCAAAGGAAACAATGATGGATTCCAACTCTTTCCATGCCACATGTTTGGACTCTTTCCCTCCAATAT TCTACATGAATGACACTTCCAAGCGTATCATCAGTTGGTGCCACACCATTAATCAGTTTTACGGAGAAACAATCG TTGCATACACGTTTGATGCAGGTCCAAATGCTGTGTTGTACTACTTAGCTGAAAATGAGTCGAAACTCTTTGCAT TTATCTATAAATTGTTTGGCTCTGTTCCTGGATGGGACAAGAAATTTACTACTGAGCAGCTTGAGGCTTTCAACC ATCAATTTGAATCATCTAACTTTACTGCACGTGAATTGGATCTTGAGTTGCAAAAGGATGTTGCCAGAGTGATTT TAACTCAAGTCGGTTCAGGCCCACAAGAAACAAACGAATCTTTGATTGACGCAAAGACTGGTCTACCAAAGGAA TAAAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATA ATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGA TCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAA TATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAAC AAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAG CTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTT GGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAA GCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCT TATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGAT CTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGCTATGGCTGATTT GGAACAAAAAGTTTTGGAAATGGAAGCTTCTACTTATGATGGTGTTTTTATTTGGAAAATTTCTGATTTTCCAAGA AAAAGACAAGAAGCTGTTGCTGGTAGAATTCCAGCTATTTTTTCTCCAGCTTTTTATACTTCTAGATATGGTTATA AAATGTGTTTGAGAATTTATTTGAATGGTGATGGTACTGGTAGAGGTACTCATTTGTCTTTGTTTTTTGTTGTTAT GAAAGGTCCAAATGATGCTTTGTTGAGATGGCCATTTAATCAAAAAGTTACTTTGATGTTGTTGGATCAAAATAA TAGAGAACATGTTATTGATGCTTTTAGACCAGATGTTACTTCTTCTTCTTTTCAAAGACCAGTTAATGATATGAAT ATTGCTTCTGGTTGTCCATTGTTTTGTCCAGTTTCTAAAATGGAAGCTAAAAATTCTTATGTTAGAGATGATGCTA TTTTTATTAAAGCTATTGTTGATTTGACTGGTTTGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTT CTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTT GGTTCTGCTTCTATTAAATTGCAATCTTCTGATGGTGAAATTTTTGAAGTTGATGTTGAAATTGCTAAACAATCTG TTACTATTAAAACTATGTTGGAAGATTTGGGTATGGATGATGAAGGTGATGATGATCCAGTTCCATTGCCAAATG TTAATGCTGCTATTTTGAAAAAAGTTATTCAATGGTGTACTCATCATAAAGATGATCCACCACCACCAGAAGATG ATGAAAATAAAGAAAAAAGAACTGATGATATTCCAGTTTGGGATCAAGAATTTTTGAAAGTTGATCAAGGTACTTT GTTTGAATTGATTTTGGCTGCTAATTATTTGGATATTAAAGGTTTGTTGGATGTTACTTGTAAAACTGTTGCTAAT ATGATTAAAGGTAAAACTCCAGAAGAAATTAGAAAAACTTTTAATATTAAAAATGATTTTACTGAAGAAGAAGAAG CTCAAGTTAGAAAAGAAAATCAATGGTGT

Figure 12B (continued)

Isopentenyl-Diphosphate Delta-lsomerase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID12

ATGACTGCCGACAACAATAGTATGCCCCATGGTGCAGTATCTAGTTACGCCAAATTAGTGCAAAACCAAACACC TGAAGACATTTTGGAAGAGTTTCCTGAAATTATTCCATTACAACAAAGACCTAATACCCGATCTAGTGAGACGTC AAATGACGAAAGCGGAGAAACATGTTTTTCTGGTCATGATGAGGAGCAAATTAAGTTAATGAATGAAAATTGTAT TGTTTTGGATTGGGACGATAATGCTATTGGTGCCGGTACCAAGAAAGTTTGTCATTTAATGGAAAATATTGAAAA GGGTTTACTACATCGTGCATTCTCCGTCTTTATTTTCAATGAACAAGGTGAATTACTTTTACAACAAAGAGCCAC TGAAAAAATAACTTTCCCTGATCTTTGGACTAACACATGCTGCTCTCATCCACTATGTATTGATGACGAATTAGG TTTGAAGGGTAAGCTAGACGATAAGATTAAGGGCGCTATTACTGCGGCGGTGAGAAAACTAGATCATGAATTAG GTATTCCAGAAGATGAAACTAAGACAAGGGGTAAGTTTCACTTTTTAAACAGAATCCATTACATGGCACCAAGCA ATGAACCATGGGGTGAACATGAAATTGATTACATCCTATTTTATAAGATCAACGCTAAAGAAAACTTGACTGTCA ACCCAAACGTCAATGAAGTTAGAGACTTCAAATGGGTTTCACCAAATGATTTGAAAACTATGTTTGCTGACCCAA GTTACAAGTTTACGCCTTGGTTTAAGATTATTTGCGAGAATTACTTATTCAACTGGTGGGAGCAATTAGATGACC TTTCTGAAGTGGAAAATGACAGGCAAATTCATAGAATGCTATAAAAATTGTCTGGTGGTGGTGGTTCTGGTGGT GGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTAT CAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGT AATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATA ATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCA AAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACA AGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTAT AATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGG ATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTG AAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGT GGTGGTTCTGGTGGTGGTGGTGCTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATA AAAAAGCTGGTTCTGGTTCTAATACTGTTAAAGAATTGAAAAATTATATTCAAGAATTGGAAGAAAGAAATGCTG AATTGAAAAATTTGAAAGAACATTTGAAATTTGCTAAAGCTGAATTGGAATTTGAATTGGCTGCTCATAAATTTGA AGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTA AAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTT GGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTGCTCAATTGAAAAATAGAGTTGCTTAT AAATTGAAAGAAAATGCTAAATTGGAAAATATTGTTGCTAGATTGGAAAATGATAATGCTAATTTGGAAAAAGATA TTGCTAATTTGGAAAAAGATATTGCTAATTTGGAAAGAGATGTTGCTAGA

Geranyl-Diphosphate Synthase (ERG20ww) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID13

ATGGCTTCAGAAAAGGAAATAAGAAGAGAAAGATTCTTGAACGTATTCCCAAAGTTAGTTGAAGAATTGAACGCT AGTTTGTTAGCTTATGGTATGCCTAAAGAAGCCTGCGATTGGTATGCTCACTCTTTAAACTACAATACTCCAGGT GGTAAATTGAATAGAGGTTTGAGTGTAGTTGATACTTATGCTATCTTGTCTAACAAAACCGTTGAACAATTAGGT CAAGAAGAATACGAAAAGGTCGCTATCTTGGGTTGGTGTATTGAATTGTTGCAAGCATACTTTTTGGTTGCCGAT GACATGATGGATAAGTCTATAACAAGAAGAGGTCAACCATGCTGGTACAAAGTTCCAGAAGTTGGTGAAATAGC CATAAATGATGCTTTTATGTTGGAAGCCGCTATCTATAAATTGTTGAAGTCACATTTCAGAAACGAAAAGTACTA CATCGATATTACCGAATTATTCCACGAAGTTACTTTCCAAACAGAATTGGGTCAATTGATGGATTTGATAACTGC ACCTGAAGATAAAGTTGACTTGTCAAAGTTTTCCTTGAAGAAACATTCATTCATCGTCACCTTTGAAACTGCTTAT TACTCCTTCTATTTGCCAGTCGCCTTGGCTATGTACGTAGCTGGTATTACTGATGAAAAAGACTTGAAGCAAGCA AGAGATGTTTTGATACCTTTGGGTGAATACTTCCAAATCCAAGATGACTACTTAGACTGTTTCGGTACTCCAGAA CAAATAGGTAAAATCGGTACAGATATTCAAGACAATAAGTGCAGTTGGGTTATTAACAAGGCTTTGGAATTAGCA TCTGCCGAACAAAGAAAGACTTTGGATGAAAACTACGGTAAAAAGGACTCAGTTGCTGAAGCAAAGTGTAAGAA AATTTTTAATGATTTGAAGATTGAACAATTGTACCATGAATACGAAGAATCCATCGCTAAAGACTTAAAGGCAAA GATTAGTCAAGTTGATGAATCAAGAGGTTTTAAAGCCGACGTTTTGACAGCTTTCTTGAATAAGGTCTACAAGAG ATCAAAGTAGAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCT TGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGG AATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAG CTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTA

Figure 12B (continued)

TTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAA GCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAA AAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTA TCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGG TAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAAT AATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTTGTGTA CTATGAAAAAAGGTCCATCTGGTTATGGTTTTAATTTGCATTCTGATAAATCTAAACCAGGTCAATTTATTAGATC TGTTGATCCAGATTCTCCAGCTGAAGCTTCTGGTTTGAGAGCTCAAGATAGAATTGTTGAAGTTAATGGTGTTTG TATGGAAGGTAAACAACATGGTGATGTTGTTTCTGCTATTAGAGCTGGTGGTGATGAAACTAAATTGTTGGTTGT TGATAGAGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAG CTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTCTGGTGCTATTATTT ATACTGTTGAATTGAAAAGATATGGTGGTCCATTGGGTATTACTATTTCTGGTACTGAAGAACCATTTGATCCAA TTATTATTTCTTCTTTGACTAAAGGTGGTTTGGCTGAAAGAACTGGTGCTATTCATATTGGTGATAGAATTTTGGC TATTAATTCTTCTTCTTTGAAAGGTAAACCATTGTCTGAAGCTATTCATTTGTTGCAAATGGCTGGTGAAACTGTT ACTTTGAAAATTAAAAAACAAACTGATGCTCAACCAGCTTCTTCT

Olivetol Synthase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID14

ATGAATCATTTGAGAGCTGAAGGTCCAGCTTCTGTTTTGGCTATTGGTACTGCTAATCCAGAAAATATTTTGTTG CAAGATGAATTTCCAGATTATTATTTTAGAGTTACTAAATCTGAACATATGACTCAATTGAAAGAAAAATTTAGAA AAATTTGTGATAAATCTATGATTAGAAAAAGAAATTGTTTTTTGAATGAAGAACATTTGAAACAAAATCCAAGATT GGTTGAACATGAAATGCAAACTTTGGATGCTAGACAAGATATGTTGGTTGTTGAAGTTCCAAAATTGGGTAAAGA TGCTTGTGCTAAAGCTATTAAAGAATGGGGTCAACCAAAATCTAAAATTACTCATTTGATTTTTACTTCTGCTTCT ACTACTGATATGCCAGGTGCTGATTATCATTGTGCTAAATTGTTGGGTTTGTCTCCATCTGTTAAAAGAGTTATG ATGTATCAATTGGGTTGTTATGGTGGTGGTACTGTTTTGAGAATTGCTAAAGATATTGCTGAAAATAATAAAGGT GCTAGAGTTTTGGCTGTTTGTTGTGATATTATGGCTTGTTTGTTTAGAGGTCCATCTGAATCTGATTTGGAATTG TTGGTTGGTCAAGCTATTTTTGGTGATGGTGCTGCTGCTGTTATTGTTGGTGCTGAACCAGATGAATCTGTTGG TGAAAGACCAATTTTTGAATTGGTTTCTACTGGTCAAACTATTTTGCCAAATTCTGAAGGTACTATTGGTGGTCAT ATTAGAGAAGCTGGTTTGATTTTTGATTTGCATAAAGATGTTCCAATGTTGATTTCTAATAATATTGAAAAATGTTT GATTGAAGCTTTTACTCCAATTGGTATTTCTGATTGGAATTCTATTTTTTGGATTACTCATCCAGGTGGTAAAGCT ATTTTGGATAAAGTTGAAGAAAAATTGCATTTGAAATCTGATAAATTTGTTGATTCTAGACATGTTTTGTCTGAAC ATGGTAATATGTCTTCTTCTACTGTTTTGTTTGTTATGGATGAATTGAGAAAAAGATCTTTGGAAGAAGGTAAATC TACTACTGGTGATGGTTTTGAATGGGGTGTTTTGTTTGGTTTTGGTCCAGGTTTGACTGTTGAAAGAGTTGTTGT TAGATCTGTTCCAATTAAATATAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTT CTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCA AAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGA TTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTG GGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAA ATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGA ATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAA CAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGG TATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAAT TGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGC TTCTGGTAATAATTTGGAAACTTATGAATGGTATAATAAATCTATTTCTAGAGATAAAGCTGAAAAATTGTTGTTG GAT ACTGGTAAAGAAGGTGCTTTTATGGTTAGAGATTCTAGAACTCCAGGTACTTATACTGTTTCTGTTTTT ACTA AAGCTATTATTTCTGAAAATCCATGTATTAAACATTATCATATTAAAGAAACTAATGATTCTCCAAAAAGATATTAT GTTGCTGAAAAATATGTTTTTGATTCTATTCCATTGTTGATTCAATATCATCAATATAATGGTGGTGGTTTGGTTA CTAGATTGAGATATCCAGTTTGTGGTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAA GCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGG TTCTCATCCATGGTTTTTTGGTAAAATTCCAAGAGCTAAAGCTGAAGAAATGTTGTCTAAACAAAGACATGATGG TGCTTTTTTGATTAGAGAATCTGAATCTGCTCCAGGTGATTTTTCTTTGTCTGTTAAATTTGGTAATGATGTTCAA

Figure 12B (continued)

CATTTTAAAGTTTTGAGAGATGGTGCTGGTAAATATTTTTTGTGGGTTGTTAAATTTAATTCTTTGAATGAATTGG TTGATTATCATAGATCTACTTCTGTTTCTAGAAATCAACAAATTTTTTTGAGAGATATTGAACAAGTTCCACAACA ACCAACT

Olivetolic Acid Cyclase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID15

ATGGCTGTTAAACATTTGATTGTTTTGAAATTTAAAGATGAAATTACTGAAGCTCAAAAAGAAGAATTTTTTAAAA CTTATGTTAATTTGGTTAATATTATTCCAGCTATGAAAGATGTTTATTGGGGTAAAGATGTTACTCAAAAAAATAA AGAAGAAGGTTATACTCATATTGTTGAAGTTACTTTTGAATCTGTTGAAACTATTCAAGATTATATTATTCATCCA GCTCATGTTGGTTTTGGTGATGTTTATAGATCTTTTTGGGAAAAATTGTTGATTTTTGATTATACTCCAAGAAAAA AATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTT GGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCA AATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATT ATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAG GTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTT GGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGA ATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCT ATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATT ATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAG ATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGGTCAAGATAGATCTGAA GCTACTTTGATTAAAAGATTTAAAGGTGAAGGTGTTAGATATAAAGCTAAATTGATTGGTATTGATGAAGTTTCTG CTGCTAGAGGTGATAAATTGTGTCAAGATTCTATGATGAAATTGAAAGGTGTTGTTGCTGGTGCTAGATCTAAAG GTGAACATAAACAAAAAATTTTTTTGACTATTTCTTTTGGTGGTATTAAAATTTTTGATGAAAAAACTGGTGCTTTG CAACATCATCATGCTGTTCATGAAATTTCTTATATTGCTAAAGATATTACTGATCATAGAGCTTTTGGTTATGTTT GTGGTAAAGAAGGTAATCATAGATTTGTTGCTATTAAAACTGCTCAAGCTGCTGAACCAGTTATTTTGGATTTGA GAGATTTGTTTCAATTGATTTATGAATTGAAACAAAGAGAAGAATTGGAAAAAAAAGCTGGTTCTGCTGGTTCTG CTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGT TCTGCTGCTGGTTCTGGTGAATTTGGTTCTGGTTCTCATATGGGTTCTCAATTTTGGGTTACTTCTCAAAAAACT GAAGCTTCTGAAAGATGTGGTTTGCAAGGTTCTTATATTTTGAGAGTTGAAGCTGAAAAATTGACTTTGTTGACT TTGGGTGCTCAATCTCAAATTTTGGAACCATTGTTGTTTTGGCCATATACTTTGTTGAGAAGATATGGTAGAGAT AAAGTTATGTTTTCTTTTGAAGCTGGTAGAAGATGTCCATCTGGTCCAGGTACTTTTACTTTTCAAACTTCTCAAG GTAATGATATTTTTCAAGCTGTTGAAGCTGCTATTCAACAACAAAAAGCTCAAGGTAAAGTTGGTCAAGCTCAAG ATATTTTGAGATTGGAACATCATCATCATCATCAT

CBGA Synthase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID16

ATGGGTTTGTCTTCTGTTTGTACTTTTTCTTTTCAAACTAATTATCATACTTTGTTGAATCCACATAATAATAATCC AAAAACTTCTTTGTTGTGTTATAGACATCCAAAAACTCCAATTAAATATTCTTATAATAATTTTCCATCTAAACATT GTTCTACTAAATCTTTTCATTTGCAAAATAAATGTTCTGAATCTTTGTCTATTGCTAAAAATTCTATTAGAGCTGCT ACTACTAATCAAACTGAACCACCAGAATCTGATAATCATTCTGTTGCTACTAAAATTTTGAATTTTGGTAAAGCTT GTTGGAAATTGCAAAGACCATATACTATTATTGCTTTTACTTCTTGTGCTTGTGGTTTGTTTGGTAAAGAATTGTT GCATAATACTAATTTGATTTCTTGGTCTTTGATGTTTAAAGCTTTTTTTTTTTTGGTTGCTATTTTGTGTATTGCTT CTTTTACTACTACTATTAATCAAATTTATGATTTGCATATTGATAGAATTAATAAACCAGATTTGCCATTGGCTTCT GGTGAAATTTCTGTTAATACTGCTTGGATTATGTCTATTATTGTTGCTTTGTTTGGTTTGATTATTACTATTAAAAT GAAAGGTGGTCCATTGTATATTTTTGGTTATTGTTTTGGTATTTTTGGTGGTATTGTTTATTCTGTTCCACCATTTA GATGGAAACAAAATCCATCTACTGCTTTTTTGTTGAATTTTTTGGCTCATATTATTACTAATTTTACTTTTTATTAT GCTTCTAGAGCTGCTTTGGGTTTGCCATTTGAATTGAGACCATCTTTTACTTTTTTGTTGGCTTTTATGAAATCTA TGGGTTCTGCTTTGGCTTTGATTAAAGATGCTTCTGATGTTGAAGGTGATACTAAATTTGGTATTTCTACTTTGG CTTCTAAATATGGTTCTAGAAATTTGACTTTGTTTTGTTCTGGTATTGTTTTGTTGTCTTATGTTGCTGCTATTTTG GCTGGTATTATTTGGCCACAAGCTTTTAATTCTAATGTTATGTTGTTGTCTCATGCTATTTTGGCTTTTTGGTTGA

Figure 12B (continued)

TTTTGCAAACTAGAGATTTTGCTTTGACTAATTATGATCCAGAAGCTGGTAGAAGATTTTATGAATTTATGTGGAA ATTGTATTATGCTGAATATTTGGTTTATGTTTTTATTAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCT GGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCT ATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATT ATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTGAAG CTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTT GGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAAATTTGGGTAATGCTTATTATAAACAAGGTGATTA TCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTTCTGCTTGGTATAATTTGGGT AATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATA ATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAAGATTATC AAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCT GGTGGTGGTGGTGCTTCTGCTGAATATGTTAGAGCTTTGTTTGATTTTAATGGTAATGATGAAGAAGATTTGCCA TTTAAAAAAGGTGATATTTTGAGAATTAGAGATAAACCAGAAGAACAATGGTGGAATGCTGAAGATTCTGAAGGT AAAAGAGGTATGATTCCAGTTCCATATGTTGAAAAATATGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTT GGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTG AATTTGGTTCTTTGATTAAACATATGAGAGCTGAAGCTTTGTTTGATTTTACTGGTAATTCTAAATTGGAATTGAA TTTTAAAGCTGGTGATGTTATTTTTTTGTTGTCTAGAATTAATAAAGATTGGTTGGAAGGTACTGTTAGAGGTGCT ACTGGTATTTTTCCATTGTCTTTTGTTAAAATTTTGAAA

Acetyl-CoA Carboxylase - Enzyme Linker - cTPR6 Spacer - ID Linker - ID17

ATGAGCGAAGAAAGCTTATTCGAGTCTTCTCCACAGAAGATGGAGTACGAAATTACAAACTACTCAGAAAGACA TACAGAACTTCCAGGTCATTTCATTGGCCTCAATACAGTAGATAAACTAGAGGAGTCCCCGTTAAGGGACTTTG TTAAGAGTCACGGTGGTCACACGGTCATATCCAAGATCCTGATAGCAAATAATGGTATTGCCGCCGTGAAAGAA ATTAGATCCGTCAGAAAATGGGCATACGAGACGTTCGGCGATGACAGAACCGTCCAATTCGTCGCCATGGCCA CCCCAGAAGATCTGGAGGCCAACGCAGAATATATCCGTATGGCCGATCAATACATTGAAGTGCCAGGTGGTAC TAATAATAACAACTACGCTAACGTAGACTTGATCGTAGACATCGCCGAAAGAGCAGACGTAGACGCCGTATGG GCTGGCTGGGGTCACGCCTCCGAGAATCCACTATTGCCTGAAAAATTGTCCCAGTCTAAGAGGAAAGTCATCTT TATTGGGCCTCCAGGTAACGCCATGAGGTCTTTAGGTGATAAAATCTCCTCTACCATTGTCGCTCAAAGTGCTA AAGTCCCATGTATTCCATGGTCTGGTACCGGTGTTGACACCGTTCACGTGGACGAGAAAACCGGTCTGGTCTC TGTCGACGATGACATCTATCAAAAGGGTTGTTGTACCTCTCCTGAAGATGGTTTACAAAAGGCCAAGCGTATTG GTTTTCCTGTCATGATTAAGGCATCCGAAGGTGGTGGTGGTAAAGGTATCAGACAAGTTGAACGTGAAGAAGAT TTCATCGCTTTATACCACCAGGCAGCCAACGAAATTCCAGGCTCCCCCATTTTCATCATGAAGTTGGCCGGTAG AGCGCGTCACTTGGAAGTTCAACTGCTAGCAGATCAGTACGGTACAAATATTTCCTTGTTCGGTAGAGACTGTT CCGTTCAGAGACGTCATCAAAAAATTATCGAAGAAGCACCAGTTACAATTGCCAAGGCTGAAACATTTCACGAG ATGGAAAAGGCTGCCGTCAGACTGGGGAAACTAGTCGGTTATGTCTCTGCCGGTACCGTGGAGTATCTATATT CTCATGATGATGGAAAATTCTACTTTTTAGAATTGAACCCAAGATTACAAGTCGAGCATCCAACAACGGAAATGG TCTCCGGTGTTAACTTACCTGCAGCTCAATTACAAATCGCTATGGGTATCCCTATGCATAGAATAAGTGACATTA GAACTTTATATGGTATGAATCCTCATTCTGCCTCAGAAATCGATTTCGAATTCAAAACTCAAGATGCCACCAAGA AACAAAGAAGACCTATTCCAAAGGGTCATTGTACCGCTTGTCGTATCACATCAGAAGATCCAAACGATGGATTC AAGCCATCGGGTGGTACTTTGCATGAACTAAACTTCCGTTCTTCCTCTAATGTTTGGGGTTACTTCTCCGTGGG TAACAATGGTAATATTCACTCCTTTTCGGACTCTCAGTTCGGCCATATTTTTGCTTTTGGTGAAAATAGACAAGCT TCCAGGAAACACATGGTTGTTGCCCTGAAGGAATTGTCCATTAGGGGTGATTTCAGAACTACTGTGGAATACTT GATCAAACTTTTGGAAACTGAAGATTTCGAGGATAACACTATTACCACCGGTTGGTTGGACGATTTGATTACTCA TAAAATGACCGCTGAAAAGCCTGATCCAACTCTTGCCGTCATTTGCGGTGCCGCTACAAAGGCTTTCTTAGCAT CTGAAGAAGCCCGCCACAAGTATATCGAATCCTTACAAAAGGGACAAGTTCTATCTAAAGACCTACTGCAAACT ATGTTCCCTGTAGATTTTATCCATGAGGGTAAAAGATACAAGTTCACCGTAGCTAAATCCGGTAATGACCGTTAC ACATTATTTATCAATGGTTCTAAATGTGATATCATACTGCGTCAACTATCTGATGGTGGTCTTTTGATTGCCATAG GCGGTAAATCGCATACCATCTATTGGAAAGAAGAAGTTGCTGCTACAAGATTATCCGTTGACTCTATGACTACTT TGTTGGAAGTTGAAAACGATCCAACCCAGTTGCGTACTCCATCCCCTGGTAAATTGGTTAAATTCTTGGTGGAA

Figure 12B (continued)

AATGGTGAACACATTATCAAGGGCCAACCATATGCAGAAATTGAAGTTATGAAAATGCAAATGCCTTTGGTTTCT CAAGAAAATGGTATCGTCCAGTTATTAAAGCAACCTGGTTCTACCATTGTTGCAGGTGATATCATGGCTATTATG ACTCTTGACGATCCATCCAAGGTCAAGCACGCTCTACCATTTGAAGGTATGCTGCCAGATTTTGGTTCTCCAGT TATCGAAGGAACCAAACCTGCCTATAAATTCAAGTCATTAGTGTCTACTTTGGAAAACATTTTGAAGGGTTATGA CAACCAAGTTATTATGAACGCTTCCTTGCAACAATTGATAGAGGTTTTGAGAAATCCAAAACTGCCTTACTCAGA ATGGAAACTACACATCTCTGCTTTACATTCAAGATTGCCTGCTAAGCTAGATGAACAAATGGAAGAGTTAGTTGC ACGTTCTTTGAGACGTGGTGCTGTTTTCCCAGCTAGACAATTAAGTAAATTGATTGATATGGCCGTGAAGAATCC TGAATACAACCCCGACAAATTGCTGGGCGCCGTCGTGGAACCATTGGCGGATATTGCTCATAAGTACTCTAAC GGGTTAGAAGCCCATGAACATTCTATATTTGTCCATTTCTTGGAAGAATATTACGAAGTTGAAAAGTTATTCAAT GGTCCAAATGTTCGTGAGGAAAATATCATTCTGAAATTGCGTGATGAAAACCCTAAAGATCTAGATAAAGTTGCG CTAACTGTTTTGTCTCATTCGAAAGTTTCAGCGAAGAATAACCTGATCCTAGCTATCTTGAAACATTATCAACCAT TGTGCAAGTTATCTTCTAAAGTTTCTGCCATTTTCTCTACTCCTCTACAACATATTGTTGAACTAGAATCTAAGGC TACCGCTAAGGTCGCTCTACAAGCAAGAGAAATTTTGATTCAAGGCGCTTTACCTTCGGTCAAGGAAAGAACTG AACAAATTGAACATATCTTAAAATCCTCTGTTGTGAAGGTTGCCTATGGCTCATCCAATCCAAAGCGCTCTGAAC CAGATTTGAATATCTTGAAGGACTTGATCGATTCTAATTACGTTGTGTTCGATGTTTTACTTCAATTCCTAACCCA TCAAGACCCAGTTGTGACTGCTGCAGCTGCTCAAGTCTATATTCGTCGTGCTTATCGTGCTTACACCATAGGAG ATATTAGAGTTCACGAAGGTGTCACAGTTCCAATTGTTGAATGGAAATTCCAACTACCTTCAGCTGCGTTCTCCA CCTTTCCAACTGTTAAATCTAAAATGGGTATGAACAGGGCTGTTTCTGTTTCAGATTTGTCATATGTTGCAAACA GTCAGTCATCTCCGTTAAGAGAAGGTATTTTGATGGCTGTGGATCATTTAGATGATGTTGATGAAATTTTGTCAC AAAGTTTGGAAGTTATTCCTCGTCACCAATCTTCTTCTAACGGACCTGCTCCTGATCGTTCTGGTAGCTCCGCAT CGTTGAGTAATGTTGCTAATGTTTGTGTTGCTTCTACAGAAGGTTTCGAATCTGAAGAGGAAATTTTGGTAAGGT TGAGAGAAATTTTGGATTTGAATAAGCAGGAATTAATCAATGCTTCTATCCGTCGTATCACATTTATGTTCGGTTT TAAAGATGGGTCTTATCCAAAGTATTATACTTTTAACGGTCCAAATTATAACGAAAATGAAACAATTCGTCACATT GAGCCGGCTTTGGCCTTCCAACTGGAATTAGGAAGATTGTCCAACTTCAACATTAAACCAATTTTCACTGATAAT AGAAACATCCATGTCTACGAAGCTGTTAGTAAGACTTCTCCATTGGATAAGAGATTCTTTACAAGAGGTATTATT AGAACGGGTCATATCCGTGATGACATTTCTATTCAAGAATATCTGACTTCTGAAGCTAACAGATTGATGAGTGAT ATATTGGATAATTTAGAAGTCACCGACACTTCAAATTCTGATTTGAATCATATCTTCATCAACTTCATTGCGGTGT TTGATATCTCTCCAGAAGATGTCGAAGCCGCCTTCGGTGGTTTCTTAGAAAGATTTGGTAAGAGATTGTTGAGA TTGCGTGTTTCTTCTGCCGAAATTAGAATCATCATCAAAGATCCTCAAACAGGTGCCCCAGTACCATTGCGTGC CTTGATCAATAACGTTTCTGGTTATGTTATCAAAACAGAAATGTACACCGAAGTCAAGAACGCAAAAGGTGAATG GGTATTTAAGTCTTTGGGTAAACCTGGATCCATGCATTTAAGACCTATTGCTACTCCTTACCCTGTTAAGGAATG GTTGCAACCAAAACGTTATAAGGCACACTTGATGGGTACCACATATGTCTATGACTTCCCAGAATTATTCCGCCA AGCATCGTCATCCCAATGGAAAAATTTCTCTGCAGATGTTAAGTTAACAGATGATTTCTTTATTTCCAACGAGTT GATTGAAGATGAAAACGGCGAATTAACTGAGGTGGAAAGAGAACCTGGTGCCAACGCTATTGGTATGGTTGCC TTTAAGATTACTGTAAAGACTCCTGAATATCCAAGAGGCCGTCAATTTGTTGTTGTTGCTAACGATATCACATTC AAGATCGGTTCCTTTGGTCCACAAGAAGACGAATTCTTCAATAAGGTTACTGAATATGCTAGAAAGCGTGGTAT CCCAAGAATTTACTTGGCTGCAAACTCAGGTGCCAGAATTGGTATGGCTGAAGAGATTGTTCCACTATTTCAAG TTGCATGGAATGATGCTGCCAATCCGGACAAGGGCTTCCAATACTTATACTTAACAAGTGAAGGTATGGAAACT TTAAAGAAATTTGACAAAGAAAATTCTGTTCTCACTGAACGTACTGTTATAAACGGTGAAGAAAGATTTGTCATCA AGACAATTATTGGTTCTGAAGATGGGTTAGGTGTCGAATGTCTACGTGGATCTGGTTTAATTGCTGGTGCAACG TCAAGGGCTTACCACGATATCTTCACTATCACCTTAGTCACTTGTAGATCCGTCGGTATCGGTGCTTATTTGGTT CGTTTGGGTCAAAGAGCTATTCAGGTCGAAGGCCAGCCAATTATTTTAACTGGTGCTCCTGCAATCAACAAAAT GCTGGGTAGAGAAGTTTATACTTCTAACTTACAATTGGGTGGTACTCAAATCATGTATAACAACGGTGTTTCACA TTTGACTGCTGTTGACGATTTAGCTGGTGTAGAGAAGATTGTTGAATGGATGTCTTATGTTCCAGCCAAGCGTA ATATGCCAGTTCCTATCTTGGAAACTAAAGACACATGGGATAGACCAGTTGATTTCACTCCAACTAATGATGAAA CTTACGATGTAAGATGGATGATTGAAGGTCGTGAGACTGAAAGTGGATTTGAATATGGTTTGTTTGATAAAGGG TCTTTCTTTGAAACTTTGTCAGGATGGGCCAAAGGTGTTGTCGTTGGTAGAGCCCGTCTTGGTGGTATTCCACT GGGTGTTATTGGTGTTGAAACAAGAACTGTCGAGAACTTGATTCCTGCTGATCCAGCTAATCCAAATAGTGCTG AAACATTAATTCAAGAACCTGGTCAAGTTTGGCATCCAAACTCCGCCTTCAAGACTGCTCAAGCTATCAATGACT TTAACAACGGTGAACAATTGCCAATGATGATTTTGGCCAACTGGAGAGGTTTCTCTGGTGGTCAACGTGATATG TTCAACGAAGTCTTGAAGTATGGTTCGTTTATTGTTGACGCATTGGTGGATTACAAACAACCAATTATTATCTATA TCCCACCTACCGGTGAACTAAGAGGTGGTTCATGGGTTGTTGTCGATCCAACTATCAACGCTGACCAAATGGAA

Figure 12B (continued)

ATGTATGCCGACGTCAACGCTAGAGCTGGTGTTTTGGAACCACAAGGTATGGTTGGTATCAAGTTCCGTAGAGA AAAATTGCTGGACACCATGAACAGATTGGATGACAAGTACAGAGAATTGAGATCTCAATTATCCAACAAGAGTTT GGCTCCAGAAGTACATCAGCAAATATCCAAGCAATTAGCTGATCGTGAGAGAGAACTATTGCCAATTTACGGAC AAATCAGTCTTCAATTTGCTGATTTGCACGATAGGTCTTCACGTATGGTGGCCAAGGGTGTTATTTCTAAGGAAC TGGAATGGACCGAGGCACGTCGTTTCTTCTTCTGGAGATTGAGAAGAAGATTGAACGAAGAATATTTGATTAAA AGGTTGAGCCATCAGGTAGGCGAAGCATCAAGATTAGAAAAGATCGCAAGAATTAGATCGTGGTACCCTGCTT CAGTGGACCATGAAGATGATAGGCAAGTCGCAACATGGATTGAAGAAAACTACAAAACTTTGGACGATAAACTA AAGGGTTTGAAATTAGAGTCATTCGCTCAAGACTTAGCTAAAAAGATCAGAAGCGACCATGACAATGCTATTGAT GGATTATCTGAAGTTATCAAGATGTTATCTACCGATGATAAAGAAAAATTGTTGAAGACTTTGAAATAAAAATTGT CTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGCTGAAGCTTGGTATAATTTGGGTAA TGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAAT GCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAA AAGCTTTGGAATTGGATCCAAATAATGCTGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATT ATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATTTGCAAGCTGAAGCTTGGAAAA ATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAATATTATCAAAAAGCTTTGGAATTGGA TCCAAATAATGCTTCTGCTTGGTATAATTTGGGTAATGCTTATTATAAACAAGGTGATTATCAAAAAGCTATTGAA TATTATCAAAAAGCTTTGGAATTGGATCCAAATAATGCTAAAGCTTGGTATAGAAGAGGTAATGCTTATTATAAAC AAGGTGATTATCAAAAAGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAATAATAGATCTAGATCTG CTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGGTTCTCATATGAGATTGGGTGC TCAATCTATTCAACCAACTGCTAATTTGGATAGAACTGATGATTTGGTTTATTTGAATGTTATGGAATTGGTTAGA GCTGTTTTGGAATTGAAAAATGAATTGGCTCAATTGCCACCAGAAGGTTATGTTGTTGTTGTTAAAAATGTTGGT TTGACTTTGAGAAAATTGATTGGTTCTGTTGATGATTTGTTGCCATCTTTGCCATCTTCTTCTAGAACTGAAATTG AAGGTACTCAAAAATTGTTGAATAAAGATTTGGCTGAATTGATTAATAAAATGAGATTGGCTCAACAAAATGCTG TTACTTCTTTGTCTGAAGAATGTAAAAGACAAATGTTGACTGCTTCTCATACTTTGGCTGTTGATGCTAAAAATTT GTTGGATGCTGTTGATCAAGCTAAAGTTTTGGCTAATTTGGCTCATCCACCAGCTGAAGGTTCTGCTGGTTCTG CTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGT TCTGCTGCTGGTTCTGGTGAATTTGGTTCTGGTGCTATGGCTACTCCAGGTTCTGAAAATGTTTTGCCAAGAGA ACCATTGATTGCTACTGCTGTTAAATTTTTGCAAAATTCTAGAGTTAGACAATCTCCATTGGCTACTAGAAGAGC TTTTTTGAAAAAAAAAGGTTTGACTGATGAAGAAATTGATATGGCTTTTCAACAATCTGGTACTGCTGCTGATGA ACCATCTTCTTTGTGG

Figure 12C

Cannabinoidergic Metabolon Scaffold - (Myc)3

ATGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTG AATTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGG AATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTGCTAGAAATGCTTATTTGAGAAAAAAAATTGCTA GATTGAAAAAAGATAATTTGCAATTGGAAAGAGATGAACAAAATTTGGAAAAAATTATTGCTAATTTGAGAG ATGAAATTGCTAGATTGGAAAATGAAGTTGCTTCTCATGAACAAGGTTCTGCTGGTTCTGCTGCTGGTTCT GGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTT CTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTC TGGTTCTAATTTGGTTGCTCAATTGGAAAATGAAGTTGCTTCTTTGGAAAATGAAAATGAAACTTTGAAAAA AAAAAATTTGCATAAAAAAGATTTGATTGCTTATTTGGAAAAAGAAATTGCTAATTTGAGAAAAAAAATTGAA GAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTG CTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGG TGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGT TCTGGTTCTCAAAAAGTTGCTGAATTGAAAAATAGAGTTGCTGTTAAATTGAATAGAAATGAACAATTGAAA AATAAAGTTGAAGAATTGAAAAATAGAAATGCTTATTTGAAAAATGAATTGGCTACTTTGGAAAATGAAGTT GCTAGATTGGAAAATGATGTTGCTGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGC TGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTATT ATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTAATGAAGTTAC TACTTTGGAAAATGATGCTGCTTTTATTGAAAATGAAAATGCTTATTTGGAAAAAGAAATTGCTAGATTGAG AAAAGAAAAAGCTGCTTTGAGAAATAGATTGGCTCATAAAAAAGGTTCTGCTGGTTCTGCTGCTGGTTCTG GTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGC TGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTAGACCACCAACTATTTCTA ATCCACCACCATTGATTTCTTCTGCTAAACATCCATCTGTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGT GAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTG GTGAATTTAATTTTTTGCAATCTAGACCAGAACCAACTGCTCCACCAGAAGAATCTTTTAGATCTGGTGGTT CTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAA AGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTT GGTTCTTCTAAAGGTACTGGTTTGAATCCAAATGCTAAAGTTTGGCAAGAAATTGCTCCAGGTAATGGTTC TGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGT TCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTCCAGATGGTGGTACTACTTTTGAACATTTGTGGTCTTC TTTGGAACCAGATTCTACTTATGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAG CTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGG TTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCT TTGTATAAAAAAGCTGGTTCTGGTTCTAAAAGAATTGCTTATTTGAGAAAAAAAATTGCTGCTTTGAAAAAA GATAATGCTAATTTGGAAAAAGATATTGCTAATTTGGAAAATGAAATTGAAAGATTGATTAAAGAAATTAAAA CTTTGGAAAATGAAGTTGCTTCTCATGAACAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCT GAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTT CTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTAATTT GTTGGCTACTTTGAGATCTACTGCTGCTGTTTTGGAAAATGAAAATCATGTTTTGGAAAAAGAAAAAGAAAA ATTGAGAAAAGAAAAAGAACAATTGTTGAATAAATTGGAAGCTTATAAAGGTTCTGCTGGTTCTGCTGCTG GTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGA AGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTCCAGCTACTTCTC AACATCCACCACCACCACCAGGTCATAGATCTCAAGCTCCATCTCATGGTTCTGCTGGTTCTGCTGCTGGT TCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTG GTTCTGGTGAATTTGAATTGAATTCTTTGTTGATTTTGTTGGAAGCTGCTGAATATTTGGAAAGAAGAGATA GAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGC TGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGT GAATTTGGTTCTAGACCACCAACTATTTCTAATCCACCACCATTGATTTCTTCTGCTAAACATCCATCTGTT GGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAG CTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTAATTTTTTGCAATCTAGACCAGAACCAACTGCT

Figure 12C (continued)

CCACCAGAAGAATCTTTTAGATCTGGTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGC TGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCT GCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTAAAGGTACTGGTTTGAATCCAAATGCTAAAGT TTGGCAAGAAATTGCTCCAGGTAATGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTG CTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTCCAGATGG TGGTACTACTTTTGAACATTTGTGGTCTTCTTTGGAACCAGATTCTACTTATGGTTCTGCTGGTTCTGCTGC TGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAA GAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCA TCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTAAAAGAATTGCTTAT TTGAGAAAAAAAATTGCTGCTTTGAAAAAAGATAATGCTAATTTGGAAAAAGATATTGCTAATTTGGAAAAT GAAATTGAAAGATTGATTAAAGAAATTAAAACTTTGGAAAATGAAGTTGCTTCTCATGAACAAGGTTCTGCT GGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTG CTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTT GTATAAAAAAGCTGGTTCTGGTTCTAATTTGTTGGCTACTTTGAGATCTACTGCTGCTGTTTTGGAAAATGA AAATCATGTTTTGGAAAAAGAAAAAGAAAAATTGAGAAAAGAAAAAGAACAATTGTTGAATAAATTGGAAGC TTATAAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAG CTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTC TGGTGAATTTGGTTCTGCTTTGGTTGATGATGCTGCTGATTATGAACCACCACCATCTAATAATGAAGAAG CTTTGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGC TAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTAGAGAATTGTTTGATGATCCATCTTATGT TAATGTTCAAAATTTGGATAAAGCTAGACAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTT CTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGG TTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTAAAAATACTAAATCTATGAATTTTGATAATCC AGTTTATAGAAAAACTACTGAAGAAGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAG CTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTAGATCT TTGCCATCTACTTGGATTGAAAATAAATTGTATGGTATGTCTGATCCAAATTGGGGTTCTGCTGGTTCTGCT GCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTA AAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGTTGTTGAT AATTCTCCACCACCAGCTTTGCCACCAAAAAAAAGACAATCTGCTCCATCTGGTTCTGCTGGTTCTGCTGC TGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCT GCTGGTTCTGGTGAATTTACTCAAAGATCTAAACCACAACCAGCTGTTCCACCAAGACCATCTGCTGATTT GATTTTGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAG CTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTC TGGTGAATTTGGTTCTACTGATGAAGAAAGAGAAGAAACTGAAGAAGAAGTTTATTTGTTGAATTCTACTAC TTTGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTA AAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGATGGTAATGTTTCTGGTACTCAAAGATTG GATTCTGCTACTGTTAGAACTTATTCTTGTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCT GCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTT CTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTTGGAAT CTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTGCTCAATTGAAAAATAGAGTTGCTTATA AATTGAAAGAAAATGCTAAATTGGAAAATATTGTTGCTAGATTGGAAAATGATAATGCTAATTTGGAAAAAG ATATTGCTAATTTGGAAAAAGATATTGCTAATTTGGAAAGAGATGTTGCTAGAGGTTCTGCTGGTTCTGCTG CTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGC TGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAA GCTGGTTCTGGTTCTAATACTGTTAAAGAATTGAAAAATTATATTCAAGAATTGGAAGAAAGAAATGCTGAA TTGAAAAATTTGAAAGAACATTTGAAATTTGCTAAAGCTGAATTGGAATTTGAATTGGCTGCTCATAAATTT GAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTG CTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGG TGAATTTGGTTCTCATGATGATTCTTTGCCACATCCACAACAAGCTACTGATGATTCTGGTCATGAATCTGA TGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAA

Figure 12C (continued)

GCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTCCAAATGCTGGTTCTGTTGAACAAAC TCCAAAAAAACCAGGTTTGAGAAGAAGAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTG CTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTC TGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTTGGAATC TACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTTTTGAAAATGTTACTCATGAATTTATTTTGGCTACTTTG GAAAATGAAAATGCTAAATTGAGAAGATTGGAAGCTAAATTGGAAAGAGAATTGGCTAGATTGAGAAATGA AGTTGCTTGGTTGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAA GCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCAT CATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTGAAGAATTGAAAAAT AAAATTGCTGAATTGGAAAATAGAAATGCTGTTAAAAAAAATAGAGTTGCTCATTTGAAACAAGAAATTGCT TATTTGAAAGATGAATTGGCTGCTCATGAATTTGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATT TGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAA GCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGTTTCTTCTACTAAATTGGTTTCTTTT CATGATGATTCTGATGAAGATTTGTTGCATATTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCT GAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTG CTGCTGCTACTCCAATTTCTACTTTTCATGATGATTCTGATGAAGATTTGTTGCATGTTGGTTCTGCTGGTT CTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGC TGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTT ATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGT TGAATCTTTGAAACAAAAAATTGAAGAATTGAAACAAAGAAAAGCTCAATTGAAAAATGATATTGCTAATTT GGAAAAAGAAATTGCTTATGCTGAAACTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAG CTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTAT TATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGAATTTTTTAGAAGAG AAAGAAATAAAATGGCTGCTGCTAAATGTAGAAATAGAAGAAGAGAATTGACTGATACTTTGCAAGCTGAA ACTGATCAATTGGAAGATGAAAAATCTGCTTTGCAAACTGAAATTGCTAATTTGTTGAAAGAAAAAGAAAAA TTGGAATTTATTTTGGCTGCTCATAGACCAGCTTGTAAAATTCCAGATGATTTGGGTTTTCCAGAAGAAATG TCTTTGGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGA AGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGT TCTGGTGAATTTGGTTCTTTTCAAATGCCAGCTGATACTCCACCACCAGCTTATTTGCCACCAGAAGATCC AATGACTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCT GCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGAAAGAGAATCTAATGAAGAACCACC ACCACCATATGAAGATCCATATTGGGGTAATGGTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTG GTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGC TGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTTCTTATTATCATCATCATCATCATCATTT GGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTCAAAAAGTTGCTGAATTGAAAAATAGAGTTGC TGTTAAATTGAATAGAAATGAACAATTGAAAAATAAAGTTGAAGAATTGAAAAATAGAAATGCTTATTTGAAA AATGAATTGGCTACTTTGGAAAATGAAGTTGCTAGATTGGAAAATGATGTTGCTGAAGGTTCTGCTGGTTC TGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGT TCTGCTGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATA AAAAAGCTGGTTCTGGTTCTAATGAAGTTACTACTTTGGAAAATGATGCTGCTTTTATTGAAAATGAAAATG CTTATTTGGAAAAAGAAATTGCTAGATTGAGAAAAGAAAAAGCTGCTTTGAGAAATAGATTGGCTCATAAAA AATCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTGC TAGAAATGCTTATTTGAGAAAAAAAATTGCTAGATTGAAAAAAGATAATTTGCAATTGGAAAGAGATGAACA AAATTTGGAAAAAATTATTGCTAATTTGAGAGATGAAATTGCTAGATTGGAAAATGAAGTTGCTTCTCATGA ACAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCT AAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTG GAATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTAATTTGGTTGCTCAATTGGAAAATGAAGTTGCT TCTTTGGAAAATGAAAATGAAACTTTGAAAAAAAAAAATTTGCATAAAAAAGATTTGATTGCTTATTTGGAAA AAGAAATTGCTAATTTGAGAAAAAAAATTGAAGAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTT GGTTCTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAG

Figure 12C (continued)

CTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGAACAAAAATTGATTTCTGAAGAAGAT TTGGAACAAAAATTGATTTCTGAAGAAGATTTGGAACAAAAATTGATTTCTGAAGAAGATTTGGGTTCTGCT GGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGC TGGTTCTGCTGCTGGTTCTGGTGAATTT

Figure 12D

Malonyl-CoA Metabolon Scaffold - (FLAG)3

ATGGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTG AATTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGG AATCTACTTCTTTGTATAAAAAAGCTGGTTCTGGTTCTGCTAGAAATGCTTATTTGAGAAAAAAAATTGCTA GATTGAAAAAAGATAATTTGCAATTGGAAAGAGATGAACAAAATTTGGAAAAAATTATTGCTAATTTGAGAG ATGAAATTGCTAGATTGGAAAATGAAGTTGCTTCTCATGAACAAGGTTCTGCTGGTTCTGCTGCTGGTTCT GGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTT CTGGTGAATTTTCTTATTATCATCATCATCATCATCATTTGGAATCTACTTCTTTGTATAAAAAAGCTGGTTC TGGTTCTAATTTGGTTGCTCAATTGGAAAATGAAGTTGCTTCTTTGGAAAATGAAAATGAAACTTTGAAAAA AAAAAATTTGCATAAAAAAGATTTGATTGCTTATTTGGAAAAAGAAATTGCTAATTTGAGAAAAAAAATTGAA GAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGAAGCTGCTGCTAAAGAAGCTG CTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGG TGAATTTGGTTCTTCTGCTACTAGAGAATTGGATGAATTGATGGCTTCTTTGTCTGATTTTAAAATTCAAGG TGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGCTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAA GCTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGATTTGGCTTTGTCTGAAAATTGGGCTCAAGA ATTTTTGGCTGCTGGTGATGCTGTTGATGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTG CTGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGAAGCTGCTGCTAAAGCTGGTTC TGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGATTATAAAGATGATGATGATAAAGATTATAAAGA TGATGATGATAAAGATTATAAAGATGATGATGATAAAGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAAT TTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAATTTGGTTCTGCTGGTTCTGCTGCTGGTTCTGGTGAA TTT

FIGURE 13 A

HCA Gene Cassette Amino Acid Sequences

1. ATP Citrate Lyase (ACL) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID1 (SEQ ID NQ:160)

MSAKAISEQTGKELLYKFICTTSAIQNRFKYARVTPDTDWARLLQDHPWLLSQNLWKPDQLIKRRGK LGLVGVNLTLDGVKSWLKPRLGQEATVGKATGFLKNFLIEPFVPHSQAEEFYVCIYATREGDYVLFHH EGGVDVGDVDAKAQKLLVGVDEKLNPEDIKKHLLVHAPEDKKEILASFISGLFNFYEDLYFTYLEINPL WTKDGVYVLDLAAKVDATADYICKVKWGDIEFPPPFGREAYPEEAYIADLDAKSGASLKLTLLNPKG RIWTMVAGGGASVVYSDTICDLGGVNELANYGEYSGAPSEQQTYDYAKTILSLMTREKHPDGKILIIG GSIANFTNVAATFKGIVRAIRDYQGPLKEHEVTIFVRRGGPNYQEGLRVMGEVGKTTGIPIHVFGTET HMTAIVGMALGHRPIPNQPPTAAHTANFLLNASGSTSTPAPSRTASFSESRADEVAPAKKAKPAMPQ DSVPSPRSLQGKSTTLFSRHTKAIVWGMQTRAVQGMLDFDYVCSRDEPSVAAMVYPFTGDHKQKF YWGHKEILIPVFKNMADAMRKHPEVDVLINFASLRSAYDSTMETMNYAQIRTIAIIAEGIPEALTRKLIKK ADQKGVTIIGPATVGGIKPGCFKIGNTGGMLDNILASKLYRPGSVAYVSRSGGMSNELNNIISRTTDGV YEGVAIGGDRYPGSTFMDHVLRYQDTPGVKMIWLGEIGGTEEYKICRGIKEGRLTKPIVCWCIGTCA TMFSSEVQFGHAGACANQASETAVAKNQALKEAGVFVPRSFDELGEIIQSVYEDLVANGVIVPAQEV PPPTVPMDYSWARELGLIRKPASFMTSICDERGQELIYAGMPITEVFKEEMGIGGVLGLLWFQKRLPK YSCQFIEMCLMVTADHGPAVSGAHNTIICARAGKDLVSSLTSGLLTIGDRFGGALDAAAKMFSKAFDS GIIPMEFVNKMKKEGKLIMGIGHRVKSINNPDMRVQILKDYVRQHFPATPLLDYALEVEKITTSKKPNLI LNVDGLIGVAFVDMLRNCGSFTREEADEYIDIGALNGIFVLGRSMGFIGHYLDQKRLKQGLYRHPWD DISYVLPEHMSMKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNA EAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPN NLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALE LDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASSYYH HHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENETLKKKNLHKKDLIAYLEKEIANLRKKIEEGSA GSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSARNAY LRKKIARLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASHEQGSG

2. Acetyl-CoA Acetyltransferase (atoB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID2 (SEQ ID

NO:161)

MKNCVIVSAVRTAIGSFNGSLASTSAIDLGATVIKAAIERAKIDSQHVDEVIMGNVLQAGLGQNPARQA LLKSGLAETVCGFTVNKVCGSGLKSVALAAQAIQAGQAQSIVAGGMENMSLAPYLLDAKARSGYRLG DGQVYDVILRDGLMCATHGYHMGITAENVAKEYGITREMQDELALHSQRKAAAAIESGAFTAEIVPVN WTRKKTFVFSQDEFPKANSTAEALGALRPAFDKAGTVTAGNASGINDGAAALVIMEESAALAAGLTP LARIKSYASGGVPPALMGMGPVPATQKALQLAGLQLADIDLIEANEAFAAQFLAVGKNLGFDSEKVNV NGGAIALGHPIGASGARILVTLLHAMQARDKTLGLATLCIGGGQGIAMVIERLNKLSGGGGSGGGGS GGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQK ALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIE YYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQK AIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSNEVTTL ENDAAFIENENAYLEKEIARLRKEKAALRNRLAHKKGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGS AAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVAELKNRVAVKLNRNEQLKNKVEELKNRNAYL KNELATLENEVARLENDVAEGSG

3. 3-Hvdroxvbutvryl-CoA Dehydrogenase (BHBD) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID3 (SEQ ID NO:162)

MKKVCVIGAGTMGSGIAQAFAAKGFEVVLRDIKDEFVDRGLDFINKNLSKLVKKGKIEEATKVEILTRIS GTVDLNMAADCDLVIEAAVERMDIKKQIFADLDNICKPETILASNTSSLSITEVASATKRPDKVIGMHFF NPAPVMKLVEVIRGIATSQETFDAVKETSIAIGKDPVEVAEAPGFWNRILIPMINEAVGILAEGIASVEDI DKAMKLGANHPMGPLELGDFIGLDICLAIMDVLYSETGDSKYRPHTLLKKYVRAGWLGRKSGKGFYD

YSKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGN AYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWK NLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKA WYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASENLYFQGENLYF QGDSSESCWNCGRKASETCSGCNTARYCGSFCQHKDWEKHHHICGQTLQAQQGSAGSAAGSGE FGSAEAAAKEAAAKAGSAGSAAGSGEFGSMAVSESQLKKMVSKYKYRDLTVRETVNVITLYKDLKPV LDSYVFNDGSSRELMNLTGTIPVPYRGNTYNIPICLWLLDTYPYNPPICFVKPTSSMTIKTGKHVDANG KIYLPYLHEWKHPQSDLLGLIQVMIWFGDEPPVFSRPGSG

4. Enoyl-CoA Hydratase (ECH) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID4 (SEQ ID NO:163)

MELNNVILEKEGKVAWTINRPKALNALNSDTLKEMDYVIGEIENDSEVLAVILTGAGEKSFVAGADISE MKEMNTIEGRKFGILGNKVFRRLELLEKPVIAAVNGFALGGGCEIAMSCDIRIASSNARFGQPEVGLGI TPGFGGTQRLSRLVGMGMAKQLIFTAQNIKADEALRIGLVNKVVEPSELMNTAKEIANKIVSNAPVAV KLSKQAINRGMQCDIDTALAFESEAFGECFSTEDQKDAMTAFIEKRKIEGFKNRKLSGGGGSGGGGS GGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQK ALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIE YYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQK AIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASGPLGSPLTASMLASAPPQEQKQMLGERLF PLIQAMHPTLAGKITGMLLEIDNSELLHMLESPESLRSKVDEAVAVLQAHQAKEAAQKAGSAGSAAGS GEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSNTNMSVPTDGAVTTSQIPASEQETLVRPKPLLLKLL KSVGAQKDTYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRN LWGSG

5. Trans-Enovl-CoA Reductase (ECR) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID5 (SEQ ID

NO: 164)

MIVKPMVRNNICLNAHPQGCKKGVEDQIEYTKKRITAEVKAGAKAPKNVLVLGCSNGYGLASRITAAF GYGAATIGVSFEKAGSETKYGTPGWYNNLAFDEAAKREGLYSVTIDGDAFSDEIKAQVIEEAKKKGIK FDLIVYSLASPVRTDPDTGIMHKSVLKPFGKTFTGKTVDPFTGELKEISAEPANDEEAAATVKVMGGE DWERWIKQLSKEGLLEEGCITLAYSYIGPEATQALYRKGTIGKAKEHLEATAHRLNKENPSIRAFVSVN KGLVTRASAVIPVIPLYLASLFKVMKEKGNHEGCIEQITRLYAERLYRKDGTIPVDEENRIRIDDWELEE DVQKAVSALMEKVTGENAESLTDLAGYRHDFLASNGFDVEGINYEAEVERFDRIKLSGGGGSGGGG SGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQ KALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAI EYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQ KAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSNLLAT LRSTAAVLENENHVLEKEKEKLRKEKEQLLNKLEAYKGSAGSAAGSGEFGSAEAAAKEAAAKAGSA GSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSKRIAYLRKKIAALKKDNANLEKDIANLENEIERLI KEIKTLENEVASHEQGSG

6. Beta-Ketothiolase (bktB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID6 (SEQ ID NO: 165)

MTREVWVSGVRTAIGTFGGSLKDVAPAELGALWREALARAQVSGDDVGHVVFGNVIQTEPRDMY LGRVAAVNGGVTINAPALTVNRLCGSGLQAIVSAAQTILLGDTDVAIGGGAESMSRAPYLAPAARWG ARMGDAGLVDMMLGALHDPFHRIHMGVTAENVAKEYDISRAQQDEAALESHRRASAAIKAGYFKDQI VPWSKGRKGDVTFDTDEHVRHDATIDDMTKLRPVFVKENGTVTAGNASGLNDAAAAVVMMERAEA ERRGLKPLARLVSYGHAGVDPKAMGIGPVPATKIALERAGLQVSDLDVIEANEAFAAQACAVTKALGL DPAKVNPNGSGISLGHPIGATGALITVKALHELNRVQGRYALVTMCIGGGQGIAAIFERIKLSGGGGS GGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKA IEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGD YQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQ GDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASDVMWEYKWENTGDAELYGPFTSA QMQTWVSEGYFPDGVYCRKLDPPGGQFYNSKRIDFDLYTGSAGSAAGSGEFGSAEAAAKEAAAKA

GSAGSAAGSGEFGSESDSVEFNNAISYVNKIKTRFLDHPEIYRSFLEILHTYQKEQLHTKGRPFRGMS EEEVFTEVANLFRGQEDLLSEFGQFLPEAKRGSG

7. Hexanoyl-CoA Synthetase (HCS) (SEQ ID NO:2Q9)

MGKNYKSLDSVVASDFIALGITSEVAETLHGRLAEIVCNYGAATPQTWINIANHILSPDLPFSLHQMLF YGCYKDFGPAPPAWIPDPEKVKSTNLGALLEKRGKEFLGVKYKDPISSFSHFQEFSVRNPEVYWRTV LMDEMKISFSKDPECILRRDDINNPGGSEWLPGGYLNSAKNCLNVNSNKKLNDTMIVWRDEGNDDL PLNKLTLDQLRKRVWLVGYALEEMGLEKGCAIAIDMPMHVDAVVIYLAIVLAGYVWSIADSFSAPEIS TRLRLSKAKAIFTQDHIIRGKKRIPLYSRVVEAKSPMAIVIPCSGSNIGAELRDGDISWDYFLERAKEFK NCEFTAREQPVDAYTNILFSSGTTGEPKAIPWTQATPLKAAADGWSHLDIRKGDVIVWPTNLGWMM GPWLVYASLLNGASIALYNGSPLVSGFAKFVQDAKVTMLGWPSIVRSWKSTNCVSGYDWSTIRCFS SSGEASNVDEYLWLMGRANYKPVIEMCGGTEIGGAFSAGSFLQAQSLSSFSSQCMGCTLYILDKNG YPMPKNKPGIGELALGPVMFGASKTLLNGNHHDVYFKGMPTLNGEVLRRHGDIFELTSNGYYHAHG RADDTMNIGGIKISSIEIERVCNEVDDRVFETTAIGVPPLGGGPEQLVIFFVLKDSNDTTIDLNQLRLSF NLGLQKKLNPLFKVTRVVPLSSLPRTATNKIMRRVLRQQFSHFEGSG

Figure 13B

GPP Gene Cassette Amino Acid Sequences

1. HMG-CoA Synthase (HMGS) - Enzyme Linker-cTPR6 Spacer-ID Linker - ID7 (SEQ ID NO:166)

MKLSTKLCWCGIKGRLRPQKQQQLHNTNLQMTELKKQKTAEQKTRPQNVGIKGIQIYIPTQCVNQSE LEKFDGVSQGKYTIGLGQTNMSFVNDREDIYSMSLTVLSKLIKSYNIDTNKIGRLEVGTETLIDKSKSVK SVLMQLFGENTDVEGIDTLNACYGGTNALFNSLNWIESNAWDGRDAIWCGDIAIYDKGAARPTGGA GTVAMWIGPDAPIVFDSVRASYMEHAYDFYKPDFTSEYPYVDGHFSLTCYVKALDQVYKSYSKKAIS KGLVSDPAGSDALNVLKYFDYNVFHVPTCKLVTKSYGRLLYNDFRANPQLFPEVDAELATRDYDESL TDKNIEKTFVNVAKPFHKERVAQSLIVPTNTGNMYTASVYAAFASLLNYVGSDDLQGKRVGLFSYGS GLAASLYSCKIVGDVQHIIKELDITNKLAKRITETPKDYEAAIELRENAHLKKNFKPQGSIEHLQSGVYYL TNIDDKFRRSYDVKKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDP NNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALEL DPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQ KALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGAS LGPLPPGWEVRSTVSGRIYFVDHNNRTTQFTDPRLHGSAGSAAGSGEFGSAEAAAKEAAAKAGSAG SAAGSGEFGSGAMGPLPPGWEKRTDSNGRVYFVNHNTRITQWEDPRSGSG

2. Truncated HMG-CoA Reductase (tHMGR) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID8 (SEQ

ID NO:167)

MVAVRRKALSILAEAPVLASDRLPYKNYDYDRVFGACCENVIGYMPLPVGVIGPLVIDGTSYHIPMATT EGCLVASAMRGCKAINAGGGATTVLTKDGMTRGPWRFPTLKRSGACKIWLDSEEGQNAIKKAFNS TSRFARLQHIQTCLAGDLLFMRFRTTTGDAMGMNMISKGVEYSLKQMVEEYGWEDMEWSVSGNY CTDKKPAAINWIEGRGKSWAEATIPGDWRKVLKSDVSALVELNIAKNLVGSAMAGSVGGFNAHAA NLVTAVFLALGQDPAQNVESSNCITLMKEVDGDLRISVSMPSIEVGTIGGGTVLEPQGAMLDLLGVRG PHATAPGTNARQLARIVACAVLAGELSLCAALAAGHLVQSHMTHNRKLSGGGGSGGGGSGGGGSA EAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPN NAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALE LDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQK ALELDPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSEFFRRERNKMAAAK CRNRRRELTDTLQAETDQLEDEKSALQTEIANLLKEKEKLEFILAAHRPACKIPDDLGFPEEMSLEGSA GSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVES LKQKIEELKQRKAQLKNDIANLEKEIAYAETGSG

3. Mevalonate Kinase (ERG12) - Enzyme Linker- cTPR6 Spacer- ID Linker- ID9 (SEQ ID NO:168)

MSLPFLTSAPGKVIIFGEHSAVYNKPAVAASVSALRTYLLISESSAPDTIELDFPDISFNHKWSINDFNAI TEDQVNSQKLAKAQQATDGLSQELVSLLDPLLAQLSESFHYHAAFCFLYMFVCLCPHAKNIKFSLKST LPIGAGLGSSASISVSLALAMAYLGGLIGSNDLEKLSENDKHIVNQWAFIGEKCIHGTPSGIDNAVATY GNALLFEKDSHNGTINTNNFKFLDDFPAIPMILTYTRIPRSTKDLVARVRVLVTEKFPEVMKPILDAMG ECALQGLEIMTKLSKCKGTDDEAVETNNELYEQLLELIRINHGLLVSIGVSHPGLELIKNLSDDLRIGST KLTGAGGGGCSLTLLRRDITQEQIDSFKKKLQDDFSYETFETDLGGTGCCLLSAKNLNKDLKIKSLVF QLFENKTTTKQQIDDLLLPGNTNLPWTSKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQ KAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGD YQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYY KQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGG SGGGGSGGGGASMEPAMEPETLEARINRATNPLNKELDWASINGFCEQLNEDFEGPPLATRLLAHKI QSPQEWEAIQALTVLETCMKSCGKRFHDEVGKFRFLNELIKWSPKYLGSRTSEKVKNKILELLYSWT VGLPEEVKIAEAYQMLKKQGIVKSGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSGA MGSMAEAEGESLESWLNKATNPSNRQEDWEYIIGFCDQINKELEGPQIAVRLLAHKIQSPQEWEALQ ALTVLEACMKNCGRRFHNEVGKFRFLNELIKWSPKYLGDRVSEKVKTKVIELLYSWTMALPEEAKIK DAYHMLKRQGIVQSDPPIPVDRTLIPSPPPRPKNGSG

4. Phosphomevalonate Kinase (ERG8) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID10 (SEQ ID NO:169)

MSELRAFSAPGKALLAGGYLVLDTKYEAFVVGLSARMHAVAHPYGSLQGSDKFEVRVKSKQFKDGE WLYHISPKSGFIPVSIGGSKNPFIEKVIANVFSYFKPNMDDYCNRNLFVIDIFSDDAYHSQEDSVTEHR GNRRLSFHSHRIEEVPKTGLGSSAGGLVTVLTTALASFFVSDLENNVDKYREVIHNLAQVAHCQAQG KIGSGFDVAAAAYGSIRYRRFPPALISNLPDIGSATYGSKLAHLVDEEDWNITIKSNHLPSGLTLWMGD IKNGSETVKLVQKVKNWYDSHMPESLKIYTELDHANSRFMDGLSKLDRLHETHDDYSDQIFESLERN DCTCQKYPEITEVRDAVATIRRSFRKITKESGADIEPPVQTSLLDDCQTLKGVLTCLIPGAGGYDAIAVI TKQDVDLRAQTANDKRFSKVQWLDVTQADWGVRKEKDPETYLDKKLSGGGGSGGGGSGGGGSA EAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPN NAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALE LDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQK ALELDPNNRSRSAGGGGSGGGGSGGGGASSYYHHHHHHLESTSLYKKAGSGSQKVEELKNKIAEL ENRNAVKKNRVAHLKQEIAYLKDELAAHEFEGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGS GEFGSSYYHHHHHHLESTSLYKKAGSGSFENVTHEFILATLENENAKLRRLEAKLERELARLRNEVA WLGSG

5. Diphosphomevalonate Decarboxylase (MVD1) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID11 (SEQ ID NQ:170)

MTVYTASVTAPVNIATLKYWGKRDTKLNLPTNSSISVTLSQDDLRTLTSAATAPEFERDTLWLNGEPH SIDNERTQNCLRDLRQLRKEMESKDASLPTLSQWKLHIVSENNFPTAAGLASSAAGFAALVSAIAKLY QLPQSTSEISRIARKGSGSACRSLFGGYVAWEMGKAEDGHDSMAVQIADSSDWPQMKACVLVVSDI KKDVSSTQGMQLTVATSELFKERIEHVVPKRFEVMRKAIVEKDFATFAKETMMDSNSFHATCLDSFP PIFYMNDTSKRIISWCHTINQFYGETIVAYTFDAGPNAVLYYLAENESKLFAFIYKLFGSVPGWDKKFT TEQLEAFNHQFESSNFTARELDLELQKDVARVILTQVGSGPQETNESLIDAKTGLPKEKLSGGGGSG GGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIE YYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDY QKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQ GDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASAMADLEQKVLEMEASTYDGVFIW KISDFPRKRQEAVAGRIPAIFSPAFYTSRYGYKMCLRIYLNGDGTGRGTHLSLFFWMKGPNDALLRW PFNQKVTLMLLDQNNREHVIDAFRPDVTSSSFQRPVNDMNIASGCPLFCPVSKMEAKNSYVRDDAIFI KAIVDLTGLGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSASIKLQSSDGEIFEVDVEI AKQSVTIKTMLEDLGMDDEGDDDPVPLPNVNAAILKKVIQWCTHHKDDPPPPEDDENKEKRTDDIPV WDQEFLKVDQGTLFELILAANYLDIKGLLDVTCKTVANMIKGKTPEEIRKTFNIKNDFTEEEEAQVRKE NQWCGSG

6. Isopentenvl-Diphosphate Delta-lsomerase (IDI1) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID12 (SEQ ID NQ:171)

MTADNNSMPHGAVSSYAKLVQNQTPEDILEEFPEIIPLQQRPNTRSSETSNDESGETCFSGHDEEQI KLMNENCIVLDWDDNAIGAGTKKVCHLMENIEKGLLHRAFSVFIFNEQGELLLQQRATEKITFPDLWT NTCCSHPLCIDDELGLKGKLDDKIKGAITAAVRKLDHELGIPEDETKTRGKFHFLNRIHYMAPSNEPW GEHEIDYILFYKINAKENLTVNPNVNEVRDFKWVSPNDLKTMFADPSYKFTPWFKIICENYLFNWWEQ LDDLSEVENDRQIHRMLKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALEL DPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKA LELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEY YQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGG ASSYYHHHHHHLESTSLYKKAGSGSNTVKELKNYIQELEERNAELKNLKEHLKFAKAELEFELAAHKF EGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQ KVAQLKNRVAYKLKENAKLENIVARLENDNANLEKDIANLEKDIANLERDVARGSG

7. Geranvl-Diphosphate Synthase (ERG20ww) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID13 (SEQ ID NO:172)

MEAKIDELINNDPVWSSQNESLISKPYNHILLKPGKNFRLNLIVQINRVMNLPKDQLAIVSQIVELLHNS SLLIDDIEDNAPLRRGQTTSHLIWGVPSTINTANYMYFRAMQLVSQLTTKEPLYHWLITIFNEELINLHR GQGLDIYWRDFLPEIIPTQEMYLNMVMNKTGGLFRLTLRLMEALSPSSHHGHSLVPFINLLGIIYQIRD DYLNLKDFQMSSEKGFAEDITEGKLSFPIVHALNFTKTKGQTEQHNEILRILLLRTSDKDIKLKLIQILEF DTNSLAYTKNFINQLVNMIKNDNENKYLPDLASHSDTATNLHDELLYIIDHLSELKLSGGGGSGGGGS GGGGSAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQK ALELDPNNAEAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIE YYQKALELDPNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQK AIEDYQKALELDPNNRSRSAGGGGSGGGGSGGGGASLCTMKKGPSGYGFNLHSDKSKPGQFIRSV DPDSPAEASGLRAQDRIVEVNGVCMEGKQHGDWSAIRAGGDETKLLWDREGSAGSAAGSGEFG SAEAAAKEAAAKAGSAGSAAGSGEFGSSSGAIIYTVELKRYGGPLGITISGTEEPFDPIIISSLTKGGLA ERTGAIHIGDRILAINSSSLKGKPLSEAIHLLQMAGETVTLKIKKQTDAQPASSGSG

Figure 13C

CAN Gene Cassette Amino Acid Sequences

1. Olivetol Synthase (OS) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID14 (SEQ ID NO:175)

MNHLRAEGPASVLAIGTANPENILLQDEFPDYYFRVTKSEHMTQLKEKFRKICDKSMIRKRNCFLNEE HLKQNPRLVEHEMQTLDARQDMLVVEVPKLGKDACAKAIKEWGQPKSKITHLIFTSASTTDMPGADY HCAKLLGLSPSVKRVMMYQLGCYGGGTVLRIAKDIAENNKGARVLAVCCDIMACLFRGPSESDLELL VGQAIFGDGAAAVIVGAEPDESVGERPIFELVSTGQTILPNSEGTIGGHIREAGLIFDLHKDVPMLISNN IEKCLIEAFTPIGISDWNSIFWITHPGGKAILDKVEEKLHLKSDKFVDSRHVLSEHGNMSSSTVLFVMDE LRKRSLEEGKSTTGDGFEWGVLFGFGPGLTVERVWRSVPIKYKLSGGGGSGGGGSGGGGSAEA WYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNA EAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELD PNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKAL ELDPNNRSRSAGGGGSGGGGSGGGGASGNNLETYEWYNKSISRDKAEKLLLDTGKEGAFMVRDS RTPGTYTVSVFTKAIISENPCIKHYHIKETNDSPKRYYVAEKYVFDSIPLLIQYHQYNGGGLVTRLRYPV CGGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSGSHPWFFGKIPRAKAEEMLSKQR HDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLVWVKFNSLNELVDYHRSTSVSRNQ QIFLRDIEQVPQQPTGSG

2. Olivetolic Acid Cyclase (OAC) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID15 (SEQ ID NO: 176)

MAVKHLIVLKFKDEITEAQKEEFFKTYVNLVNIIPAMKDVYWGKDVTQKNKEEGYTHIVEVTFESVETI QDYIIHPAHVGFGDVYRSFWEKLLIFDYTPRKKLSGGGGSGGGGSGGGGSAEAWYNLGNAYYKQG DYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYK QGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNASAWYNLGN AYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDPNNRSRSAG GGGSGGGGSGGGGASGQDRSEATLIKRFKGEGVRYKAKLIGIDEVSAARGDKLCQDSMMKLKGVV AGARSKGEHKQKIFLTISFGGIKIFDEKTGALQHHHAVHEISYIAKDITDHRAFGYVCGKEGNHRFVAIK TAQAAEPVILDLRDLFQLIYELKQREELEKKAGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGS GEFGSGSHMGSQFVWTSQKTEASERCGLQGSYILRVEAEKLTLLTLGAQSQILEPLLFWPYTLLRRY GRDKVMFSFEAGRRCPSGPGTFTFQTSQGNDIFQAVEAAIQQQKAQGKVGQAQDILRLEHHHHHH GSG

3. CBGA Synthase (CBGAS) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID16 (SEQ ID NO:177)

MGLSSVCTFSFQTNYHTLLNPHNNNPKTSLLCYRHPKTPIKYSYNNFPSKHCSTKSFHLQNKCSESL SIAKNSIRAATTNQTEPPESDNHSVATKILNFGKACWKLQRPYTIIAFTSCACGLFGKELLHNTNLISW SLMFKAFFFLVAILCIASFTTTINQIYDLHIDRINKPDLPLASGEISVNTAWIMSnVALFGLimKIViKGGPL YIFGYCFGFGGIVYSVPPFRWKQNPSTAFLLNFLAHHTNFTFYYASRAALGLRFELRPSFTFLLAFMK SMGSALALIKDASDVEGDTKFGISTLASKYGSRNLTLFCSGIVLLSYVAAILAGIIWPQAFNSNVMLLSH AILAFWULQTRDFALTNYDPEAGRRFYEFMWKLYYAEYLVYVFIKLSGGGGSGGGGSGGGGSAEA WYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNA EAWYNLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELD PNNASAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKAL ELDPNNRSRSAGGGGSGGGGSGGGGASAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQWW NAEDSEGKRGMIPVPYVEKYGSAGSAAGSGEFGSAEAAAKEAAAKAGSAGSAAGSGEFGSLIKHMR AEALFDFTGNSKLELNFKAGDVIFLLSRINKDWLEGTVRGATGIFPLSFVKILKGSG

4. CBDA Synthase (CBDAS) (SEQ ID NO:173)

MKCSTFSFWFVCKIIFFFFSFNIQTSIANPRENFLKCFSQYIPNNATNLKLVYTQNNPLYMSVLNSTIHN LRFTSDTTPKPLVIVTPSHVSHIQGTILCSKKVGLQIRTRSGGHDSEGMSYISQVPFVIVDLRNMRSIKI DVHSQTAWVEAGATLGEVYYWVNEKNENLSLAAGYCPTVCAGGHFGGGGYGPLMRNYGLAADNII

DAHLVNVHGKVLDRKSMGEDLFWALRGGGAESFGIIVAWKIRLVAVPKSTMFSVKKIMEIHELVKLVN KWQNIAYKYDKDLLLMTHFITRNITDNQGKNKTAIHTYFSSVFLGGVDSLVDLMNKSFPELGIKKTDCR QLSWIDTIIFYSGWNYDTDNFNKEILLDRSAGQNGAFKIKLDYVKKPIPESVFVQILEKLYEEDIGAGM YALYPYGGIMDEISESAIPFPHRAGILYELWYICSWEKQEDNEKHLNWIRNIYNFMTPYVSKNPRLAYL NYRDLDIGINDPKNPNNYTQARIWGEKYFGKNFDRLVKVKTLVDPNNFFRNEQSIPPLPRHRHGSG

5. CBCA Synthase (CBCAS) (SEQ ID NO:174)

MNCSTFSFWFVCKIIFFFLSFNiQ!SIANPQENFLKCFSEYIPNNPANPKFIYTQHDQLYMSVLNS&amp;#1470;nQNL RFTSDTTPKPLWTPSNVSHIQASILCSKKVGLQIRTRSGGHDAEGLSYISQVPFAIVDLRNMHTVKV DIHSQTAVWEAGATLGEVYYWiNEMNENFSFPGGYCPTVGVGGHFSGGGYGALMRNYGLAADNIlD AHLVNVDGKVLDRKSMGEDLFWAIRGGGGENFGIIAACKIKLWVPSKATIFSVKKNMEIHGLVKLFNK WQNIAYKYDKDLMLTTHFRTRNITDNHGKNKTTVHGYFSSIFLGGVDSLVDLMNKSFPELGIKKTDCK ELSWIDTTIFYSGWNYNTANFKKEILLDRSAGKKTAFSIKLDYVKKLIPETAMVKILEKLYEEEVGVGM YVLYPYGGIMDEISESAIPFPHRAGIMYELWYTATWEKQEDNEKHINVWRSVYNFTTPYVSQNPRLA YLNYRDLDLGKTNPESPNNYTQARIWGEKYFGKNFNRLVKVKTKADPNNFFRNEQSIPPLPPRHHGS G

6. Acetyl-CoA Carboxylase (ACC) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID17 (SEQ ID

NO:178)

MSEESLFESSPQKMEYEITNYSERHTELPGHFIGLNTVDKLEESPLRDFVKSHGGHTVISKILIANNGIA AVKEIRSVRKWAYETFGDDRTVQFVAMATPEDLEANAEYIRMADQYIEVPGGTNNNNYANVDLIVDIA ERADVDAVWAGWGHASENPLLPEKLSQSKRKVIFIGPPGNAMRSLGDKISSTIVAQSAKVPCIPWSG TGVDTVHVDEKTGLVSVDDDIYQKGCCTSPEDGLQKAKRIGFPVMIKASEGGGGKGIRQVEREEDFI ALYHQAANEIPGSPIFIMKLAGRARHLEVQLLADQYGTNISLFGRDCSVQRRHQKIIEEAPVTIAKAETF HEMEKAAVRLGKLVGYVSAGTVEYLYSHDDGKFYFLELNPRLQVEHPTTEMVSGVNLPAAQLQIAM GIPMHRISDIRTLYGMNPHSASEIDFEFKTQDATKKQRRPIPKGHCTACRITSEDPNDGFKPSGGTLH ELNFRSSSNVWGYFSVGNNGNIHSFSDSQFGHIFAFGENRQASRKHMVVALKELSIRGDFRTTVEYL IKLLETEDFEDNTITTGWLDDLITHKMTAEKPDPTLAVICGAATKAFLASEEARHKYIESLQKGQVLSKD LLQTMFPVDFIHEGKRYKFTVAKSGNDRYTLFINGSKCDIILRQLSDGGLLIAIGGKSHTIYWKEEVAAT RLSVDSMTTLLEVENDPTQLRTPSPGKLVKFLVENGEHIIKGQPYAEIEVMKMQMPLVSQENGIVQLL KQPGSTIVAGDIMAIMTLDDPSKVKHALPFEGMLPDFGSPVIEGTKPAYKFKSLVSTLENILKGYDNQV IMNASLQQLIEVLRNPKLPYSEWKLHISALHSRLPAKLDEQMEELVARSLRRGAVFPARQLSKLIDMA VKNPEYNPDKLLGAVVEPLADIAHKYSNGLEAHEHSIFVHFLEEYYEVEKLFNGPNVREENIILKLRDE NPKDLDKVALTVLSHSKVSAKNNLILAILKHYQPLCKLSSKVSAIFSTPLQHIVELESKATAKVALQAREI LIQGALPSVKERTEQIEHILKSSWKVAYGSSNPKRSEPDLNILKDLIDSNYVVFDVLLQFLTHQDPWT AAAAQVYIRRAYRAYTIGDIRVHEGVTVPIVEWKFQLPSAAFSTFPTVKSKMGMNRAVSVSDLSYVAN SQSSPLREGILMAVDHLDDVDEILSQSLEVIPRHQSSSNGPAPDRSGSSASLSNVANVCVASTEGFE SEEEILVRLREILDLNKQELINASIRRITFMFGFKDGSYPKYYTFNGPNYNENETIRHIEPALAFQLELGR LSNFNIKPIFTDNRNIHVYEAVSKTSPLDKRFFTRGIIRTGHIRDDISIQEYLTSEANRLMSDILDNLEVTD TSNSDLNHIFINFIAVFDISPEDVEAAFGGFLERFGKRLLRLRVSSAEIRIIIKDPQTGAPVPLRALINNVS GYVIKTEMYTEVKNAKGEVWFKSLGKPGSMHLRPIATPYPVKEWLQPKRYKAHLMGTTYVYDFPELF RQASSSQWKNFSADVKLTDDFFISNELIEDENGELTEVEREPGANAIGMVAFKITVKTPEYPRGRQFV WANDITFKIGSFGPQEDEFFNKVTEYARKRGIPRIYLAANSGARIGMAEEIVPLFQVAWNDAANPDK GFQYLYLTSEGMETLKKFDKENSVLTERTVINGEERFVIKTIIGSEDGLGVECLRGSGLIAGATSRAYH DIFTITLVTCRSVGIGAYLVRLGQRAIQVEGQPIILTGAPAINKMLGREVYTSNLQLGGTQIMYNNGVSH LTAVDDLAGVEKIVEWMSYVPAKRNMPVPILETKDTWDRPVDFTPTNDETYDVRWMIEGRETESGF EYGLFDKGSFFETLSGWAKGVWGRARLGGIPLGVIGVETRTVENLIPADPANPNSAETLIQEPGQV WHPNSAFKTAQAINDFNNGEQLPMMILANWRGFSGGQRDMFNEVLKYGSFIVDALVDYKQPIIIYIPP TGELRGGSVWVVDPTINADQMEMYADVNARAGVLEPQGMVGIKFRREKLLDTMNRLDDKYRELRS QLSNKSLAPEVHQQISKQLADRERELLPIYGQISLQFADLHDRSSRMVAKGVISKELEWTEARRFFFW RLRRRLNEEYLIKRLSHQVGEASRLEKIARIRSWYPASVDHEDDRQVATWIEENYKTLDDKLKGLKLE

SFAQDLAKKIRSDHDNAIDGLSEVIKMLSTDDKEKLLKTLKKLSGGGGSGGGGSGGGGSAEAWYNL GNAYYKQGDYQKAIEYYQKALELDPNNAEAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAEAWY NLGNAYYKQGDYQKAIEDYQKALELDPNNLQAEAWKNLGNAYYKQGDYQKAIEYYQKALELDPNNA SAWYNLGNAYYKQGDYQKAIEYYQKALELDPNNAKAWYRRGNAYYKQGDYQKAIEDYQKALELDP NNRSRSAGGGGSGGGGSGGGGASGSHMRLGAQSIQPTANLDRTDDLVYLNVMELVRAVLELKNEL AQLPPEGYWVVKNVGLTLRKLIGSVDDLLPSLPSSSRTEIEGTQKLLNKDLAELINKMRLAQQNAVTS LSEECKRQMLTASHTLAVDAKNLLDAVDQAKVLANLAHPPAEGSAGSAAGSGEFGSAEAAAKEAAA KAGSAGSAAGSGEFGSGAMATPGSENVLPREPLIATAVKFLQNSRVRQSPLATRRAFLKKKGLTDEE IDMAFQQSGTAADEPSSLWGSG

Figure 13D

SCF Gene Cassette Amino Acid Sequences

1. Cannabinoidergic Metabolon Scaffold (CBSCFLD) - (Myc)3(SEQ ID NO:179)

MGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSARNA YLRKKIARLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASHEQGSAGSAAGSGEFAEAAAKEAAA KAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENETLKKKNLHKKD LIAYLEKEIANLRKKIEEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGS SYYHHHHHHLESTSLYKKAGSGSQKVAELKNRVAVKLNRNEQLKNKVEELKNRNAYLKNELATLENE VARLENDVAEGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKA GSGSNEVTTLENDAAFIENENAYLEKEIARLRKEKAALRNRLAHKKGSAGSAAGSGEFGSAEAAAKE AAAKEAAAKEAAAKAGSAGSAAGSGEFGSRPPTISNPPPLISSAKHPSVGSAGSAAGSGEFAEAAAK EAAAKAGSAGSAAGSGEFNFLQSRPEPTAPPEESFRSGGSAGSAAGSGEFGSAEAAAKEAAAKEA AAKEAAAKAGSAGSAAGSGEFGSSKGTGLNPNAKVWQEIAPGNGSAGSAAGSGEFAEAAAKEAAA KAGSAGSAAGSGEFPDGGTTFEHLWSSLEPDSTYGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKE AAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSKRIAYLRKKIAALKKDNANLEKDIA NLENEIERLIKEIKTLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHH HHHLESTSLYKKAGSGSNLLATLRSTAAVLENENHVLEKEKEKLRKEKEQLLNKLEAYKGSAGSAAG SGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSPATSQHPPPPPGHRSQAPSHGSA GSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFELNSLLILLEAAEYLERRDRGSAGSAAGSGEFG SAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSRPPTISNPPPLISSAKHPSVGSAGSAAGSG EFAEAAAKEAAAKAGSAGSAAGSGEFNFLQSRPEPTAPPEESFRSGGSAGSAAGSGEFGSAEAAAK EAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSKGTGLNPNAKVWQEIAPGNGSAGSAAGSGEFAEA AAKEAAAKAGSAGSAAGSGEFPDGGTTFEHLWSSLEPDSTYGSAGSAAGSGEFGSAEAAAKEAAA KEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSKRIAYLRKKIAALKKDNA NLEKDIANLENEIERLIKEIKTLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEF SYYHHHHHHLESTSLYKKAGSGSNLLATLRSTAAVLENENHVLEKEKEKLRKEKEQLLNKLEAYKGS AGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSALVDDAADYEPPPSNNE EALGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFRELFDDPSYVNVQNLDKARQGSAGSA AGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSKNTKSMNFDNPVYRKTTEEEG SAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFRSLPSTWIENKLYGMSDPNWGSAGSAAGSG EFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSVVDNSPPPALPPKKRQSAPSGSAGSA AGSGEFAEAAAKEAAAKAGSAGSAAGSGEFTQRSKPQPAVPPRPSADLILGSAGSAAGSGEFGSAE AAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSTDEEREETEEEVYLLNSTTLGSAGSAAGSGEF AEAAAKEAAAKAGSAGSAAGSGEFDGNVSGTQRLDSATVRTYSCGSAGSAAGSGEFGSAEAAAKE AAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGSGSQKVAQLKNRVAYKL KENAKLENIVARLENDNANLEKDIANLEKDIANLERDVARGSAGSAAGSGEFAEAAAKEAAAKAGSA GSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNTVKELKNYIQELEERNAELKNLKEHLKFAKAELE FELAAHKFEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSHDDSLPH PQQATDDSGHESDGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFGSPNAGSVEQTPKKP GLRRRGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHL ESTSLYKKAGSGSFENVTHEFILATLENENAKLRRLEAKLERELARLRNEVAWLGSAGSAAGSGEFA EAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSQKVEELKNKIAELENRNAVK KNRVAHLKQEIAYLKDELAAHEFEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSA AGSGEFGSVSSTKLVSFHDDSDEDLLHIGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFAA ATPISTFHDDSDEDLLHVGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEF GSSYYHHHHHHLESTSLYKKAGSGSQKVESLKQKIEELKQRKAQLKNDIANLEKEIAYAETGSAGSAA GSGEFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSEFFRRERNKMAAAKC RNRRRELTDTLQAETDQLEDEKSALQTEIANLLKEKEKLEFILAAHRPACKIPDDLGFPEEMSLEGSA

GSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSFQMPADTPPPAYLPPEDP MTGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFERESNEEPPPPYEDPYWGNGGSAGSA AGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSSYYHHHHHHLESTSLYKKAGS GSQKVAELKNRVAVKLNRNEQLKNKVEELKNRNAYLKNELATLENEVARLENDVAEGSAGSAAGSG EFAEAAAKEAAAKAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNEVTTLENDAAFIENENA YLEKEIARLRKEKAALRNRLAHKKSYYHHHHHHLESTSLYKKAGSGSARNAYLRKKIARLKKDNLQLE RDEQNLEKIIANLRDEIARLENEVASHEQGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFSY YHHHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENETLKKKNLHKKDLIAYLEKEIANLRKKIEEG SAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSEQKLISEEDLEQKLISEE DLEQKLISEEDLGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEFGSG

2. Malonyl-CoA Metabolon Scaffold (MCASCFLD) - (FLAGMSEQ ID NQ:180)

MGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSARNA YLRKKIARLKKDNLQLERDEQNLEKIIANLRDEIARLENEVASHEQGSAGSAAGSGEFAEAAAKEAAA KAGSAGSAAGSGEFSYYHHHHHHLESTSLYKKAGSGSNLVAQLENEVASLENENETLKKKNLHKKD LIAYLEKEIANLRKKIEEGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGS SATRELDELMASLSDFKIQGGSAGSAAGSGEFAEAAAKEAAAKAGSAGSAAGSGEFDLALSENWAQ EFLAAGDAVDGSAGSAAGSGEFGSAEAAAKEAAAKEAAAKEAAAKAGSAGSAAGSGEFGSDYKDD DDKDYKDDDDKDYKDDDDKGSAGSAAGSGEFGSAGSAAGSGEFGSAGSAAGSGEFGSG

Figure 14A

Codon-optimized HCA Gene Cassette Nucleotide Sequences

1. ATP Citrate Lyase (ACL) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID1 (SEQ ID NQ:181)

ATGAGTGCTAAGGCAATTTCTGAACAAACTGGTAAAGAATTGTTGTACAAGTTTATTTGTACTACA TCAGCCATCCAAAATAGATTCAAATACGCTAGAGTTACCCCAGATACTGACTGGGCTAGATTGTT ACAAGATCATCCATGGTTGTTATCTCAAAACTTGGTTGTCAAACCTGACCAATTAATTAAGAGAAG AGGTAAATTGGGTTTAGTAGGTGTTAATTTGACATTGGATGGTGTAAAGTCTTGGTTGAAACCAA GATTAGGTCAAGAAGCCACAGTTGGTAAAGCTACCGGTTTCTTGAAAAATTTCTTGATCGAACCA TTTGTCCCTCATTCACAAGCCGAAGAATTCTATGTATGTATCTACGCTACTAGAGAGGGTGACTA TGTTTTATTTCATCACGAAGGTGGTGTCGACGTAGGTGACGTTGACGCCAAGGCTCAAAAGTTGT TGGTTGGTGTCGATGAAAAGTTGAACCCAGAAGACATTAAAAAGCATTTGTTGGTTCACGCACCT GAAGATAAAAAGGAAATATTGGCCTCCTTTATAAGTGGTTTGTTTAATTTCTACGAAGATTTGTAC TTCACCTACTTGGAAATTAACCCATTAGTAGTTACTAAGGATGGTGTATATGTTTTGGACTTAGCT GCAAAAGTTGATGCAACAGCCGACTACATTTGTAAGGTCAAATGGGGTGACATCGAATTTCCACC TCCATTCGGTAGAGAAGCTTATCCAGAAGAAGCCTACATTGCTGATTTGGACGCTAAGTCTGGTG CATCATTGAAGTTGACATTGTTGAACCCTAAAGGTAGAATTTGGACCATGGTTGCTGGTGGTGGT GCTAGTGTCGTATATTCTGATACTATATGCGACTTGGGTGGTGTTAACGAATTGGCAAACTACGG TGAATACTCAGGTGCCCCATCCGAACAACAAACATACGATTACGCTAAGACCATCTTGTCCTTAA TGACTAGAGAAAAGCATCCTGATGGTAAAATCTTGATCATCGGTGGTAGTATCGCAAATTTTACT AACGTTGCCGCTACATTCAAGGGTATCGTCAGAGCTATAAGAGATTACCAAGGTCCATTGAAGG AACACGAAGTAACAATATTCGTTAGAAGAGGTGGTCCTAACTACCAAGAAGGTTTGAGAGTCATG GGTGAAGTAGGTAAAACCACTGGTATACCAATCCATGTCTTTGGTACAGAAACCCACATGACTGC AATAGTTGGTATGGCCTTAGGTCATAGACCAATCCCTAATCAACCTCCAACCGCAGCCCACACTG CAAATTTCTTGTTAAACGCCTCTGGTTCAACTTCCACACCAGCTCCTTCTAGAACAGCAAGTTTCT CTGAATCAAGAGCTGATGAAGTCGCTCCAGCTAAGAAAGCAAAACCAGCCATGCCTCAAGACTC CGTTCCAAGTCCTAGATCTTTGCAGGGTAAATCTACTACTTTGTTTTCTAGACATACTAAGGCTAT AGTATGGGGTATGCAAACAAGAGCAGTTCAAGGCATGTTGGATTTCGACTATGTTTGTAGTAGAG ATGAACCATCTGTTGCTGCAATGGTCTATCCTTTTACTGGTGACCATAAGCAAAAATTCTACTGG GGTCACAAGGAAATATTGATCCCAGTTTTTAAGAACATGGCCGATGCTATGAGAAAACATCCTGA AGTCGACGTATTGATTAACTTCGCCTCATTAAGATCCGCTTACGATTCTACAATGGAAACCATGA ACTACGCTCAAATAAGAACCATCGCTATCATTGCAGAAGGTATTCCAGAAGCCTTGACTAGAAAG TTGATTAAGAAAGCTGATCAAAAAGGTGTCACAATAATCGGTCCAGCTACCGTAGGTGGTATTAA GCCTGGTTGTTTCAAGATCGGTAACACTGGTGGTATGTTGGATAACATATTGGCATCTAAGTTGT ATAGACCAGGTTCAGTCGCTTACGTATCCAGAAGTGGTGGTATGTCCAACGAATTGAACAACATC ATCAGTAGAACTACAGATGGTGTATACGAAGGTGTTGCTATTGGTGGTGACAGATACCCAGGTT CTACTTTTATGGATCATGTATTGAGATATCAAGACACACCTGGTGTTAAAATGATTGTTGTCTTGG GTGAAATAGGTGGTACTGAAGAATACAAGATATGCAGAGGTATCAAAGAAGGTAGATTGACAAA GCCAATCGTTTGTTGGTGCATTGGTACTTGTGCAACAATGTTTTCTTCAGAAGTTCAATTCGGTCA TGCAGGTGCCTGCGCTAATCAAGCTTCAGAAACAGCAGTTGCCAAGAACCAAGCATTAAAAGAA GCCGGTGTTTTTGTCCCTAGATCTTTCGATGAATTAGGTGAAATCATTCAATCAGTCTATGAAGAC TTGGTAGCTAATGGTGTAATTGTTCCAGCACAAGAAGTTCCTCCACCTACTGTCCCTATGGATTA CTCTTGGGCAAGAGAATTGGGTTTAATTAGAAAGCCAGCTAGTTTTATGACCTCTATATGTGATG AAAGAGGTCAAGAATTGATCTATGCTGGTATGCCTATTACTGAAGTATTCAAAGAAGAAATGGGT ATCGGTGGTGTTTTAGGTTTGTTGTGGTTCCAAAAGAGATTGCCAAAGTACTCTTGTCAATTCATT GAAATGTGCTTAATGGTTACAGCTGATCATGGTCCTGCTGTCTCAGGTGCACACAATACCATAAT CTGCGCTAGAGCTGGTAAAGATTTGGTTTCTTCTTTGACCTCAGGTTTGTTAACTATTGGTGACA GATTTGGTGGTGCATTAGACGCCGCTGCAAAGATGTTTTCAAAAGCTTTCGATTCCGGTATAATC CCAATGGAATTCGTTAATAAGATGAAAAAGGAGGGTAAATTGATAATGGGTATCGGTCATCGTGT TAAGTCTATCAATAACCCTGATATGAGAGTACAAATCTTGAAGGACTATGTTAGACAACACTTTCC AGCCACACCTTTGTTAGATTACGCTTTGGAAGTTGAAAAGATTACCACTTCTAAAAAGCCAAATTT

GATCTTGAACGTTGATGGTTTAATTGGTGTTGCTTTTGTCGACATGTTGAGAAACTGTGGTTCCTT CACTAGAGAAGAAGCTGATGAATATATCGACATTGGTGCATTGAATGGTATCTTTGTTTTAGGTA GATCTATGGGTTTCATTGGTCATTACTTGGATCAAAAGAGATTAAAGCAAGGTTTGTACAGACAT CCATGGGATGACATTTCTTACGTTTTACCTGAACACATGTCAATGAAATTGTCTGGTGGTGGTGG TTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCTTGGTACAATTTGGGTAACGCA TACTACAAGCAGGGTGACTACCAAAAGGCAATTGAATATTACCAAAAGGCCTTGGAATTAGACCC AAATAACGCAGAAGCCTGGTATAATTTGGGTAATGCTTATTATAAACAGGGTGACTATCAAAAGG CTATCGAATACTACCAAAAGGCATTGGAATTAGACCCTAATAACGCTGAAGCATGGTATAATTTG GGTAACGCTTATTATAAGCAGGGTGACTATCAAAAAGCCATCGAAGACTACCAAAAGGCTTTGGA ATTAGATCCAAATAACTTACAAGCCGAAGCTTGGAAGAATTTGGGTAACGCTTACTATAAACAGG GTGACTACCAAAAAGCAATTGAATACTATCAAAAAGCTTTAGAATTGGACCCTAATAACGCATCA GCCTGGTACAATTTGGGTAATGCTTACTATAAGCAGGGTGACTATCAGAAGGCCATTGAATACTA TCAAAAGGCTTTAGAATTGGATCCAAATAACGCTAAAGCATGGTACAGACGTGGTAACGCTTATT ACAAACAGGGTGACTACCAGAAAGCCATTGAAGATTATCAAAAGGCTTTGGAATTGGATCCTAAC AACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTT CTTCATATTACCATCACCATCACCATCACTTGGAATCCACAAGTTTATACAAAAAGGCTGGTTCTG GTTCAAATTTGGTCGCACAATTGGAAAACGAAGTAGCCTCTTTAGAAAATGAAAACGAAACCTTG AAAAAGAAAAACTTACATAAGAAAGATTTGATCGCTTATTTGGAAAAGGAAATCGCAAATTTGAGA AAGAAAATTGAAGAAGGTAGTGCAGGTTCTGCCGCTGGTTCTGGTGAATTTGGTTCAGCTGAAG CAGCCGCTAAGGAAGCAGCCGCTAAAGCCGGTTCAGCTGGTTCCGCAGCCGGTTCTGGTGAAT TCGGTTCCAGTTACTATCACCATCACCATCATCACTTGGAATCCACTAGTTTATATAAGAAAGCAG GTTCTGGTTCAGCAAGAAATGCCTACTTGAGAAAGAAAATAGCTAGATTAAAGAAAGATAACTTG CAATTGGAAAGAGATGAACAAAATTTGGAAAAGATTATCGCCAACTTAAGAGATGAAATCGCTAG ATTGGAAAATGAAGTTGCATCCCATGAACAAGGTAGTGGT

2. Acetyl-CoA Acetyltransferase (atoB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID2 (SEQ ID NO:182)

ATGAAAAACTGTGTAATCGTTTCTGCTGTTAGAACTGCAATTGGTTCCTTTAATGGTAGTTTGGCC TCTACATCAGCTATTGATTTGGGTGCTACCGTCATCAAAGCTGCAATTGAAAGAGCAAAGATTGA TTCTCAACATGTCGACGAAGTAATAATGGGTAACGTTTTGCAAGCTGGTTTAGGTCAAAATCCAG CAAGACAAGCCTTGTTAAAATCTGGTTTAGCAGAAACTGTATGTGGTTTCACAGTTAATAAGGTCT GCGGTTCTGGTTTGAAGTCAGTTGCTTTAGCCGCTCAAGCTATACAAGCAGGTCAAGCCCAATCT ATCGTCGCTGGTGGTATGGAAAATATGTCATTGGCACCTTATTTGTTAGATGCAAAAGCCAGATC AGGTTATAGATTAGGTGACGGTCAAGTATACGACGTTATTTTGAGAGATGGTTTAATGTGCGCTA CTCATGGTTATCACATGGGTATTACAGCAGAAAATGTTGCCAAAGAATACGGTATAACCAGAGAA ATGCAAGATGAATTGGCATTACATTCCCAAAGAAAGGCAGCCGCTGCAATCGAAAGTGGTGCTT TTACTGCAGAAATTGTCCCAGTAAACGTTGTCACAAGAAAGAAAACTTTCGTTTTCTCCCAAGATG AATTCCCAAAAGCTAATAGTACCGCTGAAGCATTGGGTGCTTTAAGACCTGCATTCGACAAGGCC GGTACCGTAACTGCCGGTAATGCTTCTGGTATAAACGATGGTGCCGCTGCATTGGTTATCATGG AAGAATCAGCCGCTTTAGCAGCCGGTTTGACACCTTTAGCTAGAATTAAATCTTATGCATCAGGT GGTGTTCCACCTGCTTTGATGGGTATGGGTCCAGTCCCTGCTACCCAAAAGGCATTGCAATTAG CCGGTTTGCAATTGGCTGATATCGACTTAATCGAAGCAAACGAAGCCTTTGCTGCACAATTCTTG GCAGTTGGTAAAAATTTGGGTTTCGACTCCGAAAAGGTTAATGTCAACGGTGGTGCCATTGCTTT GGGTCATCCAATAGGTGCTTCAGGTGCAAGAATCTTGGTTACATTGTTGCATGCCATGCAAGCTA GAGATAAAACCTTGGGTTTAGCTACTTTGTGTATCGGTGGTGGTCAAGGTATCGCAATGGTTATC GAAAGATTGAATAAGTTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTA GTGCAGAAGCCTGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAAAAGGCAAT CGAATACTACCAAAAGGCCTTGGAATTAGATCCAAATAACGCTGAAGCATGGTATAATTTGGGTA ATGCCTATTATAAACAGGGTGACTATCAAAAAGCTATTGAATATTACCAAAAGGCATTGGAATTAG ATCCTAATAACGCCGAAGCTTGGTATAATTTGGGTAACGCCTATTATAAGCAGGGTGACTATCAA AAGGCCATCGAAGATTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTGCAAGCAGAAGCCT

GGAAGAATTTGGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTATTGAATACTATCAA AAAGCCTTAGAATTGGATCCTAATAACGCTTCTGCATGGTACAATTTGGGTAATGCCTACTATAAA CAGGGTGACTACCAGAAGGCTATTGAATACTACCAAAAAGCATTAGAATTGGATCCAAATAACGC CAAGGCTTGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTACCAGAAAGCCATAGAA GACTATCAAAAAGCCTTGGAATTGGATCCTAACAACAGATCCAGAAGTGCTGGTGGTGGTGGTT CTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCATATTACCATCACCATCACCATCACTTG GAATCTACATCATTATACAAAAAGGCTGGTTCCGGTAGTAATGAAGTTACTACATTGGAAAACGA TGCCGCTTTTATCGAAAACGAAAACGCATACTTGGAAAAGGAAATCGCCAGATTAAGAAAGGAAA AGGCAGCCTTGAGAAATAGATTAGCCCATAAAAAGGGTTCCGCTGGTAGTGCTGCAGGTTCTGG TGAATTTGGTTCAGCTGAAGCCGCTGCAAAAGAAGCCGCTGCAAAGGCAGGTTCTGCCGGTTCA GCCGCTGGTTCTGGTGAATTCGGTTCCAGTTACTATCACCATCACCATCATCACTTGGAATCTAC TTCATTATATAAAAAGGCCGGTTCCGGTAGTCAAAAAGTCGCTGAATTAAAGAACAGAGTAGCTG TTAAGTTGAACAGAAACGAACAATTGAAAAATAAGGTAGAAGAATTGAAAAATAGAAACGCCTAC TTAAAGAATGAATTGGCAACATTGGAAAACGAAGTCGCTAGATTGGAAAATGATGTAGCAGAAGG TTCTGGT

3. 3-Hvdroxvbutvrvl-CoA Dehydrogenase (BHBD) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID3 (SEQ ID NO:183)

ATGAAAAAGGTTTGTGTCATTGGTGCTGGTACCATGGGTTCTGGTATAGCACAAGCCTTTGCTGC AAAAGGTTTCGAAGTTGTCTTGAGAGATATCAAGGACGAATTCGTTGATAGAGGTTTGGACTTCA TCAATAAGAACTTGTCTAAGTTGGTTAAAAAGGGTAAAATCGAAGAAGCTACAAAGGTAGAAATC TTGACCAGAATTTCAGGTACTGTTGATTTGAATATGGCCGCTGATTGTGACTTGGTAATCGAAGC AGCCGTTGAAAGAATGGATATTAAGAAACAAATCTTCGCAGATTTGGACAACATCTGCAAACCTG AAACAATCTTAGCCTCAAACACCTCTTCATTGTCCATTACTGAAGTCGCTAGTGCAACAAAAAGA CCAGATAAGGTAATAGGCATGCATTTCTTTAATCCAGCTCCTGTTATGAAGTTGGTAGAAGTTATT AGAGGTATAGCAACATCTCAAGAAACCTTTGACGCTGTTAAGGAAACTTCAATAGCAATCGGTAA AGATCCAGTCGAAGTAGCCGAAGCTCCTGGTTTCGTAGTTAACAGAATCTTGATACCTATGATCA ACGAAGCTGTTGGTATCTTGGCTGAAGGTATTGCATCTGTCGAAGATATTGACAAAGCCATGAAG TTAGGTGCTAATCACCCAATGGGTCCTTTGGAATTGGGTGACTTTATTGGTTTGGACATATGTTTA GCTATCATGGACGTTTTGTATTCCGAAACAGGTGACAGTAAATACAGACCACATACCTTGTTGAA GAAATATGTTAGAGCAGGTTGGTTAGGTAGAAAGTCTGGTAAAGGTTTCTACGATTACTCTAAAA AGTTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCT GGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAAAAGGCCATAGAATACTACCAA AAGGCTTTGGAATTGGATCCTAATAACGCTGAAGCATGGTATAATTTGGGTAATGCATATTATAAA CAGGGTGACTATCAAAAGGCAATCGAATACTACCAAAAGGCCTTGGAATTAGATCCAAATAACGC CGAAGCTTGGTATAATTTGGGTAACGCCTATTATAAGCAGGGTGACTATCAAAAAGCTATCGAAG ATTACCAAAAGGCATTGGAATTGGATCCTAACAACTTACAAGCAGAAGCCTGGAAGAATTTGGGT AACGCATATTACAAACAGGGTGACTACCAAAAAGCCATTGAATATTATCAAAAAGCTTTGGAATTG GATCCAAATAACGCTTCAGCATGGTACAATTTGGGTAATGCCTATTACAAGCAGGGTGACTATCA GAAAGCTATTGAATATTATCAAAAGGCTTTGGAATTAGATCCTAATAACGCCAAGGCTTGGTACA GACGTGGTAATGCCTATTACAAGCAGGGTGACTACCAGAAGGCCATTGAAGACTATCAAAAAGC CTTGGAATTGGATCCAAACAACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGT TCTGGTGGTGGTGGTGCTTCCGAAAATTTGTACTTCCAAGGTGAAAACTTGTACTTCCAGGGTGA CTCCAGTGAAAGTTGTTGGAATTGCGGTAGAAAAGCCTCCGAAACCTGTAGTGGTTGCAACACT GCTAGATATTGTGGTTCTTTTTGCCAACACAAAGATTGGGAAAAGCATCACCATATTTGTGGTCA AACATTACAAGCACAACAAGGTTCTGCCGGTTCAGCTGCAGGTTCTGGTGAATTTGGTTCCGCT GAAGCCGCTGCAAAAGAAGCCGCTGCAAAGGCAGGTTCCGCCGGTAGTGCCGCTGGTAGTGGT GAATTCGGTTCTATGGCAGTTTCCGAAAGTCAATTGAAGAAAATGGTTTCTAAGTACAAGTACAG AGATTTGACTGTTAGAGAAACAGTTAACGTCATCACTTTGTACAAGGATTTGAAGCCAGTCTTGG ACTCATACGTTTTTAATGATGGTTCTTCAAGAGAATTGATGAACTTAACTGGTACAATACCAGTTC CTTACCGTGGTAACACTTACAACATCCCAATCTGTTTGTGGTTGTTAGATACATATCCTTACAATC

CACCTATCTGCTTCGTCAAACCAACATCCAGTATGACCATTAAAACTGGTAAACATGTTGATGCTA

ACGGTAAAATATATTTGCCATACTTACACGAATGGAAGCATCCTCAATCAGACTTGTTGGGTTTAA

TCCAAGTAATGATCGTCGTATTTGGTGACGAACCACCTGTTTTCTCTAGACCAGGTTCAGGT

4. Enoyl-CoA Hydratase (ECH) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID4 (SEQ ID NO:184)

ATGGAATTGAACAACGTTATATTGGAAAAGGAGGGTAAAGTCGCTGTTGTCACTATAAATAGACC AAAGGCATTGAACGCCTTGAACTCTGATACATTGAAGGAAATGGACTACGTTATCGGTGAAATTG AAAACGATTCAGAAGTCTTAGCAGTAATTTTGACCGGTGCCGGTGAAAAATCCTTTGTTGCCGGT GCTGATATCAGTGAAATGAAGGAAATGAACACTATCGAAGGTAGAAAGTTCGGTATCTTGGGTAA CAAGGTTTTCAGAAGATTGGAATTGTTGGAAAAGCCTGTTATAGCTGCAGTCAATGGTTTCGCTT TGGGTGGTGGTTGTGAAATCGCAATGTCCTGCGATATTAGAATAGCTTCTTCAAACGCAAGATTT GGTCAACCAGAAGTCGGTTTAGGTATTACACCTGGTTTCGGTGGTACCCAAAGATTATCTAGATT GGTTGGTATGGGTATGGCCAAGCAATTGATTTTTACTGCTCAAAACATCAAGGCTGATGAAGCAT TGAGAATCGGTTTGGTTAATAAGGTAGTTGAACCATCTGAATTGATGAACACCGCCAAGGAAATC GCTAATAAGATTGTTTCTAATGCTCCAGTTGCTGTCAAGTTGAGTAAGCAAGCTATAAATCGTGG TATGCAATGTGATATCGACACTGCATTGGCCTTCGAATCTGAAGCATTTGGTGAATGCTTCTCAA CAGAAGATCAAAAAGACGCAATGACCGCCTTTATCGAAAAGAGAAAGATAGAAGGTTTCAAAAAC AGAAAGTTATCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAG CATGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAAAAGGCAATCGAATACTAC CAAAAGGCCTTGGAATTGGACCCAAATAACGCCGAAGCTTGGTATAATTTGGGTAATGCCTATTA TAAACAGGGTGACTATCAAAAAGCTATAGAATACTACCAAAAGGCATTGGAATTGGACCCTAATA ACGCAGAAGCCTGGTATAATTTGGGTAACGCCTATTATAAGCAGGGTGACTATCAAAAGGCCATA GAAGACTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTACAAGCTGAAGCATGGAAGAATTT GGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTATTGAATATTATCAAAAAGCTTTAGA ATTAGACCCTAATAACGCCTCTGCTTGGTACAATTTGGGTAATGCCTACTATAAACAGGGTGACT ACCAGAAGGCTATTGAATATTACCAAAAAGCTTTAGAATTGGATCCAAATAACGCAAAGGCCTGG TACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTACCAGAAAGCCATTGAAGATTATCAAAA AGCTTTGGAATTGGATCCTAACAACAGATCCAGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGT GGTTCTGGTGGTGGTGGTGCTTCTGGTCCATTGGGTTCCCCTTTGACTGCATCAATGTTAGCTTC CGCACCACCTCAAGAACAAAAGCAAATGTTGGGTGAAAGATTATTCCCATTGATACAAGCTATGC ATCCTACTTTAGCAGGTAAAATCACAGGCATGTTGTTGGAAATCGATAACTCTGAATTGTTACACA TGTTAGAATCCCCAGAAAGTTTGAGATCTAAAGTTGACGAAGCCGTAGCTGTTTTGCAAGCTCAT CAAGCAAAAGAAGCCGCTCAAAAGGCCGGTTCAGCTGGTTCCGCAGCCGGTAGTGGTGAATTT GGTTCTGCTGAAGCTGCAGCCAAAGAAGCTGCAGCCAAGGCAGGTAGTGCCGGTTCTGCTGCA GGTTCTGGTGAATTCGGTTCCAATACCAACATGAGTGTCCCAACTGATGGTGCTGTAACTACATC TCAAATTCCTGCATCAGAACAAGAAACTTTAGTTAGACCAAAGCCTTTGTTGTTGAAGTTGTTGAA GTCAGTAGGTGCTCAAAAAGATACCTACACTATGAAGGAAGTTTTATTTTATTTGGGTCAATACAT CATGACAAAGAGATTATACGATGAAAAGCAACAACATATCGTTTACTGTTCAAACGATTTGTTGG GTGACTTGTTTGGTGTACCATCTTTCTCAGTTAAGGAACACAGAAAGATCTATACAATGATATACA GAAATTTGGTCGTAGGTTCTGGT

5. Trans-Enoyl-CoA Reductase (ECR) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID5 (SEQ ID

NO:185)

ATGATCGTAAAGCCAATGGTTAGAAACAACATCTGTTTGAACGCTCATCCTCAAGGTTGCAAAAA GGGTGTAGAAGATCAAATCGAATACACCAAAAAGAGAATCACTGCAGAAGTTAAAGCCGGTGCT AAAGCACCTAAGAATGTTTTGGTCTTAGGTTGTTCCAACGGTTATGGTTTGGCTAGTAGAATAAC AGCTGCATTTGGTTACGGTGCCGCTACCATCGGTGTTTCCTTCGAAAAGGCTGGTAGTGAAACC AAATATGGTACTCCAGGTTGGTACAATAACTTGGCATTTGATGAAGCAGCCAAGAGAGAAGGTTT ATACTCTGTCACTATAGATGGTGACGCTTTCTCAGATGAAATCAAGGCACAAGTTATTGAAGAAG CCAAAAAGAAAGGTATAAAATTCGATTTGATCGTTTACTCCTTAGCAAGTCCAGTCAGAACAGAT CCTGACACCGGTATAATGCATAAGTCTGTTTTGAAGCCATTCGGTAAAACTTTCACAGGTAAAAC

AGTCGATCCTTTCACCGGTGAATTGAAAGAAATATCTGCTGAACCAGCAAATGATGAAGAAGCTG CAGCCACAGTAAAAGTTATGGGTGGTGAAGACTGGGAAAGATGGATCAAGCAATTGTCCAAAGA AGGTTTGTTGGAAGAAGGTTGTATCACCTTAGCTTATTCATACATTGGTCCTGAAGCCACTCAAG CTTTGTATAGAAAAGGTACAATCGGTAAAGCTAAAGAACATTTGGAAGCCACCGCTCACAGATTA AATAAGGAAAACCCATCTATCAGAGCATTTGTTTCTGTAAATAAGGGTTTAGTTACTAGAGCATCC GCCGTTATCCCAGTCATTCCTTTGTATTTGGCTAGTTTGTTTAAGGTTATGAAGGAAAAGGGTAAC CATGAAGGTTGCATAGAACAAATCACTAGATTGTACGCAGAAAGATTATACAGAAAGGATGGTAC AATTCCAGTTGACGAAGAAAACAGAATCAGAATCGATGACTGGGAATTGGAAGAAGATGTCCAAA AGGCAGTATCTGCCTTAATGGAAAAAGTTACCGGTGAAAACGCTGAATCATTGACTGATTTGGCA GGTTATAGACACGACTTTTTAGCCTCTAATGGTTTCGATGTCGAAGGTATTAACTACGAAGCAGA AGTAGAAAGATTCGACAGAATTAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGT GGTGGTGGTAGTGCTGAAGCATGGTATAATTTGGGTAACGCTTATTACAAGCAGGGTGACTACC AAAAGGCCATCGAATACTACCAAAAGGCTTTGGAATTGGACCCTAATAACGCCGAAGCTTGGTA CAATTTGGGTAATGCCTACTATAAACAGGGTGACTATCAAAAAGCAATTGAATATTACCAAAAGG CCTTGGAATTAGACCCAAATAACGCAGAAGCCTGGTACAATTTGGGTAACGCCTACTATAAGCAG GGTGACTATCAAAAGGCTATTGAAGACTACCAAAAGGCATTGGAATTAGATCCTAATAACTTGCA AGCTGAAGCATGGAAAAATTTGGGTAATGCCTATTATAAACAGGGTGACTACCAAAAAGCTATTG AATACTATCAAAAAGCTTTGGAATTGGACCCAAATAACGCCTCAGCTTGGTATAATTTGGGTAAT GCATACTACAAACAGGGTGACTATCAGAAGGCAATAGAATACTATCAAAAAGCCTTAGAATTGGA TCCTAATAACGCAAAAGCCTGGTATAGACGTGGTAATGCCTACTACAAGCAGGGTGACTATCAG AAGGCGATAGAAGATTATCAAAAGGCATTGGAATTGGATCCAAACAACAGATCTAGATCAGCTGG TGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCATATTACCATCACCAT CACCATCACTTGGAATCCACAAGTTTATATAAGAAAGCAGGTTCTGGTTCAAATTTGTTAGCCACT TTGAGATCAACAGCTGCAGTATTGGAAAACGAAAACCATGTTTTGGAAAAAGAAAAGGAAAAGTT GAGAAAGGAAAAGGAACAATTGTTGAATAAGTTGGAAGCCTACAAAGGTTCTGCTGGTTCAGCC GCTGGTTCCGGTGAATTCGGTAGTGCTGAAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCTGGT TCCGCAGGTAGTGCAGCCGGTTCTGGTGAATTTGGTTCCAGTTACTATCACCATCACCATCATCA CTTGGAATCCACTAGTTTATATAAGAAAGCTGGTTCTGGTTCAAAGAGAATCGCATACTTGAGAA AGAAAATCGCTGCATTAAAGAAAGATAACGCCAACTTGGAAAAGGACATCGCTAATTTGGAAAAC GAAATCGAAAGATTGATTAAAGAAATTAAAACATTAGAAAATGAAGTTGCTTCTCATGAACAAGGT TCAGGT

6. Beta-Ketothiolase (bktB) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID6 (SEQ ID NO: 186)

ATGACTAGAGAAGTTGTCGTAGTTAGTGGTGTTAGAACAGCTATTGGTACCTTTGGTGGTTCTTT AAAAGATGTTGCACCAGCCGAATTGGGTGCATTAGTCGTAAGAGAAGCTTTGGCAAGAGCCCAA GTTTCAGGTGACGATGTCGGTCATGTTGTCTTCGGTAACGTTATCCAAACAGAACCAAGAGATAT GTATTTGGGTAGAGTAGCTGCAGTTAATGGTGGTGTTACCATAAACGCTCCTGCATTAACTGTCA ACAGATTGTGTGGTAGTGGTTTACAAGCTATTGTTTCTGCCGCTCAAACAATATTGTTAGGTGAC ACCGACGTTGCTATCGGTGGTGGTGCTGAATCTATGTCAAGAGCCCCATACTTAGCTCCTGCAG CCAGATGGGGTGCCAGAATGGGTGACGCTGGTTTGGTTGACATGATGTTGGGTGCTTTGCATGA TCCATTCCATAGAATCCACATGGGTGTAACTGCAGAAAACGTTGCCAAGGAATACGATATCTCAA GAGCACAACAAGACGAAGCTGCATTAGAATCACACAGAAGAGCATCCGCCGCTATTAAAGCCGG TTACTTTAAGGATCAAATAGTTCCAGTAGTTTCTAAAGGTAGAAAGGGTGACGTTACCTTCGATAC TGACGAACATGTTAGACACGACGCTACTATTGATGACATGACAAAGTTAAGACCTGTTTTCGTCA AGGAAAATGGTACTGTTACAGCTGGTAATGCATCTGGTTTGAACGATGCAGCCGCTGCAGTCGT AATGATGGAAAGAGCCGAAGCTGAAAGAAGAGGTTTGAAACCATTAGCTAGATTGGTTTCTTATG GTCATGCTGGTGTCGATCCTAAAGCAATGGGTATAGGTCCAGTTCCTGCTACTAAGATCGCATTG GAAAGAGCCGGTTTACAAGTCTCTGATTTGGACGTAATTGAAGCCAATGAAGCTTTTGCCGCTCA AGCATGTGCCGTTACAAAAGCCTTGGGTTTAGATCCAGCTAAGGTCAATCCTAACGGTAGTGGT ATCTCTTTAGGTCATCCAATTGGTGCAACCGGTGCCTTGATAACTGTTAAGGCTTTGCACGAATT GAACAGAGTACAAGGTAGATATGCATTAGTTACAATGTGCATCGGTGGTGGTCAAGGTATTGCA

GCCATATTCGAAAGAATTAAGTTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTG GTGGTAGTGCTGAAGCATGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAAAA GGCAATCGAATATTACCAAAAAGCCTTGGAATTAGACCCAAATAACGCCGAAGCTTGGTATAATT TGGGTAATGCCTATTATAAACAGGGTGACTATCAAAAAGCTATAGAATACTACCAAAAGGCATTG GAATTAGACCCTAATAACGCAGAAGCCTGGTATAATTTGGGTAACGCCTATTATAAGCAGGGTGA CTATCAAAAGGCCATAGAAGACTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTACAAGCTG AAGCATGGAAGAATTTGGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTATTGAATAC TATCAAAAGGCTTTAGAATTGGACCCTAATAACGCCTCTGCTTGGTACAATTTGGGTAATGCCTA CTATAAACAGGGTGACTACCAGAAGGCTATCGAATATTATCAAAAAGCTTTAGAATTGGACCCAA ATAACGCAAAGGCCTGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTACCAGAAAGC TATTGAAGATTATCAAAAGGCATTGGAATTGGATCCTAACAACAGATCCAGAAGTGCTGGTGGTG GTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGATGTTATGTGGGAATATAAGTG GGAAAATACAGGTGACGCTGAATTATACGGTCCTTTTACTTCAGCACAAATGCAAACATGGGTAT CCGAAGGTTATTTCCCTGATGGTGTTTACTGCAGAAAATTAGACCCACCTGGTGGTCAATTCTAC AACTCAAAGAGAATAGATTTCGACTTGTACACCGGTTCAGCTGGTTCCGCTGCAGGTTCTGGTG AATTTGGTTCCGCAGAAGCCGCTGCAAAAGAAGCCGCTGCAAAGGCTGGTAGTGCAGGTTCTG CCGCTGGTAGTGGTGAATTTGGTTCTGAATCAGATTCCGTCGAATTCAATAACGCTATATCTTAC GTAAATAAGATTAAAACCAGATTTTTAGATCATCCAGAAATCTATAGATCATTCTTAGAAATCTTGC ATACATACCAAAAAGAACAATTGCACACCAAGGGTAGACCTTTCAGAGGCATGTCCGAAGAAGA AGTCTTTACTGAAGTAGCTAATTTGTTTAGAGGTCAAGAAGATTTGTTGTCAGAATTCGGTCAATT CTTGCCAGAAGCAAAAAGAGGTTCCGGT

7. Hexanoyl-CoA Synthetase (HCS) (SEQ ID NO:187)

ATGGGTAAAAATTACAAGTCATTGGATTCCGTTGTCGCAAGTGACTTTATTGCCTTGGGTATAACT TCTGAAGTCGCAGAAACATTGCATGGTAGATTAGCCGAAATTGTATGTAACTACGGTGCTGCAAC CCCACAAACTTGGATCAACATAGCAAACCATATCTTGTCACCAGATTTGCCTTTCTCCTTGCACC AAATGTTGTTTTATGGTTGCTACAAGGATTTCGGTCCTGCTCCACCTGCATGGATTCCAGACCCT GAAAAGGTTAAGTCAACTAATTTGGGTGCTTTGTTAGAAAAGAGAGGTAAAGAATTCTTGGGTGT TAAGTACAAGGATCCAATCTCTTCTTTTTCTCACTTCCAAGAATTTTCTGTCAGAAACCCTGAAGT ATACTGGAGAACAGTTTTGATGGATGAAATGAAAATAAGTTTCTCTAAGGACCCAGAATGTATCTT GAGAAGAGATGACATCAACAACCCAGGTGGTTCTGAATGGTTGCCAGGTGGTTATTTGAACTCA GCTAAAAATTGCTTGAACGTTAACTCCAATAAGAAATTGAATGATACTATGATTGTCTGGAGAGAT GAAGGCAACGATGACTTGCCATTGAATAAGTTGACATTGGATCAATTGAGAAAGAGAGTTTGGTT GGTCGGTTACGCATTAGAAGAAATGGGTTTGGAAAAAGGTTGTGCCATAGCTATCGATATGCCTA TGCATGTAGACGCTGTAGTTATCTATTTGGCTATTGTTTTAGCAGGTTACGTCGTAGTTTCTATAG CTGATTCATTTTCCGCACCAGAAATCTCAACTAGATTGAGATTATCCAAAGCAAAGGCCATATTCA CACAAGATCACATCATCAGAGGTAAAAAGAGAATCCCTTTATACTCAAGAGTCGTAGAAGCCAAA TCCCCAATGGCTATAGTTATCCCTTGTAGTGGTTCTAACATTGGTGCAGAATTAAGAGATGGTGA CATATCTTGGGATTACTTTTTGGAAAGAGCCAAAGAATTCAAGAATTGCGAATTCACTGCCAGAG AACAACCAGTTGATGCTTACACTAACATTTTGTTCTCCAGTGGTACTACAGGTGAACCAAAAGCA ATACCTTGGACACAAGCCACCCCTTTAAAGGCCGCTGCAGATGGTTGGTCACATTTGGATATTAG AAAAGGTGACGTCATAGTATGGCCAACTAATTTGGGTTGGATGATGGGTCCTTGGTTGGTTTATG CTAGTTTGTTAAATGGTGCCTCTATTGCTTTATACAACGGTAGTCCATTGGTTTCTGGTTTCGCTA AATTTGTCCAAGATGCAAAAGTAACAATGTTGGGTGTTGTCCCTTCAATCGTTAGAAGTTGGAAG TCTACAAATTGTGTCTCAGGTTATGATTGGTCCACCATCAGATGCTTTTCTTCATCCGGTGAAGC CTCTAATGTCGACGAATATTTGTGGTTAATGGGTAGAGCTAACTACAAGCCAGTTATCGAAATGT GTGGTGGTACCGAAATTGGTGGTGCATTCTCAGCCGGTTCCTTTTTACAAGCTCAATCATTGAGT TCTTTTTCATCCCAATGTATGGGTTGCACATTGTACATCTTGGATAAGAACGGTTACCCAATGCCT AAAAATAAGCCAGGTATTGGTGAATTGGCTTTAGGTCCTGTTATGTTCGGTGCATCTAAAACATT GTTGAACGGTAACCATCACGATGTATACTTCAAGGGTATGCCAACCTTAAATGGTGAAGTTTTGA GAAGACATGGTGACATATTCGAATTAACCTCAAACGGTTACTACCATGCCCACGGTAGAGCTGAT

GACACTATGAACATCGGTGGTATCAAAATCAGTTCTATCGAAATCGAAAGAGTATGTAACGAAGT TGATGACAGAGTCTTTGAAACCACTGCAATTGGTGTTCCACCATTGGGTGGTGGTCCAGAACAAT TAGTAATCTTTTTCGTTTTGAAGGATTCTAACGACACAACCATAGATTTGAACCAATTGAGATTAT CTTTTAACTTGGGTTTACAAAAGAAATTGAACCCATTATTCAAAGTTACTAGAGTAGTTCCATTGT CATCCTTACCTAGAACTGCTACAAACAAGATTATGAGAAGAGTCTTGAGACAACAATTCAGTCATT TTGAAGGTTCTGGT

Figure 14B Codon-optimized GPP Gene Cassette Nucleotide Sequences

1. HMG-CoA Synthase (HMGS) - Enzyme Linker-cTPR6 Spacer-ID Linker - ID7 (SEQ ID NO:188)

ATGAAGTTATCTACTAAATTGTGTTGGTGCGGTATTAAGGGTAGATTAAGACCACAAAAGCAACA ACAATTGCATAACACAAACTTGCAAATGACCGAATTGAAGAAACAAAAGACTGCTGAACAAAAGA CTAGACCACAAAACGTTGGTATTAAAGGTATCCAAATCTATATCCCTACACAATGTGTCAATCAAT CTGAATTGGAAAAGTTTGATGGTGTATCACAGGGTAAATACACTATCGGTTTAGGTCAAACAAAC ATGTCTTTCGTAAACGATAGAGAAGACATCTATTCTATGTCATTGACTGTTTTGTCCAAGTTGATA AAAAGTTACAACATCGATACAAACAAGATTGGTAGATTGGAAGTTGGTACCGAAACTTTGATCGA TAAGTCCAAGAGTGTCAAGTCTGTATTGATGCAATTGTTCGGTGAAAATACCGATGTTGAAGGTA TCGACACTTTAAATGCTTGTTATGGTGGTACTAACGCATTATTCAATTCATTGAACTGGATCGAAT CCAATGCCTGGGATGGTAGAGATGCTATTGTTGTCTGCGGTGACATCGCTATCTATGACAAAGG TGCTGCAAGACCAACCGGTGGTGCAGGTACTGTTGCCATGTGGATAGGTCCAGATGCACCTATC GTTTTTGACTCTGTCAGAGCATCATACATGGAACATGCCTACGATTTCTACAAACCAGACTTCAC CTCCGAATATCCTTACGTTGATGGTCACTTTTCTTTGACTTGTTACGTCAAGGCTTTGGACCAAGT ATACAAGTCTTACTCTAAGAAAGCAATATCTAAGGGTTTGGTTTCAGATCCAGCTGGTTCCGACG CATTAAACGTCTTGAAGTACTTCGATTACAACGTTTTCCATGTCCCTACATGCAAGTTGGTTACCA AGTCTTACGGTAGATTGTTGTACAACGATTTCAGAGCTAACCCACAATTGTTCCCTGAAGTCGAC GCTGAATTAGCAACTAGAGATTACGACGAATCTTTGACAGATAAGAACATCGAAAAGACTTTCGT AAACGTTGCAAAGCCATTCCACAAAGAAAGAGTTGCCCAATCATTAATTGTCCCTACAAATACCG GTAACATGTATACAGCCTCAGTTTACGCCGCTTTTGCTTCCTTGTTAAATTATGTAGGTAGTGATG ACTTGCAAGGTAAAAGAGTTGGTTTATTCTCCTATGGTAGTGGTTTAGCAGCCTCTTTGTACTCTT GTAAGATTGTAGGTGACGTTCAACACATTATTAAGGAATTGGACATCACTAATAAGTTGGCTAAG AGAATCACTGAAACACCAAAGGATTATGAAGCTGCAATCGAATTGAGAGAAAACGCACATTTGAA GAAAAATTTCAAACCTCAAGGTAGTATAGAACACTTGCAATCTGGTGTCTACTACTTAACAAACAT CGATGACAAATTCAGAAGATCATACGATGTTAAAAAGAAATTGTCTGGTGGTGGTGGTTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTATAATTTGGGTAACGCTTATTACAA GCAGGGTGACTACCAAAAAGCAATCGAATATTACCAAAAGGCCTTGGAATTAGACCCAAATAACG CCGAAGCTTGGTACAATTTGGGTAATGCATACTATAAACAGGGTGACTATCAAAAGGCTATCGAA TACTACCAAAAGGCATTGGAATTAGACCCTAATAACGCAGAAGCCTGGTACAATTTGGGTAACGC CTACTATAAGCAGGGTGACTATCAAAAAGCCATAGAAGACTACCAAAAGGCTTTGGAATTAGATC CAAATAACTTGCAAGCTGAAGCATGGAAAAATTTGGGTAATGCCTACTACAAACAGGGTGACTAC CAAAAGGCAATTGAATATTATCAAAAAGCCTTGGAATTAGATCCTAATAACGCCTCAGCTTGGTAT AATTTGGGTAATGCCTATTATAAGCAGGGTGACTACCAGAAAGCCATTGAATATTATCAAAAGGC TTTAGAATTGGATCCAAATAACGCAAAAGCCTGGTATAGACGTGGTAATGCCTACTACAAGCAGG GTGACTATCAGAAGGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAACAACAGATCC AGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTTGGGT CCTTTGCCACCTGGTTGGGAAGTAAGATCCACAGTTAGTGGTAGAATCTATTTCGTTGATCATAA CAACAGAACTACACAATTCACCGACCCAAGATTGCACGGTTCTGCTGGTTCAGCCGCTGGTTCT GGTGAATTTGGTTCCGCAGAAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCCGGTTCCGCTGGT AGTGCAGCCGGTAGTGGTGAATTTGGTTCTGGTGCTATGGGTCCATTACCACCTGGTTGGGAAA AGAGAACAGATTCTAACGGTAGAGTCTACTTCGTAAACCATAATACCAGAATTACTCAATGGGAA GATCCTAGATCTGGTTCAGGT

2. Truncated HMG-CoA Reductase (tHMGR) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID8 (SEQ

ID NO:189)

ATGGTAGCCGTTAGAAGAAAGGCTTTGTCTATCTTAGCCGAAGCTCCAGTTTTGGCATCAGATAG ATTACCTTACAAGAACTACGATTACGACAGAGTATTTGGTGCTTGTTGCGAAAACGTTATTGGTTA TATGCCATTGCCTGTCGGTGTAATCGGTCCATTAGTTATTGATGGTACATCTTACCATATCCCTAT GGCAACTACAGAAGGTTGTTTGGTTGCATCAGCCATGAGAGGTTGCAAGGCAATTAATGCTGGT GGTGGTGCTACCACTGTTTTAACCAAAGATGGTATGACTAGAGGTCCAGTTGTCAGATTTCCTAC TTTGAAGAGATCCGGTGCTTGTAAAATATGGTTAGATAGTGAAGAAGGTCAAAATGCCATCAAAA AGGCTTTTAACTCCACTAGTAGATTCGCAAGATTGCAACATATTCAAACATGCTTAGCCGGTGAC TTGTTGTTTATGAGATTCAGAACAACCACTGGTGACGCTATGGGTATGAATATGATATCTAAGGG TGTCGAATACTCATTGAAGCAAATGGTAGAAGAATACGGTTGGGAAGATATGGAAGTAGTTTCTG TTTCAGGCAACTACTGTACTGACAAAAAGCCAGCTGCAATTAACTGGATAGAAGGTCGTGGTAAA TCTGTCGTAGCTGAAGCAACAATACCTGGTGACGTTGTTAGAAAGGTTTTGAAATCTGACGTATC AGCTTTGGTTGAATTGAACATCGCTAAAAATTTGGTTGGTTCCGCCATGGCTGGTAGTGTCGGTG GTTTTAATGCACATGCCGCTAACTTAGTTACAGCAGTCTTCTTGGCCTTAGGTCAAGATCCAGCT CAAAACGTAGAATCTTCAAACTGTATCACCTTGATGAAAGAAGTTGATGGTGACTTAAGAATATCC GTTAGTATGCCATCAATAGAAGTCGGTACAATCGGTGGTGGTACCGTCTTGGAACCTCAAGGTG CAATGTTAGATTTGTTAGGTGTTAGAGGTCCACATGCAACTGCCCCTGGTACAAATGCTAGACAA TTGGCAAGAATTGTCGCTTGTGCAGTATTAGCTGGTGAATTGTCCTTATGCGCAGCCTTGGCTGC AGGTCACTTAGTTCAAAGTCATATGACACACAACAGAAAGTTGTCTGGTGGTGGTGGTTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCTTGGTATAATTTGGGTAACGCATATTACAA GCAGGGTGACTACCAAAAGGCCATCGAATACTACCAAAAGGCTTTGGAATTGGACCCAAATAAC GCAGAAGCCTGGTACAATTTGGGTAATGCTTACTATAAACAGGGTGACTATCAAAAGGCAATTGA ATATTACCAAAAGGCCTTGGAATTAGACCCTAATAACGCTGAAGCATGGTACAATTTGGGTAACG CCTACTATAAGCAGGGTGACTATCAAAAAGCTATTGAAGACTACCAAAAGGCATTGGAATTAGAT CCAAATAACTTGCAAGCCGAAGCTTGGAAAAATTTGGGTAACGCTTACTACAAACAGGGTGACTA CCAAAAAGCTATTGAATACTATCAAAAAGCTTTGGAATTGGACCCTAATAACGCATCTGCCTGGT ATAATTTGGGTAATGCTTATTATAAACAGGGTGACTACCAGAAGGCAATAGAATACTATCAAAAAG CCTTGGAATTAGACCCAAATAACGCTAAAGCATGGTATAGACGTGGTAATGCTTACTATAAGCAG GGTGACTACCAGAAAGCTATAGAAGATTATCAAAAGGCATTGGAATTGGATCCTAACAACAGATC TAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCCAGTTAT TACCATCACCATCACCATCACTTGGAATCCACTAGTTTATACAAAAAGGCAGGTTCAGAATTTTTC AGAAGAGAAAGAAATAAGATGGCCGCTGCAAAATGTAGAAACAGAAGAAGAGAATTGACAGATA CCTTACAAGCTGAAACCGATCAATTGGAAGACGAAAAGTCTGCATTGCAAACTGAAATAGCCAAT TTGTTGAAGGAAAAGGAAAAGTTGGAATTCATTTTAGCCGCTCATAGACCAGCTTGCAAAATTCC TGATGACTTGGGTTTCCCAGAAGAAATGTCTTTAGAAGGTTCCGCAGGTAGTGCAGCCGGTTCC GGTGAATTTGGTAGTGCTGAAGCTGCAGCCAAGGAAGCTGCAGCCAAAGCTGGTTCTGCAGGTT CAGCTGCAGGTTCCGGTGAATTCGGTTCTTCATACTATCACCATCACCATCATCACTTGGAATCT ACCTCATTATACAAAAAGGCTGGTTCCGGTAGTCAAAAGGTTGAATCTTTGAAGCAAAAGATTGA AGAATTGAAGCAAAGAAAAGCCCAATTGAAGAATGATATCGCTAACTTAGAAAAGGAAATCGCCT ACGCTGAAACTGGTTCTGGT

3. Mevalonate Kinase (ERG12) - Enzyme Linker- cTPR6 Spacer- ID Linker- ID9 (SEQ ID NQ:190)

ATGAGTTTACCATTTTTGACATCTGCTCCTGGTAAAGTTATTATATTCGGTGAACATAGTGCCGTC TATAATAAGCCAGCTGTCGCTGCATCTGTATCAGCTTTGAGAACATACTTGTTGATCTCTGAATCT TCAGCACCTGATACCATCGAATTGGATTTCCCAGACATCTCATTCAATCACAAGTGGTCCATTAAT GATTTCAACGCTATCACCGAAGACCAAGTAAACTCACAAAAGTTGGCCAAAGCTCAACAAGCAAC TGATGGTTTGTCACAAGAATTAGTTTCCTTGTTAGACCCATTGTTGGCTCAATTGTCCGAAAGTTT CCATTACCACGCCGCTTTCTGTTTCTTGTACATGTTCGTTTGTTTATGCCCTCATGCTAAGAATAT CAAATTTTCTTTGAAGTCTACTTTGCCAATTGGTGCAGGTTTAGGTTCCAGTGCCTCTATATCAGT

TTCCTTAGCATTGGCCATGGCTTATTTGGGTGGTTTGATAGGTAGTAACGATTTGGAAAAGTTGT CTGAAAACGACAAGCATATCGTCAACCAATGGGCATTCATCGGTGAAAAATGCATTCACGGTACT CCTAGTGGTATAGATAATGCAGTTGCCACATATGGTAACGCTTTGTTATTCGAAAAGGACTCTCA TAACGGTACCATCAACACTAACAACTTCAAGTTCTTGGATGACTTTCCTGCAATACCAATGATCTT GACTTACACAAGAATTCCAAGATCTACTAAAGATTTGGTAGCTAGAGTCAGAGTATTGGTTACAG AAAAGTTCCCTGAAGTTATGAAGCCAATCTTGGATGCAATGGGTGAATGTGCCTTGCAAGGTTTG GAAATCATGACAAAGTTGTCAAAGTGCAAGGGTACTGATGACGAAGCTGTTGAAACAAATAACGA ATTGTACGAACAATTGTTGGAATTGATCAGAATCAATCATGGTTTGTTAGTTTCAATTGGTGTCTC CCACCCAGGTTTAGAATTGATAAAGAACTTGTCAGATGACTTAAGAATCGGTTCCACAAAATTGA CCGGTGCTGGTGGTGGTGGTTGTTCTTTGACCTTGTTAAGAAGAGATATCACTCAAGAACAAATC GACAGTTTTAAAAAGAAATTGCAAGATGACTTCTCTTACGAAACTTTCGAAACAGATTTGGGTGGT ACTGGTTGTTGCTTGTTGTCAGCTAAGAATTTGAACAAAGATTTGAAGATTAAATCCTTGGTTTTC CAATTGTTCGAAAATAAGACTACAACCAAGCAACAAATCGATGACTTGTTGTTGCCTGGTAATAC AAACTTGCCATGGACCTCAAAATTATCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGT GGTGGTAGTGCTGAAGCATGGTATAATTTGGGTAACGCATATTACAAGCAGGGTGACTACCAAA AGGCTATCGAATACTACCAAAAGGCATTGGAATTGGACCCTAATAACGCCGAAGCTTGGTACAAT TTGGGTAATGCTTACTATAAACAGGGTGACTATCAAAAGGCCATTGAATATTACCAAAAGGCTTT GGAATTGGACCCAAATAACGCAGAAGCCTGGTACAATTTGGGTAACGCTTACTATAAGCAGGGT GACTATCAAAAAGCAATTGAAGACTACCAAAAGGCCTTAGAATTGGATCCTAATAACTTGCAAGC TGAAGCATGGAAAAATTTGGGTAACGCTTATTATAAACAGGGTGACTACCAAAAAGCCATTGAAT ACTATCAAAAAGCATTGGAATTGGATCCAAATAACGCCTCTGCTTGGTATAATTTGGGTAATGCTT ATTATAAGCAGGGTGACTACCAGAAAGCCATAGAATACTATCAAAAAGCTTTGGAATTAGACCCT AATAACGCAAAAGCCTGGTATAGACGTGGTAATGCTTACTACAAACAGGGTGACTATCAGAAGG CAATAGAAGATTATCAAAAAGCTTTAGAATTAGACCCAAATAACAGAAGTAGATCTGCTGGTGGT GGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTATGGAACCTGCAATGGAACCA GAAACATTGGAAGCCAGAATCAATAGAGCTACCAATCCTTTGAACAAGGAATTGGATTGGGCTTC TATTAATGGTTTCTGTGAACAATTGAACGAAGACTTCGAAGGTCCACCTTTAGCAACAAGATTATT GGCCCATAAAATTCAATCACCACAAGAATGGGAAGCAATACAAGCCTTAACCGTCTTGGAAACTT GTATGAAGTCCTGCGGTAAAAGATTCCACGATGAAGTTGGTAAATTCAGATTTTTGAACGAATTG ATCAAGGTTGTCTCACCTAAGTATTTGGGTAGTAGAACATCTGAAAAGGTTAAAAACAAGATCTT GGAATTGTTGTACTCCTGGACCGTAGGTTTACCAGAAGAAGTTAAGATCGCTGAAGCATACCAAA TGTTGAAGAAACAAGGTATTGTTAAGTCAGGTTCCGCCGGTAGTGCAGCCGGTTCTGGTGAATT CGGTTCTGCAGAAGCTGCAGCCAAGGAAGCTGCAGCCAAAGCTGGTTCAGCAGGTTCCGCTGC AGGTTCTGGTGAATTTGGTTCAGGTGCAATGGGTTCCATGGCCGAAGCTGAAGGTGAAAGTTTG GAATCTTGGTTAAATAAGGCTACAAATCCATCAAACAGACAAGAAGATTGGGAATATATCATTGG TTTCTGTGACCAAATCAATAAGGAATTGGAAGGTCCTCAAATAGCTGTTAGATTATTGGCACATAA GATCCAATCTCCACAAGAATGGGAAGCCTTACAAGCTTTGACTGTTTTAGAAGCTTGTATGAAGA ATTGCGGTAGAAGATTTCACAACGAAGTCGGTAAATTCAGATTTTTGAATGAATTAATTAAGGTAG TTAGTCCAAAATACTTAGGTGACAGAGTTTCTGAAAAGGTTAAGACCAAAGTCATAGAATTGTTGT ACTCTTGGACTATGGCCTTGCCTGAAGAAGCTAAGATCAAAGATGCATACCATATGTTGAAGAGA CAAGGTATAGTCCAATCAGATCCACCTATCCCAGTAGACAGAACTTTGATTCCATCTCCACCACC AAGACCTAAAAATGGTTCCGGT

4. Phosphomevalonate Kinase (ERG8) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID10 (SEQ ID NO:191)

ATGTCCGAATTAAGAGCTTTTAGTGCACCTGGTAAAGCCTTGTTAGCTGGTGGTTATTTGGTTTT GGATACAAAGTACGAAGCATTCGTTGTCGGTTTGTCAGCCAGAATGCATGCAGTCGCCCACCCT TACGGTTCTTTACAAGGTTCTGATAAGTTCGAAGTAAGAGTCAAGTCTAAGCAATTCAAGGACGG TGAATGGTTATACCATATATCTCCAAAGTCAGGTTTTATTCCTGTTTCCATAGGTGGTAGTAAAAA TCCATTCATCGAAAAGGTTATTGCAAACGTCTTTTCTTACTTCAAGCCTAACATGGATGACTACTG TAACAGAAACTTGTTCGTCATCGATATATTCTCTGATGACGCTTATCATTCTCAAGAAGACTCAGT

AACTGAACACAGAGGTAATAGAAGATTGTCCTTTCATAGTCACAGAATTGAAGAAGTTCCAAAAA CCGGTTTAGGTTCTTCAGCTGGTGGTTTAGTCACTGTATTGACTACAGCTTTAGCATCCTTTTTCG TTAGTGATTTGGAAAACAACGTAGACAAGTACAGAGAAGTTATTCATAATTTGGCACAAGTAGCC CACTGCCAAGCACAAGGTAAAATCGGTTCCGGTTTTGATGTTGCTGCAGCCGCTTATGGTTCAAT TAGATACAGAAGATTCCCACCTGCTTTGATATCTAATTTGCCAGATATCGGTTCTGCTACATATGG TTCAAAGTTGGCACATTTGGTTGATGAAGAAGACTGGAACATCACAATTAAATCCAACCATTTGC CTAGTGGTTTGACCTTATGGATGGGTGACATTAAGAATGGTTCTGAAACTGTTAAGTTGGTCCAA AAAGTAAAGAACTGGTACGATTCTCATATGCCAGAATCATTGAAGATCTACACAGAATTAGACCA TGCTAATTCCAGATTCATGGATGGTTTGAGTAAATTAGACAGATTGCATACCCACGATGACTACT CTGATCAAATCTTCGAATCATTGGAAAGAAACGACTGTACTTGCCAAAAATACCCAGAAATCACA GAAGTAAGAGATGCCGTTGCTACCATAAGAAGATCTTTTAGAAAGATCACTAAGGAATCAGGTGC AGATATCGAACCACCTGTTCAAACATCTTTGTTAGATGACTGTCAAACCTTGAAGGGTGTCTTAA CTTGCTTGATTCCAGGTGCTGGTGGTTATGATGCAATAGCCGTCATCACTAAACAAGATGTAGAC TTGAGAGCTCAAACAGCAAACGATAAGAGATTTTCAAAGGTCCAATGGTTAGATGTAACCCAAGC TGACTGGGGTGTTAGAAAAGAAAAGGATCCTGAAACTTACTTGGACAAAAAGTTATCTGGTGGTG GTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTACAATTTGGGTAA CGCATACTACAAGCAGGGTGACTACCAAAAGGCCATAGAATACTACCAAAAGGCTTTGGAATTG GACCCAAATAACGCCGAAGCTTGGTATAATTTGGGTAATGCTTATTATAAACAGGGTGACTATCA AAAGGCAATCGAATACTACCAAAAGGCCTTGGAATTAGACCCTAATAACGCAGAAGCCTGGTATA ATTTGGGTAACGCTTATTATAAGCAGGGTGACTATCAAAAAGCTATCGAAGACTACCAAAAGGCA TTGGAATTAGATCCAAATAACTTGCAAGCTGAAGCATGGAAGAATTTGGGTAACGCTTACTATAA ACAGGGTGACTACCAAAAAGCCATTGAATATTATCAAAAAGCTTTGGAATTGGATCCTAATAACG CCTCTGCTTGGTACAATTTGGGTAATGCTTACTATAAGCAGGGTGACTATCAGAAGGCTATTGAA TATTATCAAAAGGCTTTAGAATTGGACCCTAATAACGCAAAGGCCTGGTACAGACGTGGTAACGC TTATTACAAACAGGGTGACTACCAGAAAGCTATTGAAGATTATCAAAAGGCATTGGAATTGGATC CTAACAACAGATCCAGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTG GTGCTTCCAGTTATTACCATCACCATCACCATCACTTGGAATCTACATCATTATACAAAAAGGCTG GTTCCGGTAGTCAAAAGGTTGAAGAATTGAAAAATAAGATAGCCGAATTGGAAAACAGAAACGCT GTTAAAAAGAACAGAGTCGCACATTTGAAACAAGAAATAGCCTACTTGAAGGATGAATTAGCAGC CCATGAATTTGAAGGTTCTGCCGGTTCAGCTGCAGGTTCTGGTGAATTCGGTTCAGCTGAAGCC GCTGCAAAAGAAGCCGCTGCAAAGGCCGGTTCCGCTGGTAGTGCCGCTGGTTCTGGTGAATTT GGTTCTTCATACTATCACCATCACCATCATCACTTGGAATCTACTTCATTATATAAAAAGGCCGGT TCCGGTAGTTTCGAAAACGTTACACATGAATTCATTTTGGCTACCTTGGAAAACGAAAACGCAAA GTTAAGAAGATTGGAAGCCAAGTTGGAAAGAGAATTAGCTAGATTGAGAAATGAAGTTGCATGGT TAGGTTCTGGT

5. Diphosphomevalonate Decarboxylase (MVD1) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID11 (SEQ ID NO:192)

ATGACAGTTTATACCGCTTCTGTCACCGCACCTGTAAATATTGCTACTTTGAAATACTGGGGTAAA AGAGATACTAAGTTGAATTTGCCAACAAACTCTTCAATCTCAGTTACATTGTCCCAAGATGACTTA AGAACCTTGACTTCTGCTGCAACTGCTCCTGAATTCGAAAGAGATACATTGTGGTTGAATGGTGA ACCACATTCTATCGACAACGAAAGAACTCAAAACTGTTTGAGAGATTTGAGACAATTGAGAAAGG AAATGGAGAGTAAGGATGCTTCTTTGCCTACATTGAGTCAATGGAAGTTGCACATAGTTTCTGAA AACAACTTCCCAACCGCCGCTGGTTTGGCATCCAGTGCAGCCGGTTTCGCTGCATTAGTCTCTG CAATCGCCAAGTTGTACCAATTGCCACAAAGTACATCTGAAATCAGTAGAATCGCTAGAAAAGGT TCAGGTTCCGCATGTAGATCTTTATTTGGTGGTTACGTCGCATGGGAAATGGGTAAAGCCGAAG ACGGTCATGATTCAATGGCCGTACAAATAGCTGACTCTTCAGATTGGCCTCAAATGAAAGCTTGC GTCTTGGTTGTCTCAGACATCAAAAAGGATGTATCCAGTACACAAGGCATGCAATTGACTGTTGC AACATCCGAATTGTTTAAAGAAAGAATCGAACACGTAGTTCCAAAAAGATTCGAAGTCATGAGAA AGGCTATCGTAGAAAAGGATTTCGCCACCTTCGCTAAGGAAACTATGATGGACAGTAACTCTTTC CATGCAACTTGTTTGGATTCATTTCCACCTATTTTCTATATGAACGACACCTCAAAGAGAATAATC

TCCTGGTGCCACACTATCAACCAATTCTACGGTGAAACAATCGTTGCTTACACCTTCGATGCAGG TCCTAATGCCGTCTTGTATTACTTAGCCGAAAACGAATCAAAGTTGTTCGCTTTTATATATAAGTT GTTTGGTTCCGTTCCAGGTTGGGATAAAAAGTTCACTACAGAACAATTGGAAGCTTTTAATCATC AATTCGAATCTTCAAACTTTACTGCCAGAGAATTGGACTTAGAATTGCAAAAGGATGTAGCTAGA GTTATCTTGACCCAAGTTGGTTCAGGTCCTCAAGAAACTAACGAATCCTTGATAGATGCTAAGAC AGGTTTGCCAAAAGAAAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGT GGTAGTGCTGAAGCATGGTATAATTTGGGTAACGCTTATTACAAGCAGGGTGACTACCAAAAGG CCATCGAATACTACCAAAAGGCTTTGGAATTGGACCCTAATAACGCCGAAGCTTGGTACAATTTG GGTAATGCCTACTATAAACAGGGTGACTATCAAAAAGCAATTGAATATTACCAAAAGGCCTTGGA ATTGGACCCAAATAACGCAGAAGCCTGGTACAATTTGGGTAACGCCTACTATAAGCAGGGTGAC TATCAAAAGGCTATCGAAGATTACCAAAAGGCATTAGAATTGGATCCTAATAACTTGCAAGCTGA AGCATGGAAAAATTTGGGTAATGCCTATTATAAACAGGGTGACTACCAAAAAGCTATTGAATACT ATCAAAAAGCTTTAGAATTAGACCCAAATAACGCCTCAGCTTGGTATAATTTGGGTAATGCATACT ACAAACAGGGTGACTATCAGAAGGCAATTGAATACTATCAAAAGGCATTAGAATTAGATCCTAAT AACGCAAAAGCCTGGTATAGACGTGGTAATGCCTACTACAAGCAGGGTGACTATCAGAAGGCGA TTGAAGACTACCAAAAGGCATTGGAATTGGATCCAAACAACAGATCAAGATCCGCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGCAATGGCCGATTTGGAACAAAAG GTATTGGAAATGGAAGCTAGTACATATGACGGTGTTTTTATTTGGAAGATCTCTGATTTCCCAAGA AAAAGACAAGAAGCTGTTGCAGGTAGAATCCCTGCTATTTTTAGTCCAGCATTCTACACCTCTAG ATACGGTTACAAGATGTGTTTGAGAATATATTTGAATGGTGACGGTACTGGTAGAGGTACTCATT TGTCTTTGTTTTTCGTCGTAATGAAGGGTCCTAATGATGCTTTGTTGAGATGGCCTTTTAATCAAA AGGTTACCTTGATGTTGTTGGATCAAAACAACAGAGAACACGTTATCGACGCTTTTAGACCTGAT GTCACTTCCAGTTCTTTCCAAAGACCAGTTAATGATATGAACATTGCTTCTGGTTGTCCTTTGTTT TGCCCAGTCTCAAAGATGGAAGCTAAAAATTCCTATGTTAGAGATGACGCCATCTTCATTAAGGC TATCGTTGATTTGACTGGTTTAGGTTCAGCAGGTTCCGCCGCTGGTTCTGGTGAATTTGGTTCCG CCGAAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCAGGTAGTGCCGGTTCTGCAGCCGGCTCTG GCGAATTTGGTAGTGCCTCTATTAAATTGCAATCATCCGACGGTGAAATCTTCGAAGTTGATGTC GAAATAGCAAAGCAATCTGTTACCATAAAAACTATGTTGGAAGATTTGGGTATGGATGACGAAGG TGACGATGATCCAGTTCCTTTGCCAAATGTCAACGCTGCAATATTGAAGAAAGTTATTCAATGGT GCACACATCACAAGGACGATCCACCTCCACCTGAAGACGATGAAAATAAGGAAAAGAGAACTGA CGATATTCCAGTATGGGACCAAGAATTCTTGAAGGTTGATCAAGGTACATTGTTCGAATTGATCT TGGCCGCTAACTATTTGGACATCAAGGGTTTGTTAGATGTAACATGTAAAACCGTTGCTAACATG ATCAAGGGTAAAACACCAGAAGAAATCAGAAAGACCTTTAATATTAAGAATGATTTCACTGAAGAA GAAGAAGCACAAGTTAGAAAGGAAAACCAATGGTGCGGTTCTGGT

6. Isopentenyl-Diphosphate Delta-lsomerase (IDI1) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID12 (SEQ ID NO:193)

ATGACTGCTGATAATAACTCTATGCCACATGGTGCCGTATCTTCATACGCTAAGTTGGTTCAAAA CCAAACACCTGAAGATATCTTGGAAGAATTCCCAGAAATCATCCCTTTGCAACAAAGACCAAACA CTAGATCCAGTGAAACATCCAACGATGAAAGTGGTGAAACCTGTTTTTCAGGTCATGACGAAGAA CAAATTAAATTGATGAACGAAAACTGCATCGTATTGGATTGGGATGACAATGCAATAGGTGCCGG TACTAAGAAAGTTTGTCATTTGATGGAAAACATAGAAAAGGGTTTGTTGCACAGAGCTTTCTCCG TTTTTATATTCAATGAACAGGGTGAATTGTTATTGCAACAAAGAGCAACAGAAAAGATCACCTTTC CAGATTTGTGGACTAATACATGTTGCTCTCATCCTTTGTGCATTGATGACGAATTAGGTTTGAAGG GTAAATTGGATGACAAAATTAAGGGTGCTATAACTGCTGCAGTCAGAAAATTAGATCATGAATTG GGTATACCAGAAGACGAAACCAAGACTCGTGGTAAATTCCATTTCTTAAACAGAATCCACTATAT GGCTCCATCTAACGAACCTTGGGGTGAACATGAAATCGATTACATCTTATTTTACAAGATTAATGC AAAGGAAAACTTGACAGTTAACCCAAACGTTAATGAAGTCAGAGATTTCAAATGGGTTTCTCCTA ATGATTTGAAGACCATGTTTGCTGACCCATCATATAAGTTTACTCCTTGGTTCAAGATCATCTGTG AAAACTACTTGTTTAACTGGTGGGAACAATTAGATGACTTGTCTGAAGTTGAAAACGATAGACAA ATCCATAGAATGTTGAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTG

GTAGTGCCGAAGCTTGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAAAAGGC AATCGAATACTACCAAAAGGCCTTGGAATTGGACCCAAATAACGCAGAAGCCTGGTATAATTTGG GTAATGCATATTATAAACAGGGTGACTATCAAAAGGCTATTGAATATTACCAAAAGGCATTGGAAT TGGACCCTAATAACGCTGAAGCATGGTATAATTTGGGTAACGCCTATTATAAGCAGGGTGACTAT CAAAAAGCCATCGAAGACTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTACAAGCCGAAG CTTGGAAGAATTTGGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTATTGAATACTAT CAAAAAGCCTTAGAATTAGACCCTAATAACGCATCTGCCTGGTACAATTTGGGTAATGCCTATTA CAAGCAGGGTGACTATCAGAAGGCTATTGAATACTACCAAAAAGCATTGGAATTGGATCCAAATA ACGCTAAGGCATGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTATCAAAAGGCGAT TGAAGATTATCAAAAAGCTTTGGAATTGGATCCTAACAACAGATCTAGATCAGCTGGTGGTGGTG GTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCATATTACCATCACCATCACCATCAC TTAGAATCCACAAGTTTGTACAAAAAGGCTGGTTCTGGTTCAAACACCGTTAAGGAATTAAAGAA CTACATCCAAGAATTGGAAGAAAGAAACGCAGAATTGAAAAATTTGAAGGAACATTTGAAGTTTG CCAAGGCTGAATTAGAATTCGAATTGGCCGCTCACAAATTTGAAGGTTCCGCTGGTAGTGCAGC CGGTTCCGGTGAATTCGGTAGTGCAGAAGCTGCAGCCAAAGAAGCTGCAGCCAAGGCTGGTTC TGCAGGTTCAGCTGCAGGTTCTGGTGAATTTGGTTCCAGTTACTATCACCATCACCATCATCACT TAGAATCCACTAGTTTGTATAAAAAGGCCGGTTCTGGTTCACAAAAAGTCGCACAATTAAAGAAT AGAGTAGCCTACAAGTTGAAGGAAAACGCTAAGTTGGAAAACATTGTCGCAAGATTAGAAAACGA TAATGCCAACTTGGAAAAAGACATCGCTAATTTGGAAAAGGATATTGCAAACTTGGAAAGAGATG TTGCCAGAGGTTCTGGT

7. Geranyl-Diphosphate Synthase (ERG20ww) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID13 (SEQ ID NO:194)

ATGGAAGCTAAGATAGATGAATTGATAAATAACGACCCAGTTTGGTCTTCACAAAACGAATCCTT GATCAGTAAGCCATACAACCATATCTTGTTAAAACCTGGTAAAAATTTCAGATTAAATTTGATCGT ACAAATCAACAGAGTTATGAATTTGCCTAAGGATCAATTGGCTATCGTTTCTCAAATAGTCGAATT GTTGCATAACTCCAGTTTGTTGATCGATGACATCGAAGATAACGCACCATTGAGAAGAGGTCAAA CTACATCCCACTTAATTTGGGGTGTCCCTAGTACTATTAATACCGCAAACTACATGTACTTCAGAG CCATGCAATTGGTATCACAATTGACCACTAAGGAACCATTGTACCATTGGTTGATCACAATTTTTA ACGAAGAATTGATTAATTTGCACAGAGGTCAAGGTTTGGATATCTATTGGAGAGACTTCTTACCA GAAATTATACCTACCCAAGAAATGTACTTGAACATGGTAATGAATAAGACTGGTGGTTTGTTTAGA TTGACCTTGAGATTAATGGAAGCTTTGTCTCCATCTTCACATCACGGTCATTCATTGGTTCCTTTC ATAAACTTGTTGGGTATCATCTATCAAATCAGAGATGACTACTTGAATTTGAAGGATTTCCAAATG TCCAGTGAAAAGGGTTTCGCAGAAGACATAACTGAGGGTAAATTGTCATTCCCAATCGTCCATGC CTTAAACTTCACAAAAACCAAGGGTCAAACCGAACAACACAATGAAATCTTAAGAATTTTGTTATT GAGAACTTCTGATAAGGACATAAAGTTGAAGTTGATCCAAATCTTGGAATTCGATACCAACTCATT GGCTTACACTAAGAACTTCATCAACCAATTGGTTAACATGATTAAGAATGATAACGAAAATAAGTA CTTGCCAGATTTGGCCTCCCATAGTGACACTGCTACAAATTTGCACGATGAATTGTTGTACATCA TCGACCATTTGTCCGAATTGAAATTATCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGG TGGTGGTAGTGCAGAAGCCTGGTACAACTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAA AAGGCTATCGAATACTACCAAAAGGCATTGGAATTAGACCCAAATAACGCTGAAGCATGGTACAA CTTAGGCAACGCATATTATAAACAGGGTGACTATCAAAAGGCCATAGAATACTACCAAAAGGCTT TGGAATTGGACCCTAATAACGCCGAAGCTTGGTACAACTTGGGTAATGCTTATTACAAGCAGGGT GACTATCAAAAAGCAATTGAAGACTACCAAAAAGCCTTGGAATTAGATCCAAATAACTTGCAAGC AGAAGCCTGGAAGAACTTAGGCAACGCATACTATAAACAGGGTGACTACCAAAAAGCCATTGAA TATTATCAAAAAGCTTTGGAATTAGACCCTAATAACGCTTCTGCTTGGTATAACTTAGGCAATGCC TATTATAAGCAGGGTGACTATCAGAAAGCTATTGAATATTATCAAAAGGCCTTGGAATTGGACCC AAATAACGCCAAGGCTTGGTACAGACGTGGTAACGCATACTACAAACAGGGTGACTATCAGAAG GCTATCGAAGATTATCAAAAAGCATTAGAATTAGATCCTAATAACAGATCTAGATCAGCTGGTGG TGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTTGTGTACTATGAAAAAGGGT CCATCTGGTTACGGTTTTAATTTGCATTCTGATAAGTCAAAGCCTGGTCAATTCATAAGATCAGTT

GATCCAGACTCCCCTGCAGAAGCCAGTGGTTTGAGAGCTCAAGATAGAATTGTCGAAGTAAATG GTGTCTGCATGGAAGGTAAACAACACGGTGACGTTGTTTCTGCTATTAGAGCTGGTGGTGACGA AACTAAGTTATTGGTAGTTGACAGAGAAGGTTCCGCCGGTAGTGCTGCAGGTTCTGGTGAATTT GGTTCAGCTGAAGCCGCTGCAAAAGAAGCCGCTGCAAAGGCCGGTTCTGCTGGTTCAGCCGCT GGTTCTGGTGAATTCGGTTCTTCATCCGGTGCTATAATCTATACAGTTGAATTGAAGAGATACGG TGGTCCATTAGGTATTACTATATCTGGTACAGAAGAACCATTCGATCCTATCATCATCAGTTCTTT GACTAAGGGTGGTTTAGCTGAAAGAACAGGTGCAATCCATATTGGTGACAGAATATTGGCTATCA ATTCATCCAGTTTGAAAGGTAAACCATTGTCAGAAGCTATCCACTTATTGCAAATGGCAGGTGAA ACCGTTACTTTGAAAATCAAAAAGCAAACAGATGCACAACCTGCCTCTTCAGGTTCTGGT

Figure 14C

Codon-optimized CAN Gene Cassette Nucleotide Sequences

1. Olivetol Synthase (OS) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID14 (SEQ ID NO:195)

ATGAATCATTTGAGAGCCGAAGGACCAGCTTCTGTCTTAGCAATAGGTACTGCCAATCCAGAGAA CATCTTGTTACAAGATGAATTTCCTGACTATTACTTCAGAGTTACCAAATCCGAGCATATGACGCA GTTGAAGGAAAAGTTTAGAAAGATCTGTGATAAGAGTATGATCAGAAAGAGGAACTGCTTCTTAA ACGAAGAGCATTTGAAGCAAAATCCTAGATTAGTGGAACACGAGATGCAAACATTGGATGCTAG GCAGGACATGTTAGTTGTCGAAGTTCCTAAATTGGGTAAAGATGCATGTGCCAAAGCTATTAAGG AATGGGGTCAACCCAAGTCTAAGATAACTCATTTGATTTTTACTAGTGCTAGCACTACAGATATGC CTGGTGCAGACTATCACTGTGCCAAACTACTTGGTTTATCGCCCTCTGTGAAGAGAGTTATGATG TATCAACTAGGTTGCTACGGTGGTGGTACTGTACTTAGAATCGCTAAAGACATTGCAGAAAATAA CAAGGGTGCCAGGGTCTTGGCTGTATGTTGCGATATTATGGCTTGCTTGTTTAGAGGTCCATCA GAATCCGATTTGGAGCTGTTGGTTGGTCAAGCTATTTTCGGTGACGGTGCTGCAGCTGTTATTGT TGGTGCAGAACCTGATGAGTCAGTCGGTGAAAGACCAATCTTTGAATTGGTTTCTACCGGTCAAA CGATTTTACCAAATAGTGAAGGTACAATAGGTGGTCATATCAGAGAAGCTGGTTTGATATTCGAT TTGCACAAAGACGTTCCTATGCTAATATCTAACAACATCGAAAAGTGTCTGATCGAGGCTTTTAC CCCCATCGGTATTTCCGATTGGAATAGTATATTCTGGATCACGCATCCAGGTGGTAAAGCAATCC TGGATAAGGTTGAAGAGAAGCTGCATTTGAAGTCTGATAAGTTTGTCGACAGCAGACATGTATTG TCGGAACACGGTAACATGTCTTCATCCACAGTGCTGTTCGTTATGGATGAACTTAGAAAGAGATC TTTGGAAGAGGGTAAAAGCACCACGGGTGACGGTTTTGAATGGGGTGTTCTTTTTGGATTCGGC CCCGGTTTGACCGTCGAAAGAGTAGTTGTTAGATCTGTACCAATTAAATACAAGTTGTCTGGTGG TGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTACAATTTGGGT AACGCTTACTACAAGCAGGGTGACTACCAGAAGGCTATCGAGTATTACCAAAAAGCACTTGAACT GGATCCAAATAACGCTGAGGCATGGTATAATTTGGGCAACGCATATTACAAACAGGGTGACTATC AAAAGGCCATAGAATACTACCAAAAGGCTTTGGAGCTGGATCCTAATAACGCCGAAGCTTGGTA CAATTTGGGAAATGCCTATTATAAGCAGGGTGACTATCAGAAGGCAATAGAGGACTACCAAAAAG CCCTAGAACTTGATCCAAATAATTTGCAGGCAGAAGCCTGGAAGAATTTGGGTAATGCTTACTAT AAACAGGGTGACTATCAGAAAGCTATTGAATACTACCAAAAAGCACTGGAATTGGATCCTAATAA CGCTTCTGCTTGGTACAATTTGGGCAACGCTTACTACAAACAGGGTGACTACCAAAAAGCTATCG AATATTATCAAAAGGCTCTGGAACTAGATCCAAATAACGCCAAGGCTTGGTATAGAAGGGGAAAT GCTTATTATAAACAGGGTGACTACCAGAAAGCAATTGAAGACTACCAAAAAGCCCTTGAACTGGA TCCTAATAACAGATCTAGAAGCGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGT GGTGCTTCTGGTAACAACTTAGAAACATACGAGTGGTACAATAAGTCTATTTCTAGAGATAAGGC CGAAAAGTTACTACTTGACACCGGTAAAGAAGGTGCTTTTATGGTTAGAGATTCTAGAACTCCAG GTACTTATACAGTCTCTGTATTCACAAAGGCTATCATCTCAGAAAACCCATGTATCAAGCATTACC ACATCAAGGAAACCAACGACTCTCCTAAAAGATATTACGTGGCAGAAAAGTACGTTTTTGATTCA ATCCCACTGTTGATTCAATATCATCAGTACAATGGTGGTGGTTTGGTGACTAGATTGAGGTATCC TGTTTGCGGTGGTAGCGCAGGTTCGGCTGCAGGATCAGGCGAATTTGGTTCCGCCGAGGCCGC TGCAAAAGAAGCCGCTGCAAAGGCTGGATCTGCAGGCTCAGCCGCTGGTTCTGGAGAATTTGG TTCTGGTTCTCATCCCTGGTTTTTCGGTAAAATTCCAAGAGCAAAGGCCGAAGAAATGTTGTCTA AACAAAGACACGACGGTGCATTTTTGATAAGGGAAAGTGAGAGCGCACCTGGTGACTTTTCGTT GTCTGTTAAATTCGGTAATGATGTCCAACATTTCAAGGTATTGAGAGATGGTGCTGGTAAATACTT TTTGTGGGTCGTAAAGTTCAATTCCTTGAACGAATTAGTGGATTACCATAGATCAACTTCCGTTAG TAGGAACCAACAGATTTTCTTGAGAGATATCGAACAAGTTCCACAACAGCCTACAGGTTCTGGA

2. Olivetolic Acid Cyclase (OAC) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID15 (SEQ ID NO: 196)

ATGGCTGTAAAGCATTTGATCGTGTTGAAATTCAAGGATGAAATCACAGAGGCACAAAAGGAAGA GTTTTTCAAGACCTACGTTAATTTGGTCAACATAATCCCAGCTATGAAAGATGTATACTGGGGTAA AGACGTGACCCAAAAGAATAAGGAAGAGGGTTATACCCATATAGTAGAAGTGACGTTCGAATCA GTTGAAACTATCCAAGATTACATCATACACCCTGCTCATGTTGGCTTTGGTGACGTCTACAGATC CTTCTGGGAAAAGTTGCTGATCTTCGATTACACTCCAAGAAAGAAATTGTCTGGTGGTGGTGGTT

CTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTATAATTTGGGAAACGCTTA TTACAAACAGGGTGACTACCAAAAGGCCATCGAGTATTACCAAAAAGCTCTTGAACTGGACCCAA ATAACGCTGAGGCATGGTATAATTTGGGTAACGCATACTATAAGCAAGGTGACTACCAAAAGGCA ATTGAATATTACCAAAAGGCCTTGGAGTTAGACCCTAATAACGCCGAAGCTTGGTACAATTTGGG TAATGCCTACTATAAACAGGGTGACTATCAAAAGGCTATAGAGGACTACCAGAAAGCACTAGAAC TTGATCCCAATAACTTGCAAGCAGAAGCCTGGAAGAATTTGGGTAATGCCTATTATAAGCAAGGT GACTATCAAAAAGCTATTGAATACTACCAAAAAGCTCTGGAATTGGACCCTAATAACGCTTCTGC TTGGTATAATTTGGGTAATGCATACTACAAGCAAGGTGACTACCAGAAGGCAATAGAGTATTACC AAAAAGCCTTAGAACTAGACCCAAATAACGCCAAGGCTTGGTACAGAAGGGGTAATGCCTACTA CAAGCAGGGTGACTACCAAAAAGCTATTGAGGACTACCAAAAAGCACTTGAACTGGATCCTAATA ACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTC CGGTCAAGATAGAAGTGAAGCCACATTGATTAAAAGATTCAAAGGAGAAGGTGTTAGATACAAG GCTAAGCTGATCGGTATCGATGAAGTTTCTGCTGCTAGAGGTGACAAATTGTGTCAAGACTCTAT GATGAAGCTGAAGGGCGTTGTCGCAGGTGCCAGATCTAAGGGTGAACATAAGCAAAAGATATTT TTGACGATCTCATTCGGTGGTATTAAAATCTTCGATGAAAAGACTGGTGCTTTACAACATCACCAT GCAGTACACGAAATCTCTTACATCGCTAAGGATATCACAGACCATAGAGCATTCGGTTACGTTTG CGGTAAAGAAGGCAATCATAGATTTGTCGCTATTAAAACCGCCCAAGCCGCTGAACCAGTCATCT TGGATTTGAGAGACTTATTCCAGCTAATCTATGAACTAAAGCAAAGAGAAGAATTGGAAAAGAAA GCTGGTAGCGCAGGATCGGCAGCCGGTAGCGGAGAATTTGGTTCTGCTGAGGCTGCAGCCAAA GAAGCTGCAGCCAAGGCCGGCTCTGCTGGTTCAGCTGCAGGCTCTGGTGAATTTGGTTCTGGTT CTCATATGGGTTCTCAATTTTGGGTAACTTCTCAAAAGACTGAAGCTTCCGAGAGATGTGGTTTG CAAGGCTCCTATATTTTAAGGGTGGAAGCCGAGAAGCTTACCCTACTTACGCTGGGTGCACAGA GTCAAATATTGGAACCCCTGTTGTTCTGGCCATATACTTTATTGAGAAGATACGGTAGAGATAAA GTTATGTTCAGTTTCGAAGCTGGTAGAAGATGCCCAAGCGGTCCTGGAACTTTTACATTCCAGAC ATCACAAGGCAATGATATCTTTCAGGCAGTTGAAGCCGCTATTCAACAGCAAAAAGCCCAGGGT AAAGTCGGACAGGCTCAAGACATTCTAAGATTGGAACACCATCACCATCATCATGGTTCTGGT

3. CBGA Synthase (CBGAS) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID16 (SEQ ID NO:197)

ATGGGTTTGTCTTCAGTTTGTACATTCTCTTTCCAAACGAACTACCATACTTTGCTGAACCCTCAC AACAACAATCCCAAAACTTCTTTGCTTTGCTACAGACATCCAAAAACCCCTATTAAGTATAGCTAC AACAATTTCCCATCGAAACATTGTAGTACTAAGAGCTTCCATTTGCAAAATAAGTGCTCCGAATCT TTGTCTATCGCTAAGAACTCAATTAGAGCTGCAACTACAAATCAGACGGAACCACCTGAGTCGGA TAATCACTCTGTAGCCACCAAAATTTTGAACTTTGGTAAAGCTTGTTGGAAGCTGCAAAGAGCAT ACACAATAATAGCCTTCACCTCCTGTGCTTGCGGTTTGTTTGGTAAAGAACTGTTGCATAACACA AATTTGATTTCGTGGTCTTTGATGTTCAAGGCATTTTTCTTTTTGGTTGCAATCCTTTGCATCGCCT CTTTTACCACGACTATTAATCAAATCTATGATTTGCACATCGACAGAATTAATAAGCCCGATTTGC CACTAGCTTCAGGTGAAATCTCCGTTAATACTGCATGGATTATGTCAATCATTGTCGCCTTGTTCG GTTTAATCATCACAATTAAAATGAAAGGTGGTCCATTGTACATCTTCGGCTACTGTTTCGGTATAT TCGGTGGTATAGTATATTCCGTTCCACCTTTTAGATGGAAACAAAACCCCAGTACCGCTTTCTTAC TAAATTTCTTGGCACATATCATCACAAACTTCACCTTCTACTACGCTTCTAGAGCTGCTTTGGGTT TGCCATTCGAATTAAGACCATCTTTTACATTTTTGCTGGCTTTTATGAAATCGATGGGTTCTGCAT TGGCCTTGATTAAAGATGCATCTGACGTTGAAGGTGACACAAAATTCGGCATCAGTACCTTGGCT AGCAAGTACGGTTCTAGAAATTTGACTTTGTTTTGTTCAGGTATCGTATTGTTATCCTACGTGGCA GCCATTTTAGCCGGTATCATTTGGCCACAAGCTTTTAACAGTAATGTCATGCTACTTAGCCACGC AATATTGGCCTTCTGGCTGATCTTGCAGACGAGAGATTTTGCTTTAACTAATTATGACCCTGAGG CAGGTAGAAGATTCTACGAATTCATGTGGAAGCTGTACTACGCTGAATATTTGGTTTACGTCTTTA TTAAGTTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGC ATGGTACAACTTAGGCAACGCATACTACAAGCAGGGTGACTACCAGAAGGCAATTGAGTATTAG CAAAAAGCCTTAGAACTAGACCCAAACAATGCCGAGGCTTGGTATAACTTGGGCAATGCTTATTA CAAACAGGGTGACTATCAAAAGGCTATAGAATATTACCAAAAGGCACTTGAGCTGGACCCTAACA ATGCAGAAGCCTGGTATAACTTAGGCAATGCTTATTACAAGCAGGGTGACTATCAGAAGGCCAT

CGAGGACTACCAAAAGGCTTTGGAACTGGATCCAAACAATTTGCAGGCTGAAGCATGGAAGAAT TTGGGTAACGCTTACTATAAACAGGGTGACTATCAGAAAGCAATAGAATACTACCAAAAAGCCCT AGAACTTGACCCTAACAATGCCTCTGCTTGGTACAACTTGGGTAATGCTTACTATAAGCAGGGTG ACTACCAAAAAGCTATCGAATATTACCAAAAAGCACTGGAATTGGACCCAAACAATGCAAAGGCC TGGTATAGAAGAGGTAACGCCTACTACAAACAGGGTGACTACCAAAAGGCTATTGAAGATTACCA AAAGGCTCTGGAACTAGATCCTAACAACAGATCTAGATCCGCTGGTGGTGGTGGTTCTGGTGGT GGTGGTTCTGGTGGTGGTGGTGCTTCTGCAGAATACGTTAGAGCTCTGTTCGATTTCAACGGTA ACGATGAAGAGGACTTGCCTTTTAAGAAAGGTGACATTTTGAGAATCAGGGACAAACCAGAAGA GCAATGGTGGAATGCTGAAGATTCTGAGGGTAAAAGAGGAATGATTCCTGTTCCCTATGTCGAA AAGTACGGCTCAGCAGGTTCCGCTGCAGGATCTGGCGAATTCGGTTCAGCCGAGGCCGCTGCA AAAGAAGCCGCTGCAAAGGCTGGAAGTGCAGGCAGCGCCGCTGGTTCCGGAGAATTTGGTAGT TTGATTAAACATATGAGAGCCGAAGCTTTATTCGATTTTACTGGTAACTCCAAACTTGAACTGAAT TTCAAGGCAGGTGACGTTATTTTCTTGTTGAGTAGAATTAATAAGGACTGGTTGGAAGGTACTGT TAGAGGTGCTACTGGAATATTCCCACTTTCTTTTGTGAAAATCCTGAAGGGCTCAGGT

4. CBDA Synthase (CBDAS) (SEQ ID NO:198)

ATGAAATGTAGCACTTTTTCTTTCTGGTTCGTTTGCAAGATCATTTTCTTTTTCTTTTCTTTTAATAT CCAAACTTCGATCGCAAATCCAAGAGAAAACTTCTTAAAGTGTTTCTCACAATACATTCCTAATAA CGCCACGAATTTGAAGCTGGTATACACTCAGAACAACCCACTGTACATGAGCGTGCTAAACTCG ACAATCCATAATTTGAGATTCACTTCCGATACTACACCCAAACCATTAGTAATCGTGACACCTTCT CATGTTTCACACATTCAAGGAACCATACTATGCTCTAAGAAAGTCGGTTTGCAGATTAGAACAAG GTCTGGTGGTCATGATAGTGAAGGCATGTCCTACATCAGTCAAGTTCCATTCGTTATCGTCGATT TGAGAAACATGAGGTCTATCAAAATAGACGTTCACTCACAGACGGCTTGGGTCGAGGCAGGTGC CACTTTGGGAGAAGTTTACTACTGGGTCAACGAAAAGAATGAAAATTTGTCTCTTGCTGCAGGTT ACTGTCCAACTGTCTGCGCTGGTGGTCATTTTGGTGGTGGTGGTTATGGACCTCTTATGAGAAAC TACGGTTTGGCCGCTGATAATATCATTGACGCACATTTGGTAAATGTGCACGGTAAAGTTCTAGA TAGAAAGTCAATGGGTGAAGATTTGTTTTGGGCATTGAGAGGTGGTGGTGCTGAATCCTTTGGTA TAATCGTAGCTTGGAAAATTAGATTGGTTGCAGTCCCAAAGTCTACAATGTTCTCAGTTAAGAAAA TTATGGAAATCCATGAGCTGGTAAAGTTGGTGAATAAGTGGCAAAACATCGCTTACAAGTACGAT AAGGACTTGCTGCTAATGACCCATTTCATCACGAGAAACATCACTGATAACCAGGGTAAAAATAA GACAGCAATACACACCTACTTCTCTTCAGTTTTCTTGGGTGGTGTTGATTCCTTAGTGGATTTGAT GAATAAGAGTTTCCCTGAACTGGGTATTAAGAAAACTGATTGTAGACAATTGAGCTGGATCGACA CAATCATATTCTATAGTGGTGTTGTCAACTACGATACTGACAACTTCAACAAAGAAATCCTTCTGG ATAGAAGTGCCGGACAAAATGGCGCTTTCAAAATTAAGTTGGACTACGTTAAAAAGCCTATACCC GAGTCAGTATTTGTGCAGATCCTTGAAAAACTGTATGAAGAGGATATTGGTGCTGGAATGTACGC ATTATATCCATACGGTGGTATAATGGATGAAATCTCCGAGAGTGCCATACCATTCCCTCATAGAG CTGGTATCTTGTACGAACTGTGGTACATATGTTCTTGGGAAAAACAAGAGGATAACGAAAAGCAC TTAAACTGGATCAGGAACATCTATAACTTCATGACTCCTTACGTTTCTAAAAACCCCAGATTGGCT TATTTGAATTACAGAGATTTGGACATAGGTATCAACGATCCTAAAAATCCAAACAACTACACACAA GCAAGAATTTGGGGTGAAAAGTACTTCGGTAAAAATTTCGATAGATTGGTTAAAGTCAAGACCTT AGTTGACCCCAACAACTTTTTCAGAAACGAACAATCTATTCCACCTTTGCCTAGACATAGGCACG GCTCTGGT

5. CBCA Synthase (CBCAS) (SEQ ID NO:199)

ATGAACTGTAGCACTTTTTCTTTTTGGTTCGTTTGCAAGATAATATTTTTCTTTTTGTCCTTTAATAT CCAAATCAGTATCGCCAACCCACAGGAAAACTTTTTAAAGTGTTTCTCTGAGTACATCCCCAACA ACCCAGCTAACCCTAAGTTTATATATACACAACATGATCAGCTGTACATGAGCGTATTGAACTCG ACCATTCAAAATTTGAGATTCACTTCTGACACTACACCTAAGCCCTTGGTCATAGTAACTCCTTCT AATGTCTCACATATACAAGCTTCTATCTTGTGCTCTAAGAAAGTTGGTTTGCAGATTAGAACAAGG TCTGGTGGTCACGATGCAGAAGGTTTATCCTATATTAGTCAAGTCCCATTTGCCATAGTAGATTT GAGAAATATGCATACTGTGAAAGTTGACATACACTCACAGACTGCTTGGGTGGAAGCAGGTGCC

ACATTGGGAGAGGTTTACTACTGGATCAACGAGATGAACGAAAACTTTAGTTTCCCAGGTGGTTA CTGTCCCACAGTCGGTGTTGGTGGTCATTTTTCTGGTGGTGGTTATGGAGCTTTAATGAGAAACT ACGGTTTGGCTGCAGATAATATCATTGACGCACATTTGGTGAACGTTGATGGTAAAGTTCTTGAC AGAAAATCAATGGGTGAAGATTTGTTTTGGGCTATCAGAGGTGGTGGTGGTGAAAATTTCGGTAT AATCGCCGCTTGCAAAATTAAGTTGGTTGTCGTACCTAGCAAAGCTACTATTTTCTCTGTCAAAAA GAACATGGAAATCCATGGTTTAGTAAAGTTGTTTAATAAGTGGCAAAACATCGCATACAAGTACG ATAAGGATTTGATGCTTACCACGCATTTCAGAACTAGGAACATCACAGATAACCATGGTAAAAAT AAGACTACAGTTCACGGATACTTCTCTTCAATTTTCTTGGGTGGTGTTGATTCTCTTGTTGATTTG ATGAATAAGTCATTCCCAGAACTGGGTATTAAAAAGACAGATTGTAAGGAACTGAGCTGGATCGA CACCACGATTTTCTATAGTGGTGTGGTTAATTACAACACCGCCAACTTCAAAAAGGAAATCTTGC TGGATAGATCCGCTGGTAAAAAGACCGCTTTTTCTATTAAACTTGACTACGTTAAGAAACTGATCC CTGAAACTGCAATGGTTAAGATATTGGAGAAGCTGTACGAAGAGGAAGTCGGCGTAGGCATGTA CGTTTTGTATCCATACGGTGGTATAATGGATGAGATCTCCGAAAGTGCCATACCATTTCCTCATA GAGCTGGTATCATGTATGAATTATGGTACACCGCTACGTGGGAGAAGCAAGAAGATAACGAGAA ACACATAAACTGGGTCAGATCTGTATACAACTTCACTACACCTTACGTTTCTCAGAACCCAAGATT GGCATATTTGAACTACAGAGATTTGGACTTGGGTAAAACCAACCCCGAATCTCCAAATAACTATA CGCAAGCAAGAATTTGGGGTGAAAAGTACTTCGGTAAAAATTTCAACAGATTGGTGAAGGTTAAG ACAAAAGCCGATCCAAACAACTTCTTTAGAAACGAACAATCTATTCCACCATTGCCACCAAGACA TCATGGTTCCGGC

6. Acetyl-CoA Carboxylase (ACC) - Enzyme Linker - cTPR6 Spacer - ID Linker - ID17 (SEQ ID

NQ:200)

ATGTCAGAAGAGTCCTTATTTGAATCTTCACCACAAAAGATGGAGTACGAAATCACTAACTACTCT GAGAGACATACAGAATTGCCTGGACACTTCATCGGTTTGAACACAGTTGACAAGCTGGAAGAGT CTCCATTGAGAGATTTCGTCAAGTCCCATGGTGGTCACACCGTAATTAGTAAGATCTTGATAGCT AACAACGGTATCGCTGCAGTCAAGGAAATTAGATCTGTTAGAAAGTGGGCATATGAAACCTTTGG TGACGATAGAACGGTCCAATTCGTAGCTATGGCAACTCCTGAAGACTTGGAGGCCAATGCTGAA TATATCAGAATGGCCGATCAATACATTGAAGTTCCAGGTGGTACAAATAACAATAACTACGCTAAT GTCGACTTAATAGTAGATATCGCTGAAAGAGCAGACGTGGATGCCGTTTGGGCTGGTTGGGGAC ATGCTTCCGAAAACCCTTTGTTACCCGAAAAATTGTCTCAGAGTAAGAGAAAAGTTATTTTTATTG GTCCACCTGGAAATGCAATGAGATCATTAGGTGACAAGATATCCAGTACTATCGTGGCACAATCA GCCAAAGTTCCATGTATTCCTTGGTCCGGCACCGGTGTTGACACGGTGCATGTTGATGAAAAGA CTGGTTTGGTTTCTGTAGATGACGATATCTATCAGAAGGGATGTTGCACTTCACCTGAAGATGGT TTGCAAAAGGCTAAGAGAATCGGTTTCCCAGTTATGATCAAGGCATCAGAAGGTGGTGGTGGTA AAGGTATCAGGCAGGTCGAAAGAGAAGAGGATTTCATCGCTCTGTACCATCAAGCCGCTAATGA AATACCCGGTTCTCCAATTTTCATAATGAAACTAGCTGGAAGGGCAAGACATTTGGAAGTTCAGC TACTTGCTGACCAATACGGCACTAATATTTCCTTGTTCGGTAGAGATTGCAGTGTTCAAAGAAGA CATCAAAAGATTATCGAAGAGGCACCAGTCACTATAGCAAAAGCCGAAACATTTCACGAGATGGA AAAGGCAGCTGTTAGATTGGGTAAATTGGTCGGATATGTAAGTGCTGGAACAGTCGAATATTTGT ACAGCCATGACGATGGTAAATTCTACTTTTTGGAACTTAACCCAAGATTACAAGTTGAGCACCCT ACTACAGAAATGGTTTCTGGTGTTAATTTGCCAGCTGCACAACTGCAGATTGCTATGGGTATCCC TATGCATAGAATCAGTGATATCAGGACTCTGTACGGTATGAATCCACACAGCGCTTCGGAGATTG ACTTCGAATTCAAAACTCAGGATGCAACTAAGAAACAAAGAAGACCAATCCCAAAGGGTCATTGT ACCGCTTGCAGAATTACGTCCGAAGACCCCAATGATGGTTTTAAACCATCTGGTGGTACTTTGCA CGAACTAAACTTTAGAAGCTCGTCTAATGTCTGGGGTTATTTCTCAGTAGGCAACAACGGTAACA TCCATTCTTTTTCAGATTCCCAGTTCGGTCACATCTTCGCATTTGGAGAAAATAGGCAAGCCTCTA GAAAGCATATGGTTGTCGCTCTTAAAGAACTGTCAATCAGAGGTGACTTCAGAACCACGGTTGAA TACTTAATTAAACTGTTGGAAACTGAAGACTTCGAAGATAATACGATTACTACAGGTTGGTTGGAC GATTTGATAACCCATAAGATGACGGCAGAAAAACCTGATCCCACCTTGGCCGTTATCTGTGGTG CCGCTACGAAGGCCTTTTTAGCTTCTGAAGAGGCTAGACATAAGTACATAGAAAGCCTGCAAAA GGGTCAGGTACTATCGAAAGACTTACTACAAACAATGTTTCCTGTGGATTTCATCCACGAAGGTA

AAAGATACAAGTTTACTGTTGCTAAGTCTGGCAACGATAGGTACACGTTGTTCATTAATGGTAGC AAGTGCGACATCATTCTAAGACAACTTTCAGATGGTGGTTTGCTGATCGCAATTGGTGGTAAATC ACATACTATCTATTGGAAGGAAGAGGTCGCAGCCACAAGATTGAGTGTAGACAGCATGACCACG TTGTTAGAGGTTGAAAACGATCCAACTCAATTAAGAACACCATCTCCTGGTAAACTTGTGAAATTT CTGGTTGAAAATGGCGAGCATATAATCAAGGGTCAACCCTACGCTGAGATTGAAGTTATGAAAAT GCAGATGCCATTGGTTTCTCAAGAAAACGGTATAGTTCAACTACTTAAACAGCCTGGATCAACCA TAGTAGCTGGTGACATCATGGCAATTATGACGTTAGACGATCCATCCAAGGTGAAACATGCTCTT CCTTTTGAGGGTATGCTGCCCGATTTCGGTTCTCCAGTTATTGAAGGCACTAAACCAGCATACAA GTTTAAATCGTTGGTTTCTACACTGGAAAACATCCTAAAGGGTTACGATAACCAAGTTATTATGAA TGCTTCTTTGCAACAGTTGATAGAAGTCTTGAGAAATCCTAAGTTACCCTATTCAGAATGGAAATT GCATATTAGCGCTCTTCACTCGAGATTGCCTGCAAAATTGGATGAACAAATGGAAGAGCTAGTCG CTAGATCTTTGAGAAGAGGTGCTGTATTTCCAGCAAGGCAATTGAGTAAGCTAATTGACATGGCA GTTAAAAACCCAGAATACAACCCTGATAAACTGTTGGGTGCCGTAGTGGAACCATTGGCAGATAT TGCCCATAAGTACTCTAATGGTTTAGAAGCTCATGAGCACTCAATCTTCGTGCATTTCTTGGAAG AGTACTACGAGGTTGAAAAATTGTTCAACGGTCCTAACGTCAGAGAAGAGAACATCATCCTGAAG TTGAGAGATGAAAACCCAAAGGACTTGGATAAAGTCGCTCTTACTGTACTGAGTCATAGCAAGGT TTCTGCCAAAAATAACTTAATCCTAGCTATCCTGAAGCACTACCAACCTTTGTGTAAGCTGTCATC CAAAGTTTCTGCAATATTTTCAACTCCATTGCAACATATCGTAGAGCTTGAATCTAAGGCTACCGC AAAAGTGGCTTTGCAGGCAAGAGAAATTTTGATCCAAGGTGCTTTGCCATCAGTTAAAGAAAGAA CAGAGCAAATAGAACACATCCTGAAGAGTAGCGTTGTCAAAGTCGCATACGGTTCGTCTAATCCT AAGAGATCTGAACCCGATTTGAATATACTTAAGGATTTGATCGATTCAAATTACGTAGTGTTTGAC GTTTTACTACAGTTCTTAACTCATCAAGATCCTGTTGTCACAGCTGCAGCCGCTCAAGTCTATATA AGAAGGGCCTATAGAGCTTACACTATCGGTGACATTAGGGTACACGAAGGCGTGACAGTTCCAA TCGTGGAATGGAAATTTCAATTGCCCTCCGCAGCCTTTAGTACCTTCCCAACGGTAAAGTCAAAA ATGGGTATGAACAGAGCTGTTTCTGTTTCTGATTTGAGCTATGTGGCTAATTCGCAATCATCCCC TTTAAGAGAAGGTATTCTAATGGCTGTGGACCATTTGGACGATGTTGATGAAATTTTGTCTCAATC TTTGGAAGTTATTCCAAGACACCAAAGTAGCTCGAATGGTCCCGCTCCAGATAGGTCTGGATCTT CAGCAAGTTTAAGCAACGTAGCCAATGTGTGTGTTGCTTCCACTGAGGGTTTTGAAAGTGAAGA GGAAATCTTGGTTAGATTGAGAGAAATTTTGGATTTGAACAAACAAGAATTGATTAATGCTTCCAT CAGAAGGATCACATTCATGTTCGGTTTTAAAGATGGTAGTTACCCTAAGTACTACACCTTTAATGG TCCCAACTACAACGAGAACGAAACTATCAGACATATCGAACCTGCCTTAGCTTTCCAATTGGAAC TGGGTAGATTGTCAAACTTCAACATCAAGCCAATTTTCACTGATAACAGAAACATCCATGTGTAC GAAGCTGTTTCAAAGACATCCCCATTAGATAAGAGATTTTTCACCAGAGGCATCATTAGGACGGG TCACATTAGAGATGATATTAGCATACAAGAGTACTTGACTTCGGAAGCTAACAGATTAATGTCTGA CATCCTAGATAATTTGGAAGTTACCGACACGTCGAACTCTGATTTGAACCATATCTTTATTAACTT CATCGCAGTGTTCGACATATCTCCTGAGGATGTTGAAGCTGCATTTGGTGGTTTCTTGGAAAGAT TCGGTAAAAGATTGCTGAGATTGAGAGTCTCCAGTGCTGAAATCAGAATCATCATTAAGGATCCA CAAACTGGTGCCCCTGTACCCCTGAGAGCTTTGATCAATAATGTTTCTGGTTACGTAATTAAAAC CGAGATGTACACGGAAGTCAAGAATGCTAAGGGTGAATGGGTATTCAAGAGCTTGGGTAAACCC GGCTCGATGCACTTAAGACCAATTGCAACACCATATCCTGTCAAAGAATGGTTGCAACCTAAGAG ATACAAAGCCCACTTAATGGGTACTACATACGTTTACGATTTCCCAGAATTGTTCAGACAGGCTT CTTCTTCTCAATGGAAGAATTTTTCCGCCGACGTTAAGCTGACTGACGATTTCTTTATCAGTAACG AACTAATCGAGGATGAAAATGGTGAACTTACAGAGGTTGAAAGAGAGCCAGGAGCAAATGCCAT TGGCATGGTCGCTTTTAAGATCACTGTAAAGACACCAGAATATCCTAGGGGTAGACAATTCGTAG TGGTTGCAAACGACATCACCTTTAAAATTGGTTCTTTCGGACCTCAAGAAGATGAGTTTTTCAATA AGGTTACTGAATACGCTAGGAAAAGAGGTATACCAAGAATCTACTTGGCCGCTAATTCTGGAGCA AGGATTGGCATGGCCGAGGAAATAGTGCCTTTATTTCAGGTTGCATGGAACGACGCAGCCAACC CAGATAAGGGATTCCAATATTTGTATTTGACTTCTGAGGGTATGGAAACATTGAAAAAGTTCGATA AGGAAAACTCAGTGCTGACCGAGAGAACTGTTATTAATGGAGAGGAAAGGTTCGTAATCAAAACT ATAATCGGTTCTGAAGATGGTTTGGGCGTGGAGTGTCTGAGAGGTAGCGGTTTGATTGCTGGTG CAACTTCTAGAGCTTACCATGATATTTTTACTATCACACTGGTCACTTGCAGATCTGTAGGCATAG GTGCTTATTTGGTTAGATTGGGTCAAAGGGCCATCCAGGTCGAAGGCCAACCTATTATATTGACT

GGTGCCCCCGCTATAAACAAAATGCTGGGTAGAGAAGTTTATACCTCCAATTTGCAGTTGGGTG GTACGCAAATCATGTACAATAACGGTGTTTCTCATTTGACAGCTGTAGACGATTTGGCTGGTGTG GAAAAGATTGTTGAATGGATGTCATATGTGCCAGCTAAAAGAAACATGCCCGTTCCAATATTGGA AACTAAGGACACATGGGATAGACCAGTAGATTTTACCCCTACGAATGACGAAACCTATGATGTGA GATGGATGATTGAGGGTAGGGAAACTGAGTCTGGTTTTGAATACGGTTTGTTCGATAAGGGTTCT TTCTTTGAAACATTATCAGGCTGGGCCAAGGGTGTCGTAGTGGGAAGAGCTAGATTGGGTGGTA TTCCTCTAGGTGTTATTGGTGTAGAAACTAGAACAGTTGAAAATTTGATCCCCGCAGATCCAGCC AACCCTAATTCTGCTGAAACTTTAATTCAGGAACCTGGTCAAGTTTGGCATCCCAACTCAGCTTTT AAAACCGCACAGGCCATTAATGATTTCAACAACGGTGAACAATTGCCAATGATGATACTGGCTAA CTGGAGAGGTTTTTCTGGTGGTCAAAGGGATATGTTCAACGAAGTTTTGAAGTACGGTAGTTTTA TCGTCGACGCACTGGTAGATTACAAGCAACCTATCATAATATACATTCCACCAACTGGTGAATTA AGAGGTGGTTCTTGGGTTGTCGTAGACCCAACCATTAACGCAGATCAGATGGAAATGTACGCCG ATGTGAATGCTAGAGCAGGTGTTTTGGAACCACAAGGAATGGTTGGTATTAAGTTTAGAAGAGAA AAATTGCTGGATACTATGAACAGATTAGACGATAAGTACAGGGAATTGAGATCTCAACTGAGCAA TAAGTCTTTGGCTCCAGAAGTTCATCAACAGATCTCTAAGCAACTGGCTGATAGGGAAAGAGAAT TGTTGCCAATATACGGTCAGATCTCATTGCAATTTGCCGACTTACACGATAGGTCATCCAGAATG GTGGCTAAGGGTGTTATTTCAAAAGAATTAGAGTGGACAGAAGCTAGAAGATTTTTCTTTTGGAG ATTGAGAAGAAGATTGAACGAGGAATATTTGATTAAAAGATTGTCACATCAAGTTGGCGAGGCTT CTAGATTGGAAAAGATCGCAAGGATTAGATCTTGGTATCCAGCATCAGTCGATCACGAAGACGAT AGACAAGTAGCCACTTGGATTGAGGAAAATTACAAGACACTGGACGATAAGTTGAAGGGTTTAAA GCTAGAATCCTTTGCCCAAGACTTGGCTAAAAAGATTAGAAGTGACCATGATAATGCTATCGATG GTTTGAGTGAAGTTATTAAAATGCTTAGCACTGACGATAAGGAAAAACTGTTGAAGACATTGAAG AAACTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCT TGGTATAACTTGGGAAATGCTTATTACAAGCAGGGTGACTACCAAAAGGCCATAGAATACTACCA AAAGGCTCTTGAGCTGGATCCTAATAACGCAGAAGCCTGGTATAACTTAGGCAATGCATACTATA AACAAGGTGACTACCAAAAGGCAATAGAGTACTACCAAAAGGCCTTGGAATTAGATCCAAATAAC GCTGAGGCATGGTATAACTTGGGCAACGCCTACTATAAACAGGGTGACTATCAAAAGGCTATAG AAGATTACCAGAAGGCACTAGAGCTTGATCCTAATAACTTGCAAGCCGAAGCTTGGAAGAACTTA GGAAATGCATACTATAAGCAAGGTGACTATCAAAAAGCTATTGAATATTACCAAAAGGCTCTGGA GTTGGATCCAAATAACGCATCTGCTTGGTACAACTTAGGCAACGCCTACTATAAGCAGGGTGACT ATCAAAAAGCAATTGAATATTATCAAAAGGCCTTAGAGCTAGATCCTAATAACGCTAAAGCATGGT ATAGGAGAGGCAATGCATACTACAAACAGGGTGACTACCAAAAAGCTATAGAAGATTACCAAAAG GCACTTGAACTGGATCCAAATAACAGATCTAGATCCGCTGGTGGTGGTGGTTCTGGTGGTGGTG GTTCTGGTGGTGGTGGTGCTTCTGGTTCTCATATGAGATTGGGAGCCCAATCTATTCAGCCAAC CGCTAACTTAGATAGAACGGACGATTTGGTCTATTTGAATGTAATGGAATTGGTTAGAGCTGTTTT GGAGTTGAAAAATGAACTAGCACAATTGCCACCAGAAGGTTACGTGGTTGTCGTAAAGAATGTTG GTTTGACTCTTAGAAAGTTGATAGGCTCGGTCGACGATTTGCTACCATCTTTGCCATCTTCTTCTA GAACTGAAATAGAGGGTACACAAAAGCTTCTGAACAAAGATTTGGCTGAATTGATTAATAAGATG AGATTGGCACAACAGAACGCCGTTACTTCTTTGTCTGAGGAGTGTAAGAGACAAATGCTAACTGC TTCTCATACTTTGGCTGTTGATGCAAAGAACTTGTTAGACGCTGTGGATCAAGCAAAAGTTTTAG CCAATTTGGCTCACCCACCTGCCGAAGGTTCTGCTGGATCAGCTGCAGGATCCGGCGAATTTGG TTCTGCTGAAGCCGCTGCAAAAGAGGCTGCTGCAAAAGCTGGATCTGCAGGTAGTGCTGCTGGT AGCGGAGAATTTGGTTCTGGTGCCATGGCTACTCCTGGTTCAGAAAACGTTCTACCAAGAGAAC CATTGATTGCAACAGCCGTGAAGTTCTTGCAGAACTCTAGAGTTAGACAATCTCCATTGGCAACT AGAAGAGCATTTTTGAAAAAGAAAGGTTTGACCGACGAGGAAATTGATATGGCTTTCCAACAGTC CGGTACTGCAGCCGATGAACCATCTTCATTGTGGGGAAGTGGC

Figure 14D

Codon-optimized SCF Gene Cassette Nucleotide Sequences

1. Cannabinoiderqic Metabolon Scaffold (CBSCFLD) - (Myc)3

ATGGGTTCTGCTGGTTCAGCTGCAGGTTCTGGTGAATTCGGTTCCGCTGGTAGTGCCGCTGGTT CTGGTGAATTTGGTTCTGCTGGTTCAGCAGCCGGTTCTGGTGAATTCTCCTATTACCATCACCAT CACCATCACTTGGAATCTACTTCATTATACAAAAAGGCTGGTTCCGGTAGTGCCAGAAACGCTTA CTTGAGAAAGAAAATTGCTAGATTGAAGAAAGATAATTTGCAATTGGAAAGAGATGAACAAAACTT GGAAAAGATTATCGCTAATTTGAGAGATGAAATAGCAAGATTGGAAAATGAAGTTGCTTCTCATG AACAAGGTTCCGCAGGTAGTGCCGCCGGTTCTGGTGAATTTGCTGAAGCCGCTGCAAAGGAAG CCGCTGCAAAAGCAGGTTCTGCCGGTTCAGCCGCTGGTAGTGGTGAATTTTCTTACTATCACCAT CACCATCATCACTTGGAATCTACCTCATTATATAAAAAGGCCGGTTCCGGTAGTAACTTGGTTGC TCAATTAGAAAATGAAGTCGCATCATTGGAAAACGAAAACGAAACTTTGAAAAAGAAAAACTTACA TAAGAAAGATTTGATCGCTTACTTAGAAAAGGAAATAGCAAATTTGAGAAAGAAAATAGAAGAAG GTTCCGCTGGTAGTGCAGCCGGTAGTGGTGAATTCGGTTCTGCTGAAGCTGCAGCCAAGGAAG CTGCAGCCAAAGAAGCCGCTGCTAAAGAAGCTGCAGCCAAAGCTGGTTCTGCAGGTTCTGCCG CAGGTTCCGGTGAATTTGGTTCTTCATACTATCACCATCACCACCACCACTTGGAATCTACCTCA TTATACAAGAAAGCTGGTTCCGGTAGTCAAAAGGTCGCTGAATTGAAAAACAGAGTAGCTGTTAA GTTGAACAGAAACGAACAATTGAAAAATAAGGTAGAAGAATTGAAAAATAGAAACGCTTACTTGA AAAACGAATTGGCAACTTTGGAAAACGAAGTAGCTAGATTAGAAAACGATGTTGCTGAAGGTTCT GCTGGTTCTGCTGCTGGTTCTGGTGAATTCGCTGAAGCAGCCGCTAAGGAAGCAGCCGCTAAA GCCGGTTCCGCCGGTTCTGCTGCGGGCTCTGGTGAATTTTCCTACTATCACCATCATCATCACC ACTTGGAATCTACATCATTATATAAGAAAGCCGGTTCCGGTAGTAATGAAGTTACTACATTGGAAA ACGATGCTGCTTTTATTGAAAACGAAAACGCATACTTAGAAAAGGAAATCGCTAGATTGAGAAAG GAAAAGGCCGCTTTGAGAAATAGATTAGCTCATAAGAAAGGTTCTGCTGGTAGCGCTGCTGGCT CTGGTGAATTTGGTTCCGCCGAAGCCGCTGCTAAGGAAGCCGCTGCCAAAGAAGCCGCTGCCA AGGAAGCCGCTGCTAAGGCTGGTTCCGCCGGTTCAGCTGCAGGCTCTGGTGAATTCGGTTCTA GACCACCTACCATCTCTAATCCACCTCCATTGATTTCCAGTGCTAAACATCCATCCGTCGGTAGT GCAGGTTCCGCTGCCGGCTCTGGCGAATTTGCCGAAGCTGCTGCCAAAGAAGCAGCCGCTAAA GCTGGTTCAGCAGGTTCCGCTGCCGGATCTGGCGAATTCAATTTCTTGCAATCTAGACCAGAAC CTACTGCTCCTCCAGAAGAAAGTTTCAGATCTGGTGGTTCAGCTGGTTCCGCCGCAGGATCTGG CGAATTTGGTTCCGCAGAAGCTGCCGCTAAAGAAGCTGCTGCAAAAGAAGCAGCCGCCAAAGAA GCTGCTGCAAAAGCCGGTAGTGCTGGTTCAGCTGCCGGTTCCGGTGAATTCGGTTCTTCAAAAG GTACCGGTTTAAATCCAAACGCTAAAGTTTGGCAAGAAATTGCTCCTGGTAACGGTTCTGCAGGT TCCGCAGCTGGTTCCGGTGAATTCGCCGAGGCCGCTGCTAAGGAAGCAGCAGCCAAAGCAGGT AGTGCTGGTTCCGCAGCTGGTTCAGGTGAATTCCCAGACGGTGGTACCACTTTCGAACATTTGT GGTCCAGTTTAGAACCTGATTCTACATACGGTTCTGCCGGTTCTGCAGCAGGCAGCGGTGAATT CGGTTCTGCCGAAGCTGCTGCTAAAGAAGCTGCTGCCAAGGAAGCTGCTGCTAAGGAAGCTGC TGCCAAAGCCGGTAGTGCAGGTTCTGCTGCCGGTTCAGGTGAATTTGGTTCTTCTTACTATCACC ACCACCACCATCACTTGGAATCTACATCATTATACAAGAAAGCCGGTTCTGGTAGTAAGAGAATC GCATACTTAAGAAAGAAAATCGCTGCATTGAAGAAAGATAACGCAAACTTAGAAAAGGACATCGC TAACTTGGAAAACGAAATCGAAAGATTGATTAAAGAAATCAAAACCTTGGAAAATGAAGTTGCATC TCATGAACAAGGTTCAGCCGGTTCTGCAGCGGGCTCCGGTGAATTTGCCGAAGCTGCAGCAAAA GAAGCTGCCGCTAAGGCTGGTAGTGCTGGTTCTGCTGCAGGCAGCGGTGAATTTTCTTACTACC ACCATCACCACCATCACTTGGAATCTACTTCATTATATAAGAAAGCAGGTTCTGGTAGTAACTTGT TAGCAACATTAAGATCTACCGCTGCAGTCTTGGAAAACGAAAACCATGTATTGGAAAAAGAAAAG GAAAAATTGAGAAAGGAAAAAGAACAATTGTTGAATAAGTTGGAAGCTTACAAAGGTTCAGCAGG TTCTGCAGCGGGCTCTGGCGAATTCGGTTCCGCCGAAGCTGCAGCAAAGGAAGCTGCAGCTAA AGAGGCCGCTGCAAAAGAAGCTGCTGCCAAAGCAGGTAGTGCAGGTTCCGCAGCCGGCTCCG GCGAATTTGGTTCACCAGCTACATCCCAACATCCTCCACCTCCACCTGGTCATAGATCTCAAGCT CCTTCACATGGTTCCGCAGGTAGTGCAGCTGGATCTGGCGAATTCGCCGAAGCTGCCGCTAAG GAAGCTGCTGCAAAAGCTGGTTCCGCTGGTTCAGCAGCAGGTTCCGGTGAATTCGAATTGAATT

CTTTGTTGATATTGTTAGAAGCAGCCGAATATTTGGAAAGAAGAGATAGAGGTTCTGCCGGTAGT GCTGCAGGTAGCGGCGAATTTGGTTCTGCAGAAGCAGCCGCCAAGGAAGCAGCTGCAAAAGAA GCAGCAGCTAAAGAAGCAGCTGCAAAAGCCGGTTCTGCTGGTTCAGCCGCAGGATCTGGAGAA TTCGGTTCCAGACCACCTACAATTTCCAATCCACCTCCATTGATCTCTTCTGCCAAGCATCCATC CGTTGGTAGTGCAGGTTCAGCTGCCGGTAGTGGTGAATTTGCCGAAGCCGCCGCTAAGGAAGC CGCCGCCAAAGCAGGTTCAGCCGGTTCCGCCGCAGGTTCAGGTGAATTCAATTTCTTGCAGTCA AGACCAGAACCTACCGCTCCTCCAGAGGAGAGTTTCAGATCTGGTGGTAGTGCCGGTTCAGCTG CCGGCTCTGGAGAATTTGGTTCTGCAGAGGCTGCTGCCAAGGAAGCCGCAGCTAAAGAAGCCG CTGCGAAAGAAGCCGCCGCTAAAGCTGGTAGTGCAGGTAGTGCTGCGGGATCTGGCGAATTCG GTTCTTCTAAGGGTACTGGTTTGAACCCTAATGCCAAGGTCTGGCAAGAAATCGCCCCTGGTAA CGGTTCCGCAGGTTCCGCCGCAGGTAGTGGTGAATTCGCCGAGGCTGCCGCCAAGGAAGCCG CCGCTAAGGCAGGTAGTGCTGGTTCAGCGGCCGGCTCTGGTGAATTTCCAGACGGTGGTACAA CCTTTGAGCATTTGTGGTCCAGTTTAGAACCTGATTCTACGTACGGTTCTGCTGGTTCCGCTGCA GGATCTGGCGAATTCGGTTCCGCGGAAGCCGCCGCAAAAGAAGCCGCCGCCAAAGAAGCCGC CGCAAAGGAAGCCGCAGCAAAGGCAGGTAGTGCCGGCTCCGCCGCTGGCAGTGGCGAATTTG GTTCTTCATATTATCACCATCATCATCATCACTTGGAATCTACTTCATTATACAAGAAAGCAGGTT CCGGTTCTAAAAGAATTGCTTACTTAAGAAAGAAAATCGCGGCTTTGAAGAAAGACAATGCTAAC TTAGAAAAAGATATTGCCAACTTGGAAAATGAAATCGAAAGATTAATTAAGGAAATTAAAACATTG GAAAACGAAGTTGCATCACATGAACAAGGTTCAGCTGGTTCCGCTGCAGGGTCCGGCGAATTTG CAGAAGCCGCCGCCAAGGAAGCCGCAGCCAAAGCTGGTAGTGCAGGTTCTGCCGCTGGCTCTG GCGAATTTTCTTACTATCATCATCACCATCACCACTTGGAATCTACTTCATTATACAAGAAAGCGG GTTCAGGTTCTAACTTGTTAGCAACTTTAAGATCTACAGCCGCTGTTTTAGAAAATGAAAACCATG TCTTAGAAAAAGAAAAGGAAAAGTTGAGAAAGGAAAAGGAACAATTATTAAATAAGTTAGAAGCC TACAAGGGTTCAGCAGGTTCCGCAGCAGGCTCAGGCGAATTTGGTTCTGCAGAAGCGGCTGCT AAGGAAGCTGCCGCAAAGGAAGCAGCTGCTAAGGAGGCCGCTGCAAAGGCTGGTTCTGCTGGT TCCGCCGCGGGCTCTGGAGAATTCGGTTCCGCTTTGGTTGATGACGCCGCTGATTATGAACCTC CACCTTCAAATAACGAAGAAGCTTTAGGTTCCGCTGGTTCCGCTGCAGGTTCCGGCGAGTTCGC AGAAGCCGCAGCAAAAGAAGCCGCAGCTAAGGCAGGTAGTGCCGGATCCGCCGCTGGCAGTG GAGAATTCAGAGAATTGTTCGATGACCCATCTTACGTCAACGTACAAAATTTGGATAAAGCTAGA CAAGGTTCCGCCGGTTCTGCAGCGGGATCTGGGGAATTTGGTTCTGCAGAAGCTGCCGCCAAA GAAGCTGCAGCTAAAGAAGCCGCAGCCAAAGAAGCTGCTGCTAAGGCCGGTTCTGCTGGTTCT GCCGCAGGATCTGGGGAATTCGGTTCCAAGAATACTAAGAGTATGAACTTCGATAACCCAGTTTA CAGAAAGACTACAGAAGAAGAAGGTTCAGCCGGTTCAGCCGCCGGTTCCGGTGAATTTGCAGA GGCTGCCGCTAAAGAGGCTGCCGCTAAGGCCGGTAGTGCTGGTTCTGCAGCCGGCTCCGGAG AATTCAGATCTTTGCCATCCACATGGATTGAAAACAAATTATACGGCATGTCAGACCCTAATTGG GGTTCTGCAGGTTCAGCTGCGGGATCTGGTGAATTCGGTTCAGCAGAAGCCGCAGCCAAGGAA GCCGCTGCAAAGGAGGCCGCTGCCAAAGAAGCAGCTGCTAAGGCTGGTTCAGCCGGTTCCGCA GCCGGCAGTGGTGAATTTGGTAGTGTTGTCGATAATTCTCCACCTCCAGCTTTGCCTCCAAAGAA AAGACAATCTGCTCCATCTGGTTCAGCAGGTTCAGCCGCTGGTTCAGGTGAATTTGCCGAAGCA GCTGCCAAGGAAGCTGCCGCCAAGGCGGGCAGTGCAGGTTCGGCTGCGGGGTCTGGTGAATT CACTCAAAGATCTAAACCACAACCTGCAGTTCCTCCAAGACCATCTGCTGACTTGATTTTAGGTT CCGCCGGTTCCGCAGCTGGCTCTGGCGAATTCGGTTCCGCTGAGGCTGCCGCTAAAGAAGCGG CCGCTAAAGAGGCAGCCGCTAAAGAGGCGGCCGCTAAAGCAGGTTCTGCAGGTTCAGCAGCAG GTAGTGGTGAATTTGGTTCTACAGATGAAGAAAGAGAAGAAACCGAAGAAGAAGTTTATTTGTTG AACTCTACCACTTTGGGTTCAGCTGGTTCTGCTGCGGGTTCTGGCGAATTTGCAGAAGCAGCTG CTAAGGAAGCCGCGGCAAAGGCTGGTTCTGCGGGCTCCGCCGCAGGTTCTGGTGAATTTGATG GTAATGTATCTGGTACTCAAAGATTAGACTCAGCTACCGTTAGAACTTATTCATGCGGTTCTGCC GGTAGTGCAGCGGGCTCTGGGGAATTCGGTTCCGCAGAAGCCGCTGCAAAAGAAGCCGCTGCA AAAGAAGCCGCTGCGAAGGAGGCTGCTGCTAAGGCAGGTTCCGCCGGTAGTGCTGCGGGTTCC GGCGAATTTGGTTCCAGTTACTATCACCATCATCACCACCACTTGGAATCCACAAGTTTATATAA GAAAGCTGGTTCTGGTTCACAAAAGGTAGCTCAATTGAAAAATAGAGTTGCATACAAGTTGAAGG AAAACGCTAAGTTGGAAAACATAGTAGCAAGATTAGAAAACGATAACGCTAATTTGGAAAAGGAC

ATCGCAAATTTGGAAAAGGATATAGCTAACTTGGAAAGAGATGTTGCTAGAGGTTCTGCTGGTAG TGCCGCAGGCTCTGGCGAATTCGCTGAAGCTGCCGCTAAAGAGGCTGCGGCTAAAGCTGGTTC AGCTGGTTCTGCAGCGGGGTCTGGTGAATTTTCTTATTATCACCATCATCACCATCACTTGGAAT CCACCAGTTTATACAAGAAAGCCGGCTCTGGTTCAAACACTGTTAAGGAATTGAAAAATTACATT CAAGAATTGGAAGAAAGAAACGCTGAATTGAAAAATTTGAAGGAACATTTGAAGTTTGCAAAAGC CGAATTGGAATTCGAATTAGCAGCCCATAAATTTGAAGGTTCTGCCGGTTCTGCCGCCGGATCT GGAGAATTTGGTTCTGCGGAGGCTGCCGCTAAAGAAGCCGCCGCTAAAGAGGCTGCAGCTAAG GAAGCTGCAGCAAAGGCTGGTTCTGCCGGTTCCGCTGCCGGCTCCGGCGAATTTGGTTCACAT GATGACTCCTTGCCACATCCTCAACAAGCTACAGATGACTCTGGTCATGAATCCGACGGTTCCG CAGGCTCTGCTGCCGGCTCCGGCGAGTTTGCTGAAGCCGCTGCTAAAGAGGCTGCTGCTAAAG CCGGTTCTGCCGGTTCAGCAGCTGGATCTGGAGAATTTGGTTCCCCAAATGCTGGTAGTGTTGA ACAAACCCCAAAGAAACCTGGTTTGAGAAGAAGAGGTAGTGCTGGTTCTGCCGCTGGCTCCGGA GAATTTGGTTCAGCCGAAGCTGCGGCCAAAGAGGCTGCTGCAAAGGAGGCTGCGGCTAAGGAA GCCGCCGCTAAAGCCGGTTCAGCTGGTTCCGCGGCAGGCTCCGGGGAATTTGGTTCTTCTTATT ATCACCACCACCACCATCACTTGGAATCCACTAGTTTATACAAGAAAGCAGGCTCTGGTTCATTC GAAAACGTCACTCATGAATTCATTTTGGCAACCTTAGAAAACGAAAACGCTAAGTTGAGAAGATT AGAAGCAAAGTTGGAAAGAGAATTGGCTAGATTAAGAAATGAAGTAGCTTGGTTGGGTTCTGCG GGCTCGGCCGCTGGCTCTGGTGAATTCGCCGAAGCTGCGGCCAAGGAGGCTGCCGCAAAGGC CGGTTCTGCCGGTTCCGCAGCGGGATCCGGCGAATTTTCTTACTACCATCATCACCATCACCAC TTGGAATCCACAAGTTTATACAAGAAAGCGGGTTCTGGTTCACAAAAAGTTGAAGAATTGAAAAA TAAGATAGCAGAATTGGAAAACAGAAACGCTGTAAAGAAAAATAGAGTTGCACATTTGAAGCAAG AAATCGCTTACTTGAAGGATGAATTAGCAGCCCATGAATTCGAAGGTAGTGCCGGTTCCGCTGC TGGCTCAGGCGAATTTGGTAGTGCAGAAGCTGCCGCTAAGGAGGCTGCCGCCAAAGAAGCAGC CGCAAAAGAAGCTGCCGCAAAAGCCGGTTCTGCGGGCTCTGCTGCCGGATCCGGCGAATTCGG TTCAGTCTCCAGTACTAAATTAGTATCCTTTCATGATGACAGTGATGAAGACTTGTTACATATCGG TTCTGCAGGCTCAGCCGCTGGCTCTGGAGAGTTTGCAGAGGCAGCTGCTAAAGAAGCCGCCGC AAAGGCAGGTTCTGCAGGTTCTGCAGCTGGTAGTGGTGAATTCGCTGCTGCAACCCCAATATCT ACTTTTCATGATGACTCAGACGAAGACTTGTTGCATGTCGGTTCCGCAGGTTCAGCAGCGGGAT CCGGTGAATTTGGTTCAGCAGAAGCTGCCGCCAAGGAGGCCGCTGCTAAAGAAGCAGCAGCCA AGGAAGCAGCAGCAAAGGCCGGCTCTGCTGGTTCTGCTGCCGGGTCCGGCGAATTTGGTTCTT CTTATTACCACCATCATCATCACCACTTGGAATCCACTAGTTTATATAAGAAAGCCGGTTCTGGTT CACAAAAGGTGGAATCATTAAAACAAAAGATTGAAGAATTGAAGCAAAGAAAAGCACAATTGAAA AATGATATTGCCAATTTGGAAAAGGAAATCGCTTACGCAGAAACAGGTAGTGCCGGTTCAGCCG CGGGCTCTGGTGAATTCGCAGAAGCTGCCGCAAAAGAAGCTGCAGCAAAAGCCGGTTCTGCAG GCTCTGCTGCTGGCTCTGGCGAATTTTCCTACTATCATCATCATCATCATCACTTGGAATCCACA AGTTTATACAAGAAAGCGGGTAGTGAATTTTTCAGAAGAGAAAGAAACAAGATGGCAGCCGCTAA GTGTAGAAACAGAAGAAGAGAATTGACTGATACATTACAAGCTGAAACAGATCAATTAGAAGACG AAAAATCAGCTTTGCAAACCGAAATCGCAAATTTGTTGAAAGAAAAAGAAAAATTGGAATTCATTT TAGCAGCCCATAGACCAGCTTGCAAAATACCTGATGACTTGGGTTTTCCAGAAGAAATGTCTTTA GAAGGTAGTGCCGGTAGTGCCGCTGGCTCAGGTGAATTTGGTAGTGCAGAAGCTGCCGCGAAA GAAGCCGCAGCTAAAGAAGCTGCCGCCAAAGAGGCAGCCGCAAAGGCAGGTTCAGCAGGTTCA GCTGCCGGGTCCGGGGAATTTGGTTCATTCCAAATGCCAGCTGACACTCCTCCACCTGCATATT TGCCACCTGAAGATCCTATGACAGGTAGTGCCGGTTCTGCTGCCGGGTCTGGCGAATTCGCTGA AGCCGCTGCTAAGGAGGCTGCAGCTAAGGCCGGCTCTGCAGGTTCCGCTGCAGGTTCAGGTGA ATTTGAAAGAGAATCTAACGAAGAACCACCTCCACCTTATGAAGATCCATACTGGGGTAATGGTG GTTCTGCCGGTAGTGCCGCCGGCTCAGGCGAATTTGGTTCTGCGGAGGCTGCTGCAAAGGAAG CTGCGGCCAAGGAAGCTGCCGCAAAAGAGGCTGCTGCCAAGGCCGGTTCAGCAGGTTCAGCA GCTGGGTCCGGTGAATTTGGTTCCAGTTATTATCACCACCATCATCACCACTTGGAATCTACCTC ATTATATAAGAAAGCGGGTTCCGGTAGTCAAAAAGTTGCAGAATTGAAAAACAGAGTTGCTGTCA AATTAAATAGAAATGAGCAGTTGAAAAATAAGGTCGAGGAGTTGAAAAATAGAAACGCATACTTG AAAAATGAATTGGCTACTTTGGAAAACGAAGTCGCAAGATTAGAAAATGATGTAGCTGAAGGCTC TGCTGGTTCCGCAGCGGGCTCAGGTGAATTCGCCGAAGCAGCCGCAAAGGAAGCTGCCGCTAA

GGCCGGCTCAGCAGGTTCTGCCGCCGGAAGCGGTGAATTTTCTTATTACCACCACCACCATCAC CACTTGGAATCTACTTCATTATACAAGAAAGCGGGGTCCGGTAGTAACGAAGTCACAACCTTAGA AAATGATGCAGCCTTTATAGAAAACGAAAATGCCTACTTAGAAAAAGAAATTGCAAGATTGAGAA AGGAAAAAGCTGCATTGAGAAACAGATTAGCCCACAAGAAATCTTACTATCACCACCATCATCAT CACTTGGAATCTACATCATTATACAAGAAAGCGGGCTCCGGTAGTGCTAGAAATGCCTACTTAAG AAAGAAAATAGCCAGATTGAAGAAAGACAATTTGCAATTAGAGAGAGATGAACAGAACTTAGAAA AGATTATAGCCAATTTGAGAGATGAAATTGCTAGATTAGAAAATGAAGTAGCTTCTCATGAACAAG GTAGTGCTGGCTCCGCTGCCGGCTCCGGAGAATTTGCCGAAGCTGCCGCCAAGGAAGCCGCG GCCAAGGCTGGTTCCGCTGGTTCTGCTGCCGGATCTGGAGAATTTTCCTATTACCATCATCATCA TCATCATTTGGAATCTACATCATTATACAAGAAAGCGGGATCTGGTTCTAACTTGGTCGCCCAATT GGAGAACGAAGTCGCATCATTGGAGAACGAAAACGAAACCTTGAAGAAAAAGAACTTACACAAA AAGGATTTGATAGCTTACTTAGAAAAAGAAATCGCTAATTTGAGAAAGAAAATTGAAGAAGGTAGT GCAGGTTCAGCCGCTGGCTCCGGTGAATTTGGTTCAGCGGAGGCTGCCGCTAAGGAGGCAGCC GCTAAAGAAGCAGCCGCTAAGGAGGCTGCAGCAAAAGCAGGTTCCGCAGGTTCTGCAGCGGGT TCCGGAGAATTTGGTTCTGAACAAAAGTTGATCTCTGAAGAAGATTTGGAACAAAAGTTGATATC TGAAGAAGACTTGGAACAAAAATTAATATCAGAAGAAGATTTGGGTAGTGCAGGTTCAGCAGCTG GTTCTGGAGAATTTGGTTCAGCAGGTTCTGCCGCTGGAAGTGGCGAATTCGGTAGTGCCGGCTC CGCTGCTGGCTCTGGCGAATTTGGTTCTGGT

2. Malonyl-CoA Metabolon Scaffold (MCASCFLD) - (FLAGh

ATGGGTTCTGCTGGTTCAGCTGCAGGTTCTGGTGAATTTGGTTCCGCAGGTAGTGCCGCTGGTT CTGGTGAATTCGGTTCTGCTGGTTCAGCAGCCGGTTCTGGTGAATTTTCATATTACCATCACCAT CACCATCACTTGGAATCCACCAGTTTATACAAAAAGGCTGGTTCTGGTTCAGCTAGAAACGCATA TTTGAGAAAGAAAATTGCTAGATTGAAGAAAGATAACTTGCAATTGGAAAGAGATGAACAAAATTT GGAAAAGATTATCGCCAACTTAAGAGATGAAATAGCAAGATTGGAAAACGAAGTAGCTTCTCATG AACAAGGTTCCGCAGGTAGTGCAGCTGGTTCTGGTGAATTTGCTGAAGCCGCTGCAAAGGAAGC CGCTGCAAAAGCTGGTTCCGCTGGTTCAGCCGCTGGTTCCGGTGAATTCAGTTACTATCACCAT CACCATCATCACTTGGAATCCACTAGTTTATATAAAAAGGCCGGTTCTGGTTCAAATTTGGTTGCT CAATTAGAAAACGAAGTCGCATCTTTAGAAAACGAAAACGAAACATTGAAAAAGAAAAATTTGCAT AAGAAAGATTTGATCGCTTATTTGGAAAAGGAAATCGCAAACTTGAGAAAGAAAATAGAAGAAGG TTCCGCTGGTTCTGCTGCTGGTTCCGGTGAATTTGGTTCAGCTGAAGCTGCAGCCAAGGAAGCT GCAGCCAAAGAAGCCGCTGCTAAAGAAGCTGCAGCCAAAGCAGGTTCTGCCGGTTCTGCCGCA GGTTCCGGTGAATTCGGTTCTTCAGCTACTAGAGAATTGGATGAATTGATGGCATCCTTAAGTGA CTTCAAGATACAAGGTGGTTCCGCTGGTTCTGCAGCCGGCTCTGGCGAATTCGCAGAAGCAGC CGCTAAGGAAGCAGCCGCTAAAGCTGGTTCTGCAGGTTCTGCTGCCGGTTCTGGTGAATTCGAT TTGGCTTTGTCTGAAAACTGGGCACAAGAATTCTTGGCTGCAGGTGACGCTGTTGATGGTTCTG CTGGTAGTGCTGCCGGTTCAGGTGAATTTGGTAGTGCTGAAGCTGCTGCCAAAGAAGCAGCCG CTAAAGAAGCTGCTGCCAAGGAAGCTGCCGCTAAAGCAGGTTCCGCCGGTTCTGCCGCCGGCT CCGGCGAATTTGGTTCAGATTATAAGGATGACGATGACAAGGATTACAAAGACGATGATGACAA GGATTATAAAGATGACGATGACAAAGGTTCCGCTGGTAGTGCCGCCGGCTCTGGAGAATTCGGT TCTGCCGGTTCAGCTGCCGGCTCCGGAGAATTTGGTTCCGCTGGTAGTGCAGCCGGTTCAGGT GAATTCGGTTCTGGT

FIG 15A

Complete HCA Gene Cassette Nucleotide Sequence

ATGAGTGCTAAGGCAATTTCTGAACAAACTGGTAAAGAATTGTTGTACAAGTTTATTTGTACTACA TCAGCCATCCAAAATAGATTCAAATACGCTAGAGTTACCCCAGATACTGACTGGGCTAGATTGTT ACAAGATCATCCATGGTTGTTATCTCAAAACTTGGTTGTCAAACCTGACCAATTAATTAAGAGAAG AGGTAAATTGGGTTTAGTAGGTGTTAATTTGACATTGGATGGTGTAAAGTCTTGGTTGAAACCAA GATTAGGTCAAGAAGCCACAGTTGGTAAAGCTACCGGTTTCTTGAAAAATTTCTTGATCGAACCA TTTGTCCCTCATTCACAAGCCGAAGAATTCTATGTATGTATCTACGCTACTAGAGAGGGTGACTA TGTTTTATTTCATCACGAAGGTGGTGTCGACGTAGGTGACGTTGACGCCAAGGCTCAAAAGTTGT TGGTTGGTGTCGATGAAAAGTTGAACCCAGAAGACATTAAAAAGCATTTGTTGGTTCACGCACCT GAAGATAAAAAGGAAATATTGGCCTCCTTTATAAGTGGTTTGTTTAATTTCTACGAAGATTTGTAC TTCACCTACTTGGAAATTAACCCATTAGTAGTTACTAAGGATGGTGTATATGTTTTGGACTTAGCT GCAAAAGTTGATGCAACAGCCGACTACATTTGTAAGGTCAAATGGGGTGACATCGAATTTCCACC TCCATTCGGTAGAGAAGCTTATCCAGAAGAAGCCTACATTGCTGATTTGGACGCTAAGTCTGGTG CATCATTGAAGTTGACATTGTTGAACCCTAAAGGTAGAATTTGGACCATGGTTGCTGGTGGTGGT GCTAGTGTCGTATATTCTGATACTATATGCGACTTGGGTGGTGTTAACGAATTGGCAAACTACGG TGAATACTCAGGTGCCCCATCCGAACAACAAACATACGATTACGCTAAGACCATCTTGTCCTTAA TGACTAGAGAAAAGCATCCTGATGGTAAAATCTTGATCATCGGTGGTAGTATCGCAAATTTTACT AACGTTGCCGCTACATTCAAGGGTATCGTCAGAGCTATAAGAGATTACCAAGGTCCATTGAAGG AACACGAAGTAACAATATTCGTTAGAAGAGGTGGTCCTAACTACCAAGAAGGTTTGAGAGTCATG GGTGAAGTAGGTAAAACCACTGGTATACCAATCCATGTCTTTGGTACAGAAACCCACATGACTGC AATAGTTGGTATGGCCTTAGGTCATAGACCAATCCCTAATCAACCTCCAACCGCAGCCCACACTG CAAATTTCTTGTTAAACGCCTCTGGTTCAACTTCCACACCAGCTCCTTCTAGAACAGCAAGTTTCT CTGAATCAAGAGCTGATGAAGTCGCTCCAGCTAAGAAAGCAAAACCAGCCATGCCTCAAGACTC CGTTCCAAGTCCTAGATCTTTGCAGGGTAAATCTACTACTTTGTTTTCTAGACATACTAAGGCTAT AGTATGGGGTATGCAAACAAGAGCAGTTCAAGGCATGTTGGATTTCGACTATGTTTGTAGTAGAG ATGAACCATCTGTTGCTGCAATGGTCTATCCTTTTACTGGTGACCATAAGCAAAAATTCTACTGG GGTCACAAGGAAATATTGATCCCAGTTTTTAAGAACATGGCCGATGCTATGAGAAAACATCCTGA AGTCGACGTATTGATTAACTTCGCCTCATTAAGATCCGCTTACGATTCTACAATGGAAACCATGA ACTACGCTCAAATAAGAACCATCGCTATCATTGCAGAAGGTATTCCAGAAGCCTTGACTAGAAAG TTGATTAAGAAAGCTGATCAAAAAGGTGTCACAATAATCGGTCCAGCTACCGTAGGTGGTATTAA GCCTGGTTGTTTCAAGATCGGTAACACTGGTGGTATGTTGGATAACATATTGGCATCTAAGTTGT ATAGACCAGGTTCAGTCGCTTACGTATCCAGAAGTGGTGGTATGTCCAACGAATTGAACAACATC ATCAGTAGAACTACAGATGGTGTATACGAAGGTGTTGCTATTGGTGGTGACAGATACCCAGGTT CTACTTTTATGGATCATGTATTGAGATATCAAGACACACCTGGTGTTAAAATGATTGTTGTCTTGG GTGAAATAGGTGGTACTGAAGAATACAAGATATGCAGAGGTATCAAAGAAGGTAGATTGACAAA GCCAATCGTTTGTTGGTGCATTGGTACTTGTGCAACAATGTTTTCTTCAGAAGTTCAATTCGGTCA TGCAGGTGCCTGCGCTAATCAAGCTTCAGAAACAGCAGTTGCCAAGAACCAAGCATTAAAAGAA GCCGGTGTTTTTGTCCCTAGATCTTTCGATGAATTAGGTGAAATCATTCAATCAGTCTATGAAGAC TTGGTAGCTAATGGTGTAATTGTTCCAGCACAAGAAGTTCCTCCACCTACTGTCCCTATGGATTA CTCTTGGGCAAGAGAATTGGGTTTAATTAGAAAGCCAGCTAGTTTTATGACCTCTATATGTGATG AAAGAGGTCAAGAATTGATCTATGCTGGTATGCCTATTACTGAAGTATTCAAAGAAGAAATGGGT ATCGGTGGTGTTTTAGGTTTGTTGTGGTTCCAAAAGAGATTGCCAAAGTACTCTTGTCAATTCATT GAAATGTGCTTAATGGTTACAGCTGATCATGGTCCTGCTGTCTCAGGTGCACACAATACCATAAT CTGCGCTAGAGCTGGTAAAGATTTGGTTTCTTCTTTGACCTCAGGTTTGTTAACTATTGGTGACA GATTTGGTGGTGCATTAGACGCCGCTGCAAAGATGTTTTCAAAAGCTTTCGATTCCGGTATAATC CCAATGGAATTCGTTAATAAGATGAAAAAGGAGGGTAAATTGATAATGGGTATCGGTCATCGTGT TAAGTCTATCAATAACCCTGATATGAGAGTACAAATCTTGAAGGACTATGTTAGACAACACTTTCC AGCCACACCTTTGTTAGATTACGCTTTGGAAGTTGAAAAGATTACCACTTCTAAAAAGCCAAATTT GATCTTGAACGTTGATGGTTTAATTGGTGTTGCTTTTGTCGACATGTTGAGAAACTGTGGTTCCTT CACTAGAGAAGAAGCTGATGAATATATCGACATTGGTGCATTGAATGGTATCTTTGTTTTAGGTA

GATCTATGGGTTTCATTGGTCATTACTTGGATCAAAAGAGATTAAAGCAAGGTTTGTACAGACAT CCATGGGATGACATTTCTTACGTTTTACCTGAACACATGTCAATGAAATTGTCTGGTGGTGGTGG TTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCTTGGTACAATTTGGGTAACGCA TACTACAAGCAGGGTGACTACCAAAAGGCAATTGAATATTACCAAAAGGCCTTGGAATTAGACCC AAATAACGCAGAAGCCTGGTATAATTTGGGTAATGCTTATTATAAACAGGGTGACTATCAAAAGG CTATCGAATACTACCAAAAGGCATTGGAATTAGACCCTAATAACGCTGAAGCATGGTATAATTTG GGTAACGCTTATTATAAGCAGGGTGACTATCAAAAAGCCATCGAAGACTACCAAAAGGCTTTGGA ATTAGATCCAAATAACTTACAAGCCGAAGCTTGGAAGAATTTGGGTAACGCTTACTATAAACAGG GTGACTACCAAAAAGCAATTGAATACTATCAAAAAGCTTTAGAATTGGACCCTAATAACGCATCA GCCTGGTACAATTTGGGTAATGCTTACTATAAGCAGGGTGACTATCAGAAGGCCATTGAATACTA TCAAAAGGCTTTAGAATTGGATCCAAATAACGCTAAAGCATGGTACAGACGTGGTAACGCTTATT ACAAACAGGGTGACTACCAGAAAGCCATTGAAGATTATCAAAAGGCTTTGGAATTGGATCCTAAC AACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTT CTTCATATTACCATCACCATCACCATCACTTGGAATCCACAAGTTTATACAAAAAGGCTGGTTCTG GTTCAAATTTGGTCGCACAATTGGAAAACGAAGTAGCCTCTTTAGAAAATGAAAACGAAACCTTG AAAAAGAAAAACTTACATAAGAAAGATTTGATCGCTTATTTGGAAAAGGAAATCGCAAATTTGAGA AAGAAAATTGAAGAAGGTAGTGCAGGTTCTGCCGCTGGTTCTGGTGAATTTGGTTCAGCTGAAG CAGCCGCTAAGGAAGCAGCCGCTAAAGCCGGTTCAGCTGGTTCCGCAGCCGGTTCTGGTGAAT TCGGTTCCAGTTACTATCACCATCACCATCATCACTTGGAATCCacaAGTTTATATAAGAAAGCAG GTTCTGGTTCAGCAAGAAATGCCTACTTGAGAAAGAAAATAGCTAGATTAAAGAAAGATAACTTG CAATTGGAAAGAGATGAACAAAATTTGGAAAAGATTATCGCCAACTTAAGAGATGAAATCGCTAG ATTGGAAAATGAAGTTGCATCCCATGAACAAGGTAGTGGTGCTACTAACTTCTCTTTGTTGAAGC AAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGAAAAACTGTGTAATCGTTTCTGCTGTTAGA ACTGCAATTGGTTCCTTTAATGGTAGTTTGGCCTCTACATCAGCTATTGATTTGGGTGCTACCGT CATCAAAGCTGCAATTGAAAGAGCAAAGATTGATTCTCAACATGTCGACGAAGTAATAATGGGTA ACGTTTTGCAAGCTGGTTTAGGTCAAAATCCAGCAAGACAAGCCTTGTTAAAATCTGGTTTAGCA GAAACTGTATGTGGTTTCACAGTTAATAAGGTCTGCGGTTCTGGTTTGAAGTCAGTTGCTTTAGC CGCTCAAGCTATACAAGCAGGTCAAGCCCAATCTATCGTCGCTGGTGGTATGGAAAATATGTCAT TGGCACCTTATTTGTTAGATGCAAAAGCCAGATCAGGTTATAGATTAGGTGACGGTCAAGTATAC GACGTTATTTTGAGAGATGGTTTAATGTGCGCTACTCATGGTTATCACATGGGTATTACAGCAGA AAATGTTGCCAAAGAATACGGTATAACCAGAGAAATGCAAGATGAATTGGCATTACATTCCCAAA GAAAGGCAGCCGCTGCAATCGAAAGTGGTGCTTTTACTGCAGAAATTGTCCCAGTAAACGTTGT CACAAGAAAGAAAACTTTCGTTTTCTCCCAAGATGAATTCCCAAAAGCTAATAGTACCGCTGAAG CATTGGGTGCTTTAAGACCTGCATTCGACAAGGCCGGTACCGTAACTGCCGGTAATGCTTCTGG TATAAACGATGGTGCCGCTGCATTGGTTATCATGGAAGAATCAGCCGCTTTAGCAGCCGGTTTG ACACCTTTAGCTAGAATTAAATCTTATGCATCAGGTGGTGTTCCACCTGCTTTGATGGGTATGGG TCCAGTCCCTGCTACCCAAAAGGCATTGCAATTAGCCGGTTTGCAATTGGCTGATATCGACTTAA TCGAAGCAAACGAAGCCTTTGCTGCACAATTCTTGGCAGTTGGTAAAAATTTGGGTTTCGACTCC GAAAAGGTTAATGTCAACGGTGGTGCCATTGCTTTGGGTCATCCAATAGGTGCTTCAGGTGCAA GAATCTTGGTTACATTGTTGCATGCCATGCAAGCTAGAGATAAAACCTTGGGTTTAGCTACTTTGT GTATCGGTGGTGGTCAAGGTATCGCAATGGTTATCGAAAGATTGAATAAGTTGTCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTACAATTTGGGTAAC GCTTACTACAAGCAGGGTGACTACCAAAAGGCAATCGAATACTACCAAAAGGCCTTGGAATTAG ATCCAAATAACGCTGAAGCATGGTATAATTTGGGTAATGCCTATTATAAACAGGGTGACTATCAA AAAGCTATTGAATATTACCAAAAGGCATTGGAATTAGATCCTAATAACGCCGAAGCTTGGTATAAT TTGGGTAACGCCTATTATAAGCAGGGTGACTATCAAAAGGCCATCGAAGATTACCAAAAGGCTTT GGAATTGGATCCAAACAACTTGCAAGCAGAAGCCTGGAAGAATTTGGGTAACGCTTATTACAAAC AGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAAGCCTTAGAATTGGATCCTAATAACGCTT CTGCATGGTACAATTTGGGTAATGCCTACTATAAACAGGGTGACTACCAGAAGGCTATTGAATAC TACCAAAAAGCATTAGAATTGGATCCAAATAACGCCAAGGCTTGGTACAGACGTGGTAATGCCTA TTACAAGCAGGGTGACTACCAGAAAGCCATAGAAGACTATCAAAAAGCCTTGGAATTGGATCCTA ACAACAGATCCAGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTG

CTTCTTCATATTACCATCACCATCACCATCACTTGGAATCTACATCATTATACAAAAAGGCTGGTT CCGGTAGTAATGAAGTTACTACATTGGAAAACGATGCCGCTTTTATCGAAAACGAAAACGCATAC TTGGAAAAGGAAATCGCCAGATTAAGAAAGGAAAAGGCAGCCTTGAGAAATAGATTAGCCCATA AAAAGGGTTCCGCTGGTAGTGCTGCAGGTTCTGGTGAATTTGGTTCAGCTGAAGCCGCTGCAAA AGAAGCCGCTGCAAAGGCAGGTTCTGCCGGTTCAGCCGCTGGTTCTGGTGAATTCGGTTCCAGT TACTATCACCATCACCATCATCACTTGGAATCTACTTCATTATATAAAAAGGCCGGTTCCGGTAGT CAAAAAGTCGCTGAATTAAAGAACAGAGTAGCTGTTAAGTTGAACAGAAACGAACAATTGAAAAA TAAGGTAGAAGAATTGAAAAATAGAAACGCCTACTTAAAGAATGAATTGGCAACATTGGAAAACG AAGTCGCTAGATTGGAAAATGATGTAGCAGAAGGTTCTGGTGCTACTAACTTCTCTTTGTTGAAG CAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGAAAAAGGTTTGTGTCATTGGTGCTGGTA CCATGGGTTCTGGTATAGCACAAGCCTTTGCTGCAAAAGGTTTCGAAGTTGTCTTGAGAGATATC AAGGACGAATTCGTTGATAGAGGTTTGGACTTCATCAATAAGAACTTGTCTAAGTTGGTTAAAAA GGGTAAAATCGAAGAAGCTACAAAGGTAGAAATCTTGACCAGAATTTCAGGTACTGTTGATTTGA ATATGGCCGCTGATTGTGACTTGGTAATCGAAGCAGCCGTTGAAAGAATGGATATTAAGAAACAA ATCTTCGCAGATTTGGACAACATCTGCAAACCTGAAACAATCTTAGCCTCAAACACCTCTTCATTG TCCATTACTGAAGTCGCTAGTGCAACAAAAAGACCAGATAAGGTAATAGGCATGCATTTCTTTAA TCCAGCTCCTGTTATGAAGTTGGTAGAAGTTATTAGAGGTATAGCAACATCTCAAGAAACCTTTG ACGCTGTTAAGGAAACTTCAATAGCAATCGGTAAAGATCCAGTCGAAGTAGCCGAAGCTCCTGG TTTCGTAGTTAACAGAATCTTGATACCTATGATCAACGAAGCTGTTGGTATCTTGGCTGAAGGTAT TGCATCTGTCGAAGATATTGACAAAGCCATGAAGTTAGGTGCTAATCACCCAATGGGTCCTTTGG AATTGGGTGACTTTATTGGTTTGGACATATGTTTAGCTATCATGGACGTTTTGTATTCCGAAACAG GTGACAGTAAATACAGACCACATACCTTGTTGAAGAAATATGTTAGAGCAGGTTGGTTAGGTAGA AAGTCTGGTAAAGGTTTCTACGATTACTCTAAAAAGTTGTCTGGTGGTGGTGGTTCTGGTGGTGG TGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTACAATTTGGGTAACGCTTACTACAAGCAG GGTGACTACCAAAAGGCCATAGAATACTACCAAAAGGCTTTGGAATTGGATCCTAATAACGCTGA AGCATGGTATAATTTGGGTAATGCATATTATAAACAGGGTGACTATCAAAAGGCAATCGAATACT ACCAAAAGGCCTTGGAATTAGATCCAAATAACGCCGAAGCTTGGTATAATTTGGGTAACGCCTAT TATAAGCAGGGTGACTATCAAAAAGCTATCGAAGATTACCAAAAGGCATTGGAATTGGATCCTAA CAACTTACAAGCAGAAGCCTGGAAGAATTTGGGTAACGCATATTACAAACAGGGTGACTACCAAA AAGCCATTGAATATTATCAAAAAGCTTTGGAATTGGATCCAAATAACGCTTCAGCATGGTACAATT TGGGTAATGCCTATTACAAGCAGGGTGACTATCAGAAAGCTATTGAATATTATCAAAAGGCTTTG GAATTAGATCCTAATAACGCCAAGGCTTGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTG ACTACCAGAAGGCCATTGAAGACTATCAAAAAGCCTTGGAATTGGATCCAAACAACAGATCTAGA TCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCCGAAAATTTGT ACTTCCAAGGTGAAAACTTGTACTTCCAGGGTGACTCCAGTGAAAGTTGTTGGAATTGCGGTAGA AAAGCCTCCGAAACCTGTAGTGGTTGCAACACTGCTAGATATTGTGGTTCTTTTTGCCAACACAA AGATTGGGAAAAGCATCACCATATTTGTGGTCAAACATTACAAGCACAACAAGGTTCTGCCGGTT CAGCTGCAGGTTCTGGTGAATTTGGTTCCGCTGAAGCCGCTGCAAAAGAAGCCGCTGCAAAGG CAGGTTCCGCCGGTAGTGCCGCTGGTAGTGGTGAATTCGGTTCTATGGCAGTTTCCGAAAGTCA ATTGAAGAAAATGGTTTCTAAGTACAAGTACAGAGATTTGACTGTTAGAGAAACAGTTAACGTCAT CACTTTGTACAAGGATTTGAAGCCAGTCTTGGACTCATACGTTTTTAATGATGGTTCTTCAAGAGA ATTGATGAACTTAACTGGTACAATACCAGTTCCTTACCGTGGTAACACTTACAACATCCCAATCTG TTTGTGGTTGTTAGATACATATCCTTACAATCCACCTATCTGCTTCGTCAAACCAACATCCAGTAT GACCATTAAAACTGGTAAACATGTTGATGCTAACGGTAAAATATATTTGCCATACTTACACGAATG GAAGCATCCTCAATCAGACTTGTTGGGTTTAATCCAAGTAATGATCGTCGTATTTGGTGACGAAC CACCTGTTTTCTCTAGACCAGGTTCAGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCAGGTGAC GTTGAAGAAAATCCAGGTCCAATGGAATTGAACAACGTTATATTGGAAAAGGAGGGTAAAGTCG CTGTTGTCACTATAAATAGACCAAAGGCATTGAACGCCTTGAACTCTGATACATTGAAGGAAATG GACTACGTTATCGGTGAAATTGAAAACGATTCAGAAGTCTTAGCAGTAATTTTGACCGGTGCCGG TGAAAAATCCTTTGTTGCCGGTGCTGATATCAGTGAAATGAAGGAAATGAACACTATCGAAGGTA GAAAGTTCGGTATCTTGGGTAACAAGGTTTTCAGAAGATTGGAATTGTTGGAAAAGCCTGTTATA GCTGCAGTCAATGGTTTCGCTTTGGGTGGTGGTTGTGAAATCGCAATGTCCTGCGATATTAGAAT

AGCTTCTTCAAACGCAAGATTTGGTCAACCAGAAGTCGGTTTAGGTATTACACCTGGTTTCGGTG GTACCCAAAGATTATCTAGATTGGTTGGTATGGGTATGGCCAAGCAATTGATTTTTACTGCTCAA AACATCAAGGCTGATGAAGCATTGAGAATCGGTTTGGTTAATAAGGTAGTTGAACCATCTGAATT GATGAACACCGCCAAGGAAATCGCTAATAAGATTGTTTCTAATGCTCCAGTTGCTGTCAAGTTGA GTAAGCAAGCTATAAATCGTGGTATGCAATGTGATATCGACACTGCATTGGCCTTCGAATCTGAA GCATTTGGTGAATGCTTCTCAACAGAAGATCAAAAAGACGCAATGACCGCCTTTATCGAAAAGAG AAAGATAGAAGGTTTCAAAAACAGAAAGTTATCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCT GGTGGTGGTGGTAGTGCTGAAGCATGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACT ACCAAAAGGCAATCGAATACTACCAAAAGGCCTTGGAATTGGACCCAAATAACGCCGAAGCTTG GTATAATTTGGGTAATGCCTATTATAAACAGGGTGACTATCAAAAAGCTATAGAATACTACCAAAA GGCATTGGAATTGGACCCTAATAACGCAGAAGCCTGGTATAATTTGGGTAACGCCTATTATAAGC AGGGTGACTATCAAAAGGCCATAGAAGACTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTA CAAGCTGAAGCATGGAAGAATTTGGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTAT TGAATATTATCAAAAAGCTTTAGAATTAGACCCTAATAACGCCTCTGCTTGGTACAATTTGGGTAA TGCCTACTATAAACAGGGTGACTACCAGAAGGCTATTGAATATTACCAAAAAGCTTTAGAATTGG ATCCAAATAACGCAAAGGCCTGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTACCA GAAAGCCATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCTAACAACAGATCCAGAAGTGCTG GTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTGGTCCATTGGGTTCCC CTTTGACTGCATCAATGTTAGCTTCCGCACCACCTCAAGAACAAAAGCAAATGTTGGGTGAAAGA TTATTCCCATTGATACAAGCTATGCATCCTACTTTAGCAGGTAAAATCACAGGCATGTTGTTGGAA ATCGATAACTCTGAATTGTTACACATGTTAGAATCCCCAGAAAGTTTGAGATCTAAAGTTGACGAA GCCGTAGCTGTTTTGCAAGCTCATCAAGCAAAAGAAGCCGCTCAAAAGGCCGGTTCAGCTGGTT CCGCAGCCGGTAGTGGTGAATTTGGTTCTGCTGAAGCTGCAGCCAAAGAAGCTGCAGCCAAGG CAGGTAGTGCCGGTTCTGCTGCAGGTTCTGGTGAATTCGGTTCCAATACCAACATGAGTGTCCC AACTGATGGTGCTGTAACTACATCTCAAATTCCTGCATCAGAACAAGAAACTTTAGTTAGACCAAA GCCTTTGTTGTTGAAGTTGTTGAAGTCAGTAGGTGCTCAAAAAGATACCTACACTATGAAGGAAG TTTTATTTTATTTGGGTCAATACATCATGACAAAGAGATTATACGATGAAAAGCAACAACATATCG TTTACTGTTCAAACGATTTGTTGGGTGACTTGTTTGGTGTACCATCTTTCTCAGTTAAGGAACACA GAAAGATCTATACAATGATATACAGAAATTTGGTCGTAGGTTCTGGTGCTACTAACTTCTCTTTGT TGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGATCGTAAAGCCAATGGTTAGAAA CAACATCTGTTTGAACGCTCATCCTCAAGGTTGCAAAAAGGGTGTAGAAGATCAAATCGAATACA CCAAAAAGAGAATCACTGCAGAAGTTAAAGCCGGTGCTAAAGCACCTAAGAATGTTTTGGTCTTA GGTTGTTCCAACGGTTATGGTTTGGCTAGTAGAATAACAGCTGCATTTGGTTACGGTGCCGCTAC CATCGGTGTTTCCTTCGAAAAGGCTGGTAGTGAAACCAAATATGGTACTCCAGGTTGGTACAATA ACTTGGCATTTGATGAAGCAGCCAAGAGAGAAGGTTTATACTCTGTCACTATAGATGGTGACGCT TTCTCAGATGAAATCAAGGCACAAGTTATTGAAGAAGCCAAAAAGAAAGGTATAAAATTCGATTT GATCGTTTACTCCTTAGCAAGTCCAGTCAGAACAGATCCTGACACCGGTATAATGCATAAGTCTG TTTTGAAGCCATTCGGTAAAACTTTCACAGGTAAAACAGTCGATCCTTTCACCGGTGAATTGAAA GAAATATCTGCTGAACCAGCAAATGATGAAGAAGCTGCAGCCACAGTAAAAGTTATGGGTGGTG AAGACTGGGAAAGATGGATCAAGCAATTGTCCAAAGAAGGTTTGTTGGAAGAAGGTTGTATCAC CTTAGCTTATTCATACATTGGTCCTGAAGCCACTCAAGCTTTGTATAGAAAAGGTACAATCGGTAA AGCTAAAGAACATTTGGAAGCCACCGCTCACAGATTAAATAAGGAAAACCCATCTATCAGAGCAT TTGTTTCTGTAAATAAGGGTTTAGTTACTAGAGCATCCGCCGTTATCCCAGTCATTCCTTTGTATT TGGCTAGTTTGTTTAAGGTTATGAAGGAAAAGGGTAACCATGAAGGTTGCATAGAACAAATCACT AGATTGTACGCAGAAAGATTATACAGAAAGGATGGTACAATTCCAGTTGACGAAGAAAACAGAAT CAGAATCGATGACTGGGAATTGGAAGAAGATGTCCAAAAGGCAGTATCTGCCTTAATGGAAAAA GTTACCGGTGAAAACGCTGAATCATTGACTGATTTGGCAGGTTATAGACACGACTTTTTAGCCTC TAATGGTTTCGATGTCGAAGGTATTAACTACGAAGCAGAAGTAGAAAGATTCGACAGAATTAAAT TGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGT ATAATTTGGGTAACGCTTATTACAAGCAGGGTGACTACCAAAAGGCCATCGAATACTACCAAAAG GCTTTGGAATTGGACCCTAATAACGCCGAAGCTTGGTACAATTTGGGTAATGCCTACTATAAACA GGGTGACTATCAAAAAGCAATTGAATATTACCAAAAGGCCTTGGAATTAGACCCAAATAACGCAG

AAGCCTGGTACAATTTGGGTAACGCCTACTATAAGCAGGGTGACTATCAAAAGGCTATTGAAGAC TACCAAAAGGCATTGGAATTAGATCCTAATAACTTGCAAGCTGAAGCATGGAAAAATTTGGGTAA TGCCTATTATAAACAGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAAGCTTTGGAATTGGA CCCAAATAACGCCTCAGCTTGGTATAATTTGGGTAATGCATACTACAAACAGGGTGACTATCAGA AGGCAATAGAATACTATCAAAAAGCCTTAGAATTGGATCCTAATAACGCAAAAGCCTGGTATAGA CGTGGTAATGCCTACTACAAGCAGGGTGACTATCAGAAGGCGATAGAAGATTATCAAAAGGCAT TGGAATTGGATCCAAACAACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTC TGGTGGTGGTGGTGCTTCTTCATATTACCATCACCATCACCATCACTTGGAATCCACAAGTTTATA TAAGAAAGCAGGTTCTGGTTCAAATTTGTTAGCCACTTTGAGATCAACAGCTGCAGTATTGGAAA ACGAAAACCATGTTTTGGAAAAAGAAAAGGAAAAGTTGAGAAAGGAAAAGGAACAATTGTTGAAT AAGTTGGAAGCCTACAAAGGTTCTGCTGGTTCAGCCGCTGGTTCCGGTGAATTCGGTAGTGCTG AAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCTGGTTCCGCAGGTAGTGCAGCCGGTTCTGGTG AATTTGGTTCCAGTTACTATCACCATCACCATCATCACTTGGAATCCACAAGTTTATATAAGAAAG CTGGTTCTGGTTCAAAGAGAATCGCATACTTGAGAAAGAAAATCGCTGCATTAAAGAAAGATAAC GCCAACTTGGAAAAGGACATCGCTAATTTGGAAAACGAAATCGAAAGATTGATTAAAGAAATTAA AACATTAGAAAATGAAGTTGCTTCTCATGAACAAGGTTCAGGTGCTACTAACTTCTCTTTGTTGAA GCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGACTAGAGAAGTTGTCGTAGTTAGTGGT GTTAGAACAGCTATTGGTACCTTTGGTGGTTCTTTAAAAGATGTTGCACCAGCCGAATTGGGTGC ATTAGTCGTAAGAGAAGCTTTGGCAAGAGCCCAAGTTTCAGGTGACGATGTCGGTCATGTTGTC TTCGGTAACGTTATCCAAACAGAACCAAGAGATATGTATTTGGGTAGAGTAGCTGCAGTTAATGG TGGTGTTACCATAAACGCTCCTGCATTAACTGTCAACAGATTGTGTGGTAGTGGTTTACAAGCTA TTGTTTCTGCCGCTCAAACAATATTGTTAGGTGACACCGACGTTGCTATCGGTGGTGGTGCTGAA TCTATGTCAAGAGCCCCATACTTAGCTCCTGCAGCCAGATGGGGTGCCAGAATGGGTGACGCTG GTTTGGTTGACATGATGTTGGGTGCTTTGCATGATCCATTCCATAGAATCCACATGGGTGTAACT GCAGAAAACGTTGCCAAGGAATACGATATCTCAAGAGCACAACAAGACGAAGCTGCATTAGAAT CACACAGAAGAGCATCCGCCGCTATTAAAGCCGGTTACTTTAAGGATCAAATAGTTCCAGTAGTT TCTAAAGGTAGAAAGGGTGACGTTACCTTCGATACTGACGAACATGTTAGACACGACGCTACTAT TGATGACATGACAAAGTTAAGACCTGTTTTCGTCAAGGAAAATGGTACTGTTACAGCTGGTAATG CATCTGGTTTGAACGATGCAGCCGCTGCAGTCGTAATGATGGAAAGAGCCGAAGCTGAAAGAAG AGGTTTGAAACCATTAGCTAGATTGGTTTCTTATGGTCATGCTGGTGTCGATCCTAAAGCAATGG GTATAGGTCCAGTTCCTGCTACTAAGATCGCATTGGAAAGAGCCGGTTTACAAGTCTCTGATTTG GACGTAATTGAAGCCAATGAAGCTTTTGCCGCTCAAGCATGTGCCGTTACAAAAGCCTTGGGTTT AGATCCAGCTAAGGTCAATCCTAACGGTAGTGGTATCTCTTTAGGTCATCCAATTGGTGCAACCG GTGCCTTGATAACTGTTAAGGCTTTGCACGAATTGAACAGAGTACAAGGTAGATATGCATTAGTT ACAATGTGCATCGGTGGTGGTCAAGGTATTGCAGCCATATTCGAAAGAATTAAGTTGTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTACAATTTGG GTAACGCTTACTACAAGCAGGGTGACTACCAAAAGGCAATCGAATATTACCAAAAAGCCTTGGAA TTAGACCCAAATAACGCCGAAGCTTGGTATAATTTGGGTAATGCCTATTATAAACAGGGTGACTA TCAAAAAGCTATAGAATACTACCAAAAGGCATTGGAATTAGACCCTAATAACGCAGAAGCCTGGT ATAATTTGGGTAACGCCTATTATAAGCAGGGTGACTATCAAAAGGCCATAGAAGACTACCAAAAG GCTTTGGAATTGGATCCAAACAACTTACAAGCTGAAGCATGGAAGAATTTGGGTAACGCTTATTA CAAACAGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAGGCTTTAGAATTGGACCCTAATA ACGCCTCTGCTTGGTACAATTTGGGTAATGCCTACTATAAACAGGGTGACTACCAGAAGGCTATC GAATATTATCAAAAAGCTTTAGAATTGGACCCAAATAACGCAAAGGCCTGGTACAGACGTGGTAA TGCCTATTACAAGCAGGGTGACTACCAGAAAGCTATTGAAGATTATCAAAAGGCATTGGAATTGG ATCCTAACAACAGATCCAGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGG TGGTGCTTCTGATGTTATGTGGGAATATAAGTGGGAAAATACAGGTGACGCTGAATTATACGGTC CTTTTACTTCAGCACAAATGCAAACATGGGTATCCGAAGGTTATTTCCCTGATGGTGTTTACTGCA GAAAATTAGACCCACCTGGTGGTCAATTCTACAACTCAAAGAGAATAGATTTCGACTTGTACACC GGTTCAGCTGGTTCCGCTGCAGGTTCTGGTGAATTTGGTTCCGCAGAAGCCGCTGCAAAAGAAG CCGCTGCAAAGGCTGGTAGTGCAGGTTCTGCCGCTGGTAGTGGTGAATTTGGTTCTGAATCAGA TTCCGTCGAATTCAATAACGCTATATCTTACGTAAATAAGATTAAAACCAGATTTTTAGATCATCCA

GAAATCTATAGATCATTCTTAGAAATCTTGCATACATACCAAAAAGAACAATTGCACACCAAGGGT AGACCTTTCAGAGGCATGTCCGAAGAAGAAGTCTTTACTGAAGTAGCTAATTTGTTTAGAGGTCA AGAAGATTTGTTGTCAGAATTCGGTCAATTCTTGCCAGAAGCAAAAAGAGGTTCCGGTGCTACTA ACTTCTCTTTGTTGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGGGTAAAAATTAC AAGTCATTGGATTCCGTTGTCGCAAGTGACTTTATTGCCTTGGGTATAACTTCTGAAGTCGCAGA AACATTGCATGGTAGATTAGCCGAAATTGTATGTAACTACGGTGCTGCAACCCCACAAACTTGGA TCAACATAGCAAACCATATCTTGTCACCAGATTTGCCTTTCTCCTTGCACCAAATGTTGTTTTATG GTTGCTACAAGGATTTCGGTCCTGCTCCACCTGCATGGATTCCAGACCCTGAAAAGGTTAAGTC AACTAATTTGGGTGCTTTGTTAGAAAAGAGAGGTAAAGAATTCTTGGGTGTTAAGTACAAGGATC CAATCTCTTCTTTTTCTCACTTCCAAGAATTTTCTGTCAGAAACCCTGAAGTATACTGGAGAACAG TTTTGATGGATGAAATGAAAATAAGTTTCTCTAAGGACCCAGAATGTATCTTGAGAAGAGATGAC ATCAACAACCCAGGTGGTTCTGAATGGTTGCCAGGTGGTTATTTGAACTCAGCTAAAAATTGCTT GAACGTTAACTCCAATAAGAAATTGAATGATACTATGATTGTCTGGAGAGATGAAGGCAACGATG ACTTGCCATTGAATAAGTTGACATTGGATCAATTGAGAAAGAGAGTTTGGTTGGTCGGTTACGCA TTAGAAGAAATGGGTTTGGAAAAAGGTTGTGCCATAGCTATCGATATGCCTATGCATGTAGACGC TGTAGTTATCTATTTGGCTATTGTTTTAGCAGGTTACGTCGTAGTTTCTATAGCTGATTCATTTTCC GCACCAGAAATCTCAACTAGATTGAGATTATCCAAAGCAAAGGCCATATTCACACAAGATCACAT CATCAGAGGTAAAAAGAGAATCCCTTTATACTCAAGAGTCGTAGAAGCCAAATCCCCAATGGCTA TAGTTATCCCTTGTAGTGGTTCTAACATTGGTGCAGAATTAAGAGATGGTGACATATCTTGGGATT ACTTTTTGGAAAGAGCCAAAGAATTCAAGAATTGCGAATTCACTGCCAGAGAACAACCAGTTGAT GCTTACACTAACATTTTGTTCTCCAGTGGTACTACAGGTGAACCAAAAGCAATACCTTGGACACA AGCCACCCCTTTAAAGGCCGCTGCAGATGGTTGGTCACATTTGGATATTAGAAAAGGTGACGTC ATAGTATGGCCAACTAATTTGGGTTGGATGATGGGTCCTTGGTTGGTTTATGCTAGTTTGTTAAAT GGTGCCTCTATTGCTTTATACAACGGTAGTCCATTGGTTTCTGGTTTCGCTAAATTTGTCCAAGAT GCAAAAGTAACAATGTTGGGTGTTGTCCCTTCAATCGTTAGAAGTTGGAAGTCTACAAATTGTGT CTCAGGTTATGATTGGTCCACCATCAGATGCTTTTCTTCATCCGGTGAAGCCTCTAATGTCGACG AATATTTGTGGTTAATGGGTAGAGCTAACTACAAGCCAGTTATCGAAATGTGTGGTGGTACCGAA ATTGGTGGTGCATTCTCAGCCGGTTCCTTTTTACAAGCTCAATCATTGAGTTCTTTTTCATCCCAA TGTATGGGTTGCACATTGTACATCTTGGATAAGAACGGTTACCCAATGCCTAAAAATAAGCCAGG TATTGGTGAATTGGCTTTAGGTCCTGTTATGTTCGGTGCATCTAAAACATTGTTGAACGGTAACCA TCACGATGTATACTTCAAGGGTATGCCAACCTTAAATGGTGAAGTTTTGAGAAGACATGGTGACA TATTCGAATTAACCTCAAACGGTTACTACCATGCCCACGGTAGAGCTGATGACACTATGAACATC GGTGGTATCAAAATCAGTTCTATCGAAATCGAAAGAGTATGTAACGAAGTTGATGACAGAGTCTT TGAAACCACTGCAATTGGTGTTCCACCATTGGGTGGTGGTCCAGAACAATTAGTAATCTTTTTCG TTTTGAAGGATTCTAACGACACAACCATAGATTTGAACCAATTGAGATTATCTTTTAACTTGGGTT TACAAAAGAAATTGAACCCATTATTCAAAGTTACTAGAGTAGTTCCATTGTCATCCTTACCTAGAA CTGCTACAAACAAGATTATGAGAAGAGTCTTGAGACAACAATTCAGTCATTTTGAAGGTTCTGGT

Figure 15B

Complete GPP Gene Cassette Nucleotide Sequence

ATGAAGTTATCTACTAAATTGTGTTGGTGCGGTATTAAGGGTAGATTAAGACCACAAAAGCAACA ACAATTGCATAACACAAACTTGCAAATGACCGAATTGAAGAAACAAAAGACTGCTGAACAAAAGA CTAGACCACAAAACGTTGGTATTAAAGGTATCCAAATCTATATCCCTACACAATGTGTCAATCAAT CTGAATTGGAAAAGTTTGATGGTGTATCACAGGGTAAATACACTATCGGTTTAGGTCAAACAAAC ATGTCTTTCGTAAACGATAGAGAAGACATCTATTCTATGTCATTGACTGTTTTGTCCAAGTTGATA AAAAGTTACAACATCGATACAAACAAGATTGGTAGATTGGAAGTTGGTACCGAAACTTTGATCGA TAAGTCCAAGAGTGTCAAGTCTGTATTGATGCAATTGTTCGGTGAAAATACCGATGTTGAAGGTA TCGACACTTTAAATGCTTGTTATGGTGGTACTAACGCATTATTCAATTCATTGAACTGGATCGAAT CCAATGCCTGGGATGGTAGAGATGCTATTGTTGTCTGCGGTGACATCGCTATCTATGACAAAGG TGCTGCAAGACCAACCGGTGGTGCAGGTACTGTTGCCATGTGGATAGGTCCAGATGCACCTATC GTTTTTGACTCTGTCAGAGCATCATACATGGAACATGCCTACGATTTCTACAAACCAGACTTCAC CTCCGAATATCCTTACGTTGATGGTCACTTTTCTTTGACTTGTTACGTCAAGGCTTTGGACCAAGT ATACAAGTCTTACTCTAAGAAAGCAATATCTAAGGGTTTGGTTTCAGATCCAGCTGGTTCCGACG CATTAAACGTCTTGAAGTACTTCGATTACAACGTTTTCCATGTCCCTACATGCAAGTTGGTTACCA AGTCTTACGGTAGATTGTTGTACAACGATTTCAGAGCTAACCCACAATTGTTCCCTGAAGTCGAC GCTGAATTAGCAACTAGAGATTACGACGAATCTTTGACAGATAAGAACATCGAAAAGACTTTCGT AAACGTTGCAAAGCCATTCCACAAAGAAAGAGTTGCCCAATCATTAATTGTCCCTACAAATACCG GTAACATGTATACAGCCTCAGTTTACGCCGCTTTTGCTTCCTTGTTAAATTATGTAGGTAGTGATG ACTTGCAAGGTAAAAGAGTTGGTTTATTCTCCTATGGTAGTGGTTTAGCAGCCTCTTTGTACTCTT GTAAGATTGTAGGTGACGTTCAACACATTATTAAGGAATTGGACATCACTAATAAGTTGGCTAAG AGAATCACTGAAACACCAAAGGATTATGAAGCTGCAATCGAATTGAGAGAAAACGCACATTTGAA GAAAAATTTCAAACCTCAAGGTAGTATAGAACACTTGCAATCTGGTGTCTACTACTTAACAAACAT CGATGACAAATTCAGAAGATCATACGATGTTAAAAAGAAATTGTCTGGTGGTGGTGGTTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTATAATTTGGGTAACGCTTATTACAA GCAGGGTGACTACCAAAAAGCAATCGAATATTACCAAAAGGCCTTGGAATTAGACCCAAATAACG CCGAAGCTTGGTACAATTTGGGTAATGCATACTATAAACAGGGTGACTATCAAAAGGCTATCGAA TACTACCAAAAGGCATTGGAATTAGACCCTAATAACGCAGAAGCCTGGTACAATTTGGGTAACGC CTACTATAAGCAGGGTGACTATCAAAAAGCCATAGAAGACTACCAAAAGGCTTTGGAATTAGATC CAAATAACTTGCAAGCTGAAGCATGGAAAAATTTGGGTAATGCCTACTACAAACAGGGTGACTAC CAAAAGGCAATTGAATATTATCAAAAAGCCTTGGAATTAGATCCTAATAACGCCTCAGCTTGGTAT AATTTGGGTAATGCCTATTATAAGCAGGGTGACTACCAGAAAGCCATTGAATATTATCAAAAGGC TTTAGAATTGGATCCAAATAACGCAAAAGCCTGGTATAGACGTGGTAATGCCTACTACAAGCAGG GTGACTATCAGAAGGCTATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCAAACAACAGATCC AGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTTGGGT CCTTTGCCACCTGGTTGGGAAGTAAGATCCACAGTTAGTGGTAGAATCTATTTCGTTGATCATAA CAACAGAACTACACAATTCACCGACCCAAGATTGCACGGTTCTGCTGGTTCAGCCGCTGGTTCT GGTGAATTTGGTTCCGCAGAAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCCGGTTCCGCTGGT AGTGCAGCCGGTAGTGGTGAATTTGGTTCTGGTGCTATGGGTCCATTACCACCTGGTTGGGAAA AGAGAACAGATTCTAACGGTAGAGTCTACTTCGTAAACCATAATACCAGAATTACTCAATGGGAA GATCCTAGATCTGGTTCAGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCAGGTGACGTTGAAGA AAATCCAGGTCCAATGGTAGCCGTTAGAAGAAAGGCTTTGTCTATCTTAGCCGAAGCTCCAGTTT TGGCATCAGATAGATTACCTTACAAGAACTACGATTACGACAGAGTATTTGGTGCTTGTTGCGAA AACGTTATTGGTTATATGCCATTGCCTGTCGGTGTAATCGGTCCATTAGTTATTGATGGTACATCT TACCATATCCCTATGGCAACTACAGAAGGTTGTTTGGTTGCATCAGCCATGAGAGGTTGCAAGG CAATTAATGCTGGTGGTGGTGCTACCACTGTTTTAACCAAAGATGGTATGACTAGAGGTCCAGTT GTCAGATTTCCTACTTTGAAGAGATCCGGTGCTTGTAAAATATGGTTAGATAGTGAAGAAGGTCA AAATGCCATCAAAAAGGCTTTTAACTCCACAAGTAGATTCGCAAGATTGCAACATATTCAAACATG CTTAGCCGGTGACTTGTTGTTTATGAGATTCAGAACAACCACTGGTGACGCTATGGGTATGAATA TGATATCTAAGGGTGTCGAATACTCATTGAAGCAAATGGTAGAAGAATACGGTTGGGAAGATATG GAAGTAGTTTCTGTTTCAGGCAACTACTGTACTGACAAAAAGCCAGCTGCAATTAACTGGATAGA

AGGTCGTGGTAAATCTGTCGTAGCTGAAGCAACAATACCTGGTGACGTTGTTAGAAAGGTTTTGA AATCTGACGTATCAGCTTTGGTTGAATTGAACATCGCTAAAAATTTGGTTGGTTCCGCCATGGCT GGTAGTGTCGGTGGTTTTAATGCACATGCCGCTAACTTAGTTACAGCAGTCTTCTTGGCCTTAGG TCAAGATCCAGCTCAAAACGTAGAATCTTCAAACTGTATCACCTTGATGAAAGAAGTTGATGGTG ACTTAAGAATATCCGTTAGTATGCCATCAATAGAAGTCGGTACAATCGGTGGTGGTACCGTCTTG GAACCTCAAGGTGCAATGTTAGATTTGTTAGGTGTTAGAGGTCCACATGCAACTGCCCCTGGTA CAAATGCTAGACAATTGGCAAGAATTGTCGCTTGTGCAGTATTAGCTGGTGAATTGTCCTTATGC GCAGCCTTGGCTGCAGGTCACTTAGTTCAAAGTCATATGACACACAACAGAAAGTTGTCTGGTG GTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCTTGGTATAATTTGGG TAACGCATATTACAAGCAGGGTGACTACCAAAAGGCCATCGAATACTACCAAAAGGCTTTGGAAT TGGACCCAAATAACGCAGAAGCCTGGTACAATTTGGGTAATGCTTACTATAAACAGGGTGACTAT CAAAAGGCAATTGAATATTACCAAAAGGCCTTGGAATTAGACCCTAATAACGCTGAAGCATGGTA CAATTTGGGTAACGCCTACTATAAGCAGGGTGACTATCAAAAAGCTATTGAAGACTACCAAAAGG CATTGGAATTAGATCCAAATAACTTGCAAGCCGAAGCTTGGAAAAATTTGGGTAACGCTTACTAC AAACAGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAAGCTTTGGAATTGGACCCTAATAA CGCATCTGCCTGGTATAATTTGGGTAATGCTTATTATAAACAGGGTGACTACCAGAAGGCAATAG AATACTATCAAAAAGCCTTGGAATTAGACCCAAATAACGCTAAAGCATGGTATAGACGTGGTAAT GCTTACTATAAGCAGGGTGACTACCAGAAAGCTATAGAAGATTATCAAAAGGCATTGGAATTGGA TCCTAACAACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGT GGTGCTTCCAGTTATTACCATCACCATCACCATCACTTGGAATCCACAAGTTTATACAAAAAGGC AGGTTCAGAATTTTTCAGAAGAGAAAGAAATAAGATGGCCGCTGCAAAATGTAGAAACAGAAGAA GAGAATTGACAGATACCTTACAAGCTGAAACCGATCAATTGGAAGACGAAAAGTCTGCATTGCAA ACTGAAATAGCCAATTTGTTGAAGGAAAAGGAAAAGTTGGAATTCATTTTAGCCGCTCATAGACC AGCTTGCAAAATTCCTGATGACTTGGGTTTCCCAGAAGAAATGTCTTTAGAAGGTTCCGCAGGTA GTGCAGCCGGTTCCGGTGAATTTGGTAGTGCTGAAGCTGCAGCCAAGGAAGCTGCAGCCAAAG CTGGTTCTGCAGGTTCAGCTGCAGGTTCCGGTGAATTCGGTTCTTCATACTATCACCATCACCAT CATCACTTGGAATCTACCTCATTATACAAAAAGGCTGGTTCCGGTAGTCAAAAGGTTGAATCTTT GAAGCAAAAGATTGAAGAATTGAAGCAAAGAAAAGCCCAATTGAAGAATGATATCGCTAACTTAG AAAAGGAAATCGCCTACGCTGAAACTGGTTCTGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCA GGTGACGTTGAAGAAAATCCAGGTCCAATGAGTTTACCATTTTTGACATCTGCTCCTGGTAAAGT TATTATATTCGGTGAACATAGTGCCGTCTATAATAAGCCAGCTGTCGCTGCATCTGTATCAGCTTT GAGAACATACTTGTTGATCTCTGAATCTTCAGCACCTGATACCATCGAATTGGATTTCCCAGACA TCTCATTCAATCACAAGTGGTCCATTAATGATTTCAACGCTATCACCGAAGACCAAGTAAACTCAC AAAAGTTGGCCAAAGCTCAACAAGCAACTGATGGTTTGTCACAAGAATTAGTTTCCTTGTTAGAC CCATTGTTGGCTCAATTGTCCGAAAGTTTCCATTACCACGCCGCTTTCTGTTTCTTGTACATGTTC GTTTGTTTATGCCCTCATGCTAAGAATATCAAATTTTCTTTGAAGTCTACTTTGCCAATTGGTGCA GGTTTAGGTTCCAGTGCCTCTATATCAGTTTCCTTAGCATTGGCCATGGCTTATTTGGGTGGTTT GATAGGTAGTAACGATTTGGAAAAGTTGTCTGAAAACGACAAGCATATCGTCAACCAATGGGCAT TCATCGGTGAAAAATGCATTCACGGTACTCCTAGTGGTATAGATAATGCAGTTGCCACATATGGT AACGCTTTGTTATTCGAAAAGGACTCTCATAACGGTACCATCAACACTAACAACTTCAAGTTCTTG GATGACTTTCCTGCAATACCAATGATCTTGACTTACACAAGAATTCCAAGATCTACTAAAGATTTG GTAGCTAGAGTCAGAGTATTGGTTACAGAAAAGTTCCCTGAAGTTATGAAGCCAATCTTGGATGC AATGGGTGAATGTGCCTTGCAAGGTTTGGAAATCATGACAAAGTTGTCAAAGTGCAAGGGTACT GATGACGAAGCTGTTGAAACAAATAACGAATTGTACGAACAATTGTTGGAATTGATCAGAATCAA TCATGGTTTGTTAGTTTCAATTGGTGTCTCCCACCCAGGTTTAGAATTGATAAAGAACTTGTCAGA TGACTTAAGAATCGGTTCCACAAAATTGACCGGTGCTGGTGGTGGTGGTTGTTCTTTGACCTTGT TAAGAAGAGATATCACTCAAGAACAAATCGACAGTTTTAAAAAGAAATTGCAAGATGACTTCTCTT ACGAAACTTTCGAAACAGATTTGGGTGGTACTGGTTGTTGCTTGTTGTCAGCTAAGAATTTGAAC AAAGATTTGAAGATTAAATCCTTGGTTTTCCAATTGTTCGAAAATAAGACTACAACCAAGCAACAA ATCGATGACTTGTTGTTGCCTGGTAATACAAACTTGCCATGGACCTCAAAATTATCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTATAATTTGGGTAAC GCATATTACAAGCAGGGTGACTACCAAAAGGCTATCGAATACTACCAAAAGGCATTGGAATTGGA

CCCTAATAACGCCGAAGCTTGGTACAATTTGGGTAATGCTTACTATAAACAGGGTGACTATCAAA AGGCCATTGAATATTACCAAAAGGCTTTGGAATTGGACCCAAATAACGCAGAAGCCTGGTACAAT TTGGGTAACGCTTACTATAAGCAGGGTGACTATCAAAAAGCAATTGAAGACTACCAAAAGGCCTT AGAATTGGATCCTAATAACTTGCAAGCTGAAGCATGGAAAAATTTGGGTAACGCTTATTATAAACA GGGTGACTACCAAAAAGCCATTGAATACTATCAAAAAGCATTGGAATTGGATCCAAATAACGCCT CTGCTTGGTATAATTTGGGTAATGCTTATTATAAGCAGGGTGACTACCAGAAAGCCATAGAATAC TATCAAAAAGCTTTGGAATTAGACCCTAATAACGCAAAAGCCTGGTATAGACGTGGTAATGCTTA CTACAAACAGGGTGACTATCAGAAGGCAATAGAAGATTATCAAAAAGCTTTAGAATTAGACCCAA ATAACAGAAGTAGATCTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTG CTTCTATGGAACCTGCAATGGAACCAGAAACATTGGAAGCCAGAATCAATAGAGCTACCAATCCT TTGAACAAGGAATTGGATTGGGCTTCTATTAATGGTTTCTGTGAACAATTGAACGAAGACTTCGA AGGTCCACCTTTAGCAACAAGATTATTGGCCCATAAAATTCAATCACCACAAGAATGGGAAGCAA TACAAGCCTTAACCGTCTTGGAAACTTGTATGAAGTCCTGCGGTAAAAGATTCCACGATGAAGTT GGTAAATTCAGATTTTTGAACGAATTGATCAAGGTTGTCTCACCTAAGTATTTGGGTAGTAGAACA TCTGAAAAGGTTAAAAACAAGATCTTGGAATTGTTGTACTCCTGGACCGTAGGTTTACCAGAAGA AGTTAAGATCGCTGAAGCATACCAAATGTTGAAGAAACAAGGTATTGTTAAGTCAGGTTCCGCCG GTAGTGCAGCCGGTTCTGGTGAATTCGGTTCTGCAGAAGCTGCAGCCAAGGAAGCTGCAGCCA AAGCTGGTTCAGCAGGTTCCGCTGCAGGTTCTGGTGAATTTGGTTCAGGTGCAATGGGTTCCAT GGCCGAAGCTGAAGGTGAAAGTTTGGAATCTTGGTTAAATAAGGCTACAAATCCATCAAACAGAC AAGAAGATTGGGAATATATCATTGGTTTCTGTGACCAAATCAATAAGGAATTGGAAGGTCCTCAA ATAGCTGTTAGATTATTGGCACATAAGATCCAATCTCCACAAGAATGGGAAGCCTTACAAGCTTT GACTGTTTTAGAAGCTTGTATGAAGAATTGCGGTAGAAGATTTCACAACGAAGTCGGTAAATTCA GATTTTTGAATGAATTAATTAAGGTAGTTAGTCCAAAATACTTAGGTGACAGAGTTTCTGAAAAGG TTAAGACCAAAGTCATAGAATTGTTGTACTCTTGGACTATGGCCTTGCCTGAAGAAGCTAAGATC AAAGATGCATACCATATGTTGAAGAGACAAGGTATAGTCCAATCAGATCCACCTATCCCAGTAGA CAGAACTTTGATTCCATCTCCACCACCAAGACCTAAAAATGGTTCCGGTGCTACTAACTTCTCTTT GTTGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGTCCGAATTAAGAGCTTTTAGT GCACCTGGTAAAGCCTTGTTAGCTGGTGGTTATTTGGTTTTGGATACAAAGTACGAAGCATTCGT TGTCGGTTTGTCAGCCAGAATGCATGCAGTCGCCCACCCTTACGGTTCTTTACAAGGTTCTGATA AGTTCGAAGTAAGAGTCAAGTCTAAGCAATTCAAGGACGGTGAATGGTTATACCATATATCTCCA AAGTCAGGTTTTATTCCTGTTTCCATAGGTGGTAGTAAAAATCCATTCATCGAAAAGGTTATTGCA AACGTCTTTTCTTACTTCAAGCCTAACATGGATGACTACTGTAACAGAAACTTGTTCGTCATCGAT ATATTCTCTGATGACGCTTATCATTCTCAAGAAGACTCAGTAACTGAACACAGAGGTAATAGAAG ATTGTCCTTTCATAGTCACAGAATTGAAGAAGTTCCAAAAACCGGTTTAGGTTCTTCAGCTGGTG GTTTAGTCACTGTATTGACTACAGCTTTAGCATCCTTTTTCGTTAGTGATTTGGAAAACAACGTAG ACAAGTACAGAGAAGTTATTCATAATTTGGCACAAGTAGCCCACTGCCAAGCACAAGGTAAAATC GGTTCCGGTTTTGATGTTGCTGCAGCCGCTTATGGTTCAATTAGATACAGAAGATTCCCACCTGC TTTGATATCTAATTTGCCAGATATCGGTTCTGCTACATATGGTTCAAAGTTGGCACATTTGGTTGA TGAAGAAGACTGGAACATCACAATTAAATCCAACCATTTGCCTAGTGGTTTGACCTTATGGATGG GTGACATTAAGAATGGTTCTGAAACTGTTAAGTTGGTCCAAAAAGTAAAGAACTGGTACGATTCT CATATGCCAGAATCATTGAAGATCTACACAGAATTAGACCATGCTAATTCCAGATTCATGGATGG TTTGAGTAAATTAGACAGATTGCATACCCACGATGACTACTCTGATCAAATCTTCGAATCATTGGA AAGAAACGACTGTACTTGCCAAAAATACCCAGAAATCACAGAAGTAAGAGATGCCGTTGCTACCA TAAGAAGATCTTTTAGAAAGATCACTAAGGAATCAGGTGCAGATATCGAACCACCTGTTCAAACA TCTTTGTTAGATGACTGTCAAACCTTGAAGGGTGTCTTAACTTGCTTGATTCCAGGTGCTGGTGG TTATGATGCAATAGCCGTCATCACTAAACAAGATGTAGACTTGAGAGCTCAAACAGCAAACGATA AGAGATTTTCAAAGGTCCAATGGTTAGATGTAACCCAAGCTGACTGGGGTGTTAGAAAAGAAAAG GATCCTGAAACTTACTTGGACAAAAAGTTATCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTG GTGGTGGTGGTAGTGCTGAAGCATGGTACAATTTGGGTAACGCATACTACAAGCAGGGTGACTA CCAAAAGGCCATAGAATACTACCAAAAGGCTTTGGAATTGGACCCAAATAACGCCGAAGCTTGG TATAATTTGGGTAATGCTTATTATAAACAGGGTGACTATCAAAAGGCAATCGAATACTACCAAAAG GCCTTGGAATTAGACCCTAATAACGCAGAAGCCTGGTATAATTTGGGTAACGCTTATTATAAGCA

GGGTGACTATCAAAAAGCTATCGAAGACTACCAAAAGGCATTGGAATTAGATCCAAATAACTTGC AAGCTGAAGCATGGAAGAATTTGGGTAACGCTTACTATAAACAGGGTGACTACCAAAAAGCCATT GAATATTATCAAAAAGCTTTGGAATTGGATCCTAATAACGCCTCTGCTTGGTACAATTTGGGTAAT GCTTACTATAAGCAGGGTGACTATCAGAAGGCTATTGAATATTATCAAAAGGCTTTAGAATTGGA CCCTAATAACGCAAAGGCCTGGTACAGACGTGGTAACGCTTATTACAAACAGGGTGACTACCAG AAAGCTATTGAAGATTATCAAAAGGCATTGGAATTGGATCCTAACAACAGATCCAGAAGTGCTGG TGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCCAGTTATTACCATCACCAT CACCATCACTTGGAATCTACATCATTATACAAAAAGGCTGGTTCCGGTAGTCAAAAGGTTGAAGA ATTGAAAAATAAGATAGCCGAATTGGAAAACAGAAACGCTGTTAAAAAGAACAGAGTCGCACATT TGAAACAAGAAATAGCCTACTTGAAGGATGAATTAGCAGCCCATGAATTTGAAGGTTCTGCCGGT TCAGCTGCAGGTTCTGGTGAATTCGGTTCAGCTGAAGCCGCTGCAAAAGAAGCCGCTGCAAAG GCCGGTTCCGCTGGTAGTGCCGCTGGTTCTGGTGAATTTGGTTCTTCATACTATCACCATCACCA TCATCACTTGGAATCTACTTCATTATATAAAAAGGCCGGTTCCGGTAGTTTCGAAAACGTTACACA TGAATTCATTTTGGCTACCTTGGAAAACGAAAACGCAAAGTTAAGAAGATTGGAAGCCAAGTTGG AAAGAGAATTAGCTAGATTGAGAAATGAAGTTGCATGGTTAGGTTCTGGTGCTACTAACTTCTCT TTGTTGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGACAGTTTATACCGCTTCTGT CACCGCACCTGTAAATATTGCTACTTTGAAATACTGGGGTAAAAGAGATACTAAGTTGAATTTGC CAACAAACTCTTCAATCTCAGTTACATTGTCCCAAGATGACTTAAGAACCTTGACTTCTGCTGCAA CTGCTCCTGAATTCGAAAGAGATACATTGTGGTTGAATGGTGAACCACATTCTATCGACAACGAA AGAACTCAAAACTGTTTGAGAGATTTGAGACAATTGAGAAAGGAAATGGAGAGTAAGGATGCTTC TTTGCCTACATTGAGTCAATGGAAGTTGCACATAGTTTCTGAAAACAACTTCCCAACCGCCGCTG GTTTGGCATCCAGTGCAGCCGGTTTCGCTGCATTAGTCTCTGCAATCGCCAAGTTGTACCAATTG CCACAAAGTACATCTGAAATCAGTAGAATCGCTAGAAAAGGTTCAGGTTCCGCATGTAGATCTTT ATTTGGTGGTTACGTCGCATGGGAAATGGGTAAAGCCGAAGACGGTCATGATTCAATGGCCGTA CAAATAGCTGACTCTTCAGATTGGCCTCAAATGAAAGCTTGCGTCTTGGTTGTCTCAGACATCAA AAAGGATGTATCCAGTACACAAGGCATGCAATTGACTGTTGCAACATCCGAATTGTTTAAAGAAA GAATCGAACACGTAGTTCCAAAAAGATTCGAAGTCATGAGAAAGGCTATCGTAGAAAAGGATTTC GCCACCTTCGCTAAGGAAACTATGATGGACAGTAACTCTTTCCATGCAACTTGTTTGGATTCATTT CCACCTATTTTCTATATGAACGACACCTCAAAGAGAATAATCTCCTGGTGCCACACTATCAACCA ATTCTACGGTGAAACAATCGTTGCTTACACCTTCGATGCAGGTCCTAATGCCGTCTTGTATTACTT AGCCGAAAACGAATCAAAGTTGTTCGCTTTTATATATAAGTTGTTTGGTTCCGTTCCAGGTTGGG ATAAAAAGTTCACTACAGAACAATTGGAAGCTTTTAATCATCAATTCGAATCTTCAAACTTTACTG CCAGAGAATTGGACTTAGAATTGCAAAAGGATGTAGCTAGAGTTATCTTGACCCAAGTTGGTTCA GGTCCTCAAGAAACTAACGAATCCTTGATAGATGCTAAGACAGGTTTGCCAAAAGAAAAATTGTC TGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTATAAT TTGGGTAACGCTTATTACAAGCAGGGTGACTACCAAAAGGCCATCGAATACTACCAAAAGGCTTT GGAATTGGACCCTAATAACGCCGAAGCTTGGTACAATTTGGGTAATGCCTACTATAAACAGGGT GACTATCAAAAAGCAATTGAATATTACCAAAAGGCCTTGGAATTGGACCCAAATAACGCAGAAGC CTGGTACAATTTGGGTAACGCCTACTATAAGCAGGGTGACTATCAAAAGGCTATCGAAGATTACC AAAAGGCATTAGAATTGGATCCTAATAACTTGCAAGCTGAAGCATGGAAAAATTTGGGTAATGCC TATTATAAACAGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAAGCTTTAGAATTAGACCCA AATAACGCCTCAGCTTGGTATAATTTGGGTAATGCATACTACAAACAGGGTGACTATCAGAAGGC AATTGAATACTATCAAAAGGCATTAGAATTAGATCCTAATAACGCAAAAGCCTGGTATAGACGTG GTAATGCCTACTACAAGCAGGGTGACTATCAGAAGGCGATTGAAGACTACCAAAAGGCATTGGA ATTGGATCCAAACAACAGATCAAGATCCGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGT GGTGGTGGTGCTTCTGCAATGGCCGATTTGGAACAAAAGGTATTGGAAATGGAAGCTAGTACAT ATGACGGTGTTTTTATTTGGAAGATCTCTGATTTCCCAAGAAAAAGACAAGAAGCTGTTGCAGGT AGAATCCCTGCTATTTTTAGTCCAGCATTCTACACCTCTAGATACGGTTACAAGATGTGTTTGAGA ATATATTTGAATGGTGACGGTACTGGTAGAGGTACTCATTTGTCTTTGTTTTTCGTCGTAATGAAG GGTCCTAATGATGCTTTGTTGAGATGGCCTTTTAATCAAAAGGTTACCTTGATGTTGTTGGATCAA AACAACAGAGAACACGTTATCGACGCTTTTAGACCTGATGTCACTTCCAGTTCTTTCCAAAGACC AGTTAATGATATGAACATTGCTTCTGGTTGTCCTTTGTTTTGCCCAGTCTCAAAGATGGAAGCTAA

AAATTCCTATGTTAGAGATGACGCCATCTTCATTAAGGCTATCGTTGATTTGACTGGTTTAGGTTC AGCAGGTTCCGCCGCTGGTTCTGGTGAATTTGGTTCCGCCGAAGCAGCCGCTAAGGAAGCAGC CGCTAAAGCAGGTAGTGCCGGTTCTGCAGCCGGCTCTGGCGAATTTGGTAGTGCCTCTATTAAA TTGCAATCATCCGACGGTGAAATCTTCGAAGTTGATGTCGAAATAGCAAAGCAATCTGTTACCAT AAAAACTATGTTGGAAGATTTGGGTATGGATGACGAAGGTGACGATGATCCAGTTCCTTTGCCAA ATGTCAACGCTGCAATATTGAAGAAAGTTATTCAATGGTGCACACATCACAAGGACGATCCACCT CCACCTGAAGACGATGAAAATAAGGAAAAGAGAACTGACGATATTCCAGTATGGGACCAAGAAT TCTTGAAGGTTGATCAAGGTACATTGTTCGAATTGATCTTGGCCGCTAACTATTTGGACATCAAG GGTTTGTTAGATGTAACATGTAAAACCGTTGCTAACATGATCAAGGGTAAAACACCAGAAGAAAT CAGAAAGACCTTTAATATTAAGAATGATTTCACTGAAGAAGAAGAAGCACAAGTTAGAAAGGAAA ACCAATGGTGCGGTTCTGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCAGGTGACGTTGAAGAA AATCCAGGTCCAATGACTGCTGATAATAACTCTATGCCACATGGTGCCGTATCTTCATACGCTAA GTTGGTTCAAAACCAAACACCTGAAGATATCTTGGAAGAATTCCCAGAAATCATCCCTTTGCAAC AAAGACCAAACACTAGATCCAGTGAAACATCCAACGATGAAAGTGGTGAAACCTGTTTTTCAGGT CATGACGAAGAACAAATTAAATTGATGAACGAAAACTGCATCGTATTGGATTGGGATGACAATGC AATAGGTGCCGGTACTAAGAAAGTTTGTCATTTGATGGAAAACATAGAAAAGGGTTTGTTGCACA GAGCTTTCTCCGTTTTTATATTCAATGAACAGGGTGAATTGTTATTGCAACAAAGAGCAACAGAAA AGATCACCTTTCCAGATTTGTGGACTAATACATGTTGCTCTCATCCTTTGTGCATTGATGACGAAT TAGGTTTGAAGGGTAAATTGGATGACAAAATTAAGGGTGCTATAACTGCTGCAGTCAGAAAATTA GATCATGAATTGGGTATACCAGAAGACGAAACCAAGACTCGTGGTAAATTCCATTTCTTAAACAG AATCCACTATATGGCTCCATCTAACGAACCTTGGGGTGAACATGAAATCGATTACATCTTATTTTA CAAGATTAATGCAAAGGAAAACTTGACAGTTAACCCAAACGTTAATGAAGTCAGAGATTTCAAAT GGGTTTCTCCTAATGATTTGAAGACCATGTTTGCTGACCCATCATATAAGTTTACTCCTTGGTTCA AGATCATCTGTGAAAACTACTTGTTTAACTGGTGGGAACAATTAGATGACTTGTCTGAAGTTGAAA ACGATAGACAAATCCATAGAATGTTGAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCT GGTGGTGGTGGTAGTGCCGAAGCTTGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACT ACCAAAAGGCAATCGAATACTACCAAAAGGCCTTGGAATTGGACCCAAATAACGCAGAAGCCTG GTATAATTTGGGTAATGCATATTATAAACAGGGTGACTATCAAAAGGCTATTGAATATTACCAAAA GGCATTGGAATTGGACCCTAATAACGCTGAAGCATGGTATAATTTGGGTAACGCCTATTATAAGC AGGGTGACTATCAAAAAGCCATCGAAGACTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTA CAAGCCGAAGCTTGGAAGAATTTGGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTAT TGAATACTATCAAAAAGCCTTAGAATTAGACCCTAATAACGCATCTGCCTGGTACAATTTGGGTAA TGCCTATTACAAGCAGGGTGACTATCAGAAGGCTATTGAATACTACCAAAAAGCATTGGAATTGG ATCCAAATAACGCTAAGGCATGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTATCAA AAGGCGATTGAAGATTATCAAAAAGCTTTGGAATTGGATCCTAACAACAGATCTAGATCAGCTGG TGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCATATTACCATCACCAT CACCATCACTTAGAATCCACAAGTTTGTACAAAAAGGCTGGTTCTGGTTCAAACACCGTTAAGGA ATTAAAGAACTACATCCAAGAATTGGAAGAAAGAAACGCAGAATTGAAAAATTTGAAGGAACATTT GAAGTTTGCCAAGGCTGAATTAGAATTCGAATTGGCCGCTCACAAATTTGAAGGTTCCGCTGGTA GTGCAGCCGGTTCCGGTGAATTCGGTAGTGCAGAAGCTGCAGCCAAAGAAGCTGCAGCCAAGG CTGGTTCTGCAGGTTCAGCTGCAGGTTCTGGTGAATTTGGTTCCAGTTACTATCACCATCACCAT CATCACTTAGAATCCACAAGTTTGTATAAAAAGGCCGGTTCTGGTTCACAAAAAGTCGCACAATT AAAGAATAGAGTAGCCTACAAGTTGAAGGAAAACGCTAAGTTGGAAAACATTGTCGCAAGATTAG AAAACGATAATGCCAACTTGGAAAAAGACATCGCTAATTTGGAAAAGGATATTGCAAACTTGGAA AGAGATGTTGCCAGAGGTTCTGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCAGGTGACGTTGA AGAAAATCCAGGTCCAATGGAAGCTAAGATAGATGAATTGATAAATAACGACCCAGTTTGGTCTT CACAAAACGAATCCTTGATCAGTAAGCCATACAACCATATCTTGTTAAAACCTGGTAAAAATTTCA GATTAAATTTGATCGTACAAATCAACAGAGTTATGAATTTGCCTAAGGATCAATTGGCTATCGTTT CTCAAATAGTCGAATTGTTGCATAACTCCAGTTTGTTGATCGATGACATCGAAGATAACGCACCA TTGAGAAGAGGTCAAACTACATCCCACTTAATTTGGGGTGTCCCTAGTACTATTAATACCGCAAA CTACATGTACTTCAGAGCCATGCAATTGGTATCACAATTGACCACTAAGGAACCATTGTACCATT GGTTGATCACAATTTTTAACGAAGAATTGATTAATTTGCACAGAGGTCAAGGTTTGGATATCTATT

GGAGAGACTTCTTACCAGAAATTATACCTACCCAAGAAATGTACTTGAACATGGTAATGAATAAG ACTGGTGGTTTGTTTAGATTGACCTTGAGATTAATGGAAGCTTTGTCTCCATCTTCACATCACGGT CATTCATTGGTTCCTTTCATAAACTTGTTGGGTATCATCTATCAAATCAGAGATGACTACTTGAATT TGAAGGATTTCCAAATGTCCAGTGAAAAGGGTTTCGCAGAAGACATAACTGAGGGTAAATTGTCA TTCCCAATCGTCCATGCCTTAAACTTCACAAAAACCAAGGGTCAAACCGAACAACACAATGAAAT CTTAAGAATTTTGTTATTGAGAACTTCTGATAAGGACATAAAGTTGAAGTTGATCCAAATCTTGGA ATTCGATACCAACTCATTGGCTTACACTAAGAACTTCATCAACCAATTGGTTAACATGATTAAGAA TGATAACGAAAATAAGTACTTGCCAGATTTGGCCTCCCATAGTGACACTGCTACAAATTTGCACG ATGAATTGTTGTACATCATCGACCATTTGTCCGAATTGAAATTATCTGGTGGTGGTGGTTCTGGT GGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTACAACTTGGGTAACGCTTACTACA AGCAGGGTGACTACCAAAAGGCTATCGAATACTACCAAAAGGCATTGGAATTAGACCCAAATAAC GCTGAAGCATGGTACAACTTAGGCAACGCATATTATAAACAGGGTGACTATCAAAAGGCCATAGA ATACTACCAAAAGGCTTTGGAATTGGACCCTAATAACGCCGAAGCTTGGTACAACTTGGGTAATG CTTATTACAAGCAGGGTGACTATCAAAAAGCAATTGAAGACTACCAAAAAGCCTTGGAATTAGAT CCAAATAACTTGCAAGCAGAAGCCTGGAAGAACTTAGGCAACGCATACTATAAACAGGGTGACT ACCAAAAAGCCATTGAATATTATCAAAAAGCTTTGGAATTAGACCCTAATAACGCTTCTGCTTGGT ATAACTTAGGCAATGCCTATTATAAGCAGGGTGACTATCAGAAAGCTATTGAATATTATCAAAAGG CCTTGGAATTGGACCCAAATAACGCCAAGGCTTGGTACAGACGTGGTAACGCATACTACAAACA GGGTGACTATCAGAAGGCTATCGAAGATTATCAAAAAGCATTAGAATTAGATCCTAATAACAGAT CTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTTGTG TACTATGAAAAAGGGTCCATCTGGTTACGGTTTTAATTTGCATTCTGATAAGTCAAAGCCTGGTCA ATTCATAAGATCAGTTGATCCAGACTCCCCTGCAGAAGCCAGTGGTTTGAGAGCTCAAGATAGAA TTGTCGAAGTAAATGGTGTCTGCATGGAAGGTAAACAACACGGTGACGTTGTTTCTGCTATTAGA GCTGGTGGTGACGAAACTAAGTTATTGGTAGTTGACAGAGAAGGTTCCGCCGGTAGTGCTGCAG GTTCTGGTGAATTTGGTTCAGCTGAAGCCGCTGCAAAAGAAGCCGCTGCAAAGGCCGGTTCTGC TGGTTCAGCCGCTGGTTCTGGTGAATTCGGTTCTTCATCCGGTGCTATAATCTATACAGTTGAAT TGAAGAGATACGGTGGTCCATTAGGTATTACTATATCTGGTACAGAAGAACCATTCGATCCTATC ATCATCAGTTCTTTGACTAAGGGTGGTTTAGCTGAAAGAACAGGTGCAATCCATATTGGTGACAG AATATTGGCTATCAATTCATCCAGTTTGAAAGGTAAACCATTGTCAGAAGCTATCCACTTATTGCA AATGGCAGGTGAAACCGTTACTTTGAAAATCAAAAAGCAAACAGATGCACAACCTGCCTCTTCAG GTTCTGGT

Figure 15C

Complete CAN Gene Cassette Nucleotide Sequence

ATGAATCATTTGAGAGCCGAAGGACCAGCTTCTGTCTTAGCAATAGGTACTGCCAATCCAGAGAA CATCTTGTTACAAGATGAATTTCCTGACTATTACTTCAGAGTTACCAAATCCGAGCATATGACGCA GTTGAAGGAAAAGTTTAGAAAGATCTGTGATAAGAGTATGATCAGAAAGAGGAACTGCTTCTTAA ACGAAGAGCATTTGAAGCAAAATCCTAGATTAGTGGAACACGAGATGCAAACATTGGATGCTAG GCAGGACATGTTAGTTGTCGAAGTTCCTAAATTGGGTAAAGATGCATGTGCCAAAGCTATTAAGG AATGGGGTCAACCCAAGTCTAAGATAACTCATTTGATTTTTACAAGTGCTAGCACTACAGATATG CCTGGTGCAGACTATCACTGTGCCAAACTACTTGGTTTATCGCCCTCTGTGAAGAGAGTTATGAT GTATCAACTAGGTTGCTACGGTGGTGGTACTGTACTTAGAATCGCTAAAGACATTGCAGAAAATA ACAAGGGTGCCAGGGTCTTGGCTGTATGTTGCGATATTATGGCTTGCTTGTTTAGAGGTCCATCA GAATCCGATTTGGAGCTGTTGGTTGGTCAAGCTATTTTCGGTGACGGTGCTGCAGCTGTTATTGT TGGTGCAGAACCTGATGAGTCAGTCGGTGAAAGACCAATCTTTGAATTGGTTTCTACCGGTCAAA CGATTTTACCAAATAGTGAAGGTACAATAGGTGGTCATATCAGAGAAGCTGGTTTGATATTCGAT TTGCACAAAGACGTTCCTATGCTAATATCTAACAACATCGAAAAGTGTCTGATCGAGGCTTTTAC

CCCCATCGGTATTTCCGATTGGAATAGTATATTCTGGATCACGCATCCAGGTGGTAAAGCAATCC TGGATAAGGTTGAAGAGAAGCTGCATTTGAAGTCTGATAAGTTTGTCGACAGCAGACATGTATTG TCGGAACACGGTAACATGTCTTCATCCACAGTGCTGTTCGTTATGGATGAACTTAGAAAGAGATC TTTGGAAGAGGGTAAAAGCACCACGGGTGACGGTTTTGAATGGGGTGTTCTTTTTGGATTCGGC CCCGGTTTGACCGTCGAAAGAGTAGTTGTTAGATCTGTACCAATTAAATACAAGTTGTCTGGTGG TGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTACAATTTGGGT AACGCTTACTACAAGCAGGGTGACTACCAGAAGGCTATCGAGTATTACCAAAAAGCACTTGAACT GGATCCAAATAACGCTGAGGCATGGTATAATTTGGGCAACGCATATTACAAACAGGGTGACTATC AAAAGGCCATAGAATACTACCAAAAGGCTTTGGAGCTGGATCCTAATAACGCCGAAGCTTGGTA CAATTTGGGAAATGCCTATTATAAGCAGGGTGACTATCAGAAGGCAATAGAGGACTACCAAAAAG CCCTAGAACTTGATCCAAATAATTTGCAGGCAGAAGCCTGGAAGAATTTGGGTAATGCTTACTAT AAACAGGGTGACTATCAGAAAGCTATTGAATACTACCAAAAAGCACTGGAATTGGATCCTAATAA CGCTTCTGCTTGGTACAATTTGGGCAACGCTTACTACAAACAGGGTGACTACCAAAAAGCTATCG AATATTATCAAAAGGCTCTGGAACTAGATCCAAATAACGCCAAGGCTTGGTATAGAAGGGGAAAT GCTTATTATAAACAGGGTGACTACCAGAAAGCAATTGAAGACTACCAAAAAGCCCTTGAACTGGA TCCTAATAACAGATCTAGAAGCGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGT GGTGCTTCTGGTAACAACTTAGAAACATACGAGTGGTACAATAAGTCTATTTCTAGAGATAAGGC CGAAAAGTTACTACTTGACACCGGTAAAGAAGGTGCTTTTATGGTTAGAGATTCTAGAACTCCAG GTACTTATACAGTCTCTGTATTCACAAAGGCTATCATCTCAGAAAACCCATGTATCAAGCATTACC ACATCAAGGAAACCAACGACTCTCCTAAAAGATATTACGTGGCAGAAAAGTACGTTTTTGATTCA ATCCCACTGTTGATTCAATATCATCAGTACAATGGTGGTGGTTTGGTGACTAGATTGAGGTATCC TGTTTGCGGTGGTAGCGCAGGTTCGGCTGCAGGATCAGGCGAATTTGGTTCCGCCGAGGCCGC TGCAAAAGAAGCCGCTGCAAAGGCTGGATCTGCAGGCTCAGCCGCTGGTTCTGGAGAATTTGG TTCTGGTTCTCATCCCTGGTTTTTCGGTAAAATTCCAAGAGCAAAGGCCGAAGAAATGTTGTCTA AACAAAGACACGACGGTGCATTTTTGATAAGGGAAAGTGAGAGCGCACCTGGTGACTTTTCGTT GTCTGTTAAATTCGGTAATGATGTCCAACATTTCAAGGTATTGAGAGATGGTGCTGGTAAATACTT TTTGTGGGTCGTAAAGTTCAATTCCTTGAACGAATTAGTGGATTACCATAGATCAACTTCCGTTAG TAGGAACCAACAGATTTTCTTGAGAGATATCGAACAAGTTCCACAACAGCCTACAGGTTCTGGAG CTACTAACTTCTCTTTGTTGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGGCTGTA AAGCATTTGATCGTGTTGAAATTCAAGGATGAAATCACAGAGGCACAAAAGGAAGAGTTTTTCAA GACCTACGTTAATTTGGTCAACATAATCCCAGCTATGAAAGATGTATACTGGGGTAAAGACGTGA CCCAAAAGAATAAGGAAGAGGGTTATACCCATATAGTAGAAGTGACGTTCGAATCAGTTGAAACT ATCCAAGATTACATCATACACCCTGCTCATGTTGGCTTTGGTGACGTCTACAGATCCTTCTGGGA AAAGTTGCTGATCTTCGATTACACTCCAAGAAAGAAATTGTCTGGTGGTGGTGGTTCTGGTGGTG GTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTATAATTTGGGAAACGCTTATTACAAACA GGGTGACTACCAAAAGGCCATCGAGTATTACCAAAAAGCTCTTGAACTGGACCCAAATAACGCT GAGGCATGGTATAATTTGGGTAACGCATACTATAAGCAAGGTGACTACCAAAAGGCAATTGAATA TTACCAAAAGGCCTTGGAGTTAGACCCTAATAACGCCGAAGCTTGGTACAATTTGGGTAATGCCT ACTATAAACAGGGTGACTATCAAAAGGCTATAGAGGACTACCAGAAAGCACTAGAACTTGATCCC AATAACTTGCAAGCAGAAGCCTGGAAGAATTTGGGTAATGCCTATTATAAGCAAGGTGACTATCA AAAAGCTATTGAATACTACCAAAAAGCTCTGGAATTGGACCCTAATAACGCTTCTGCTTGGTATAA TTTGGGTAATGCATACTACAAGCAAGGTGACTACCAGAAGGCAATAGAGTATTACCAAAAAGCCT TAGAACTAGACCCAAATAACGCCAAGGCTTGGTACAGAAGGGGTAATGCCTACTACAAGCAGGG TGACTACCAAAAAGCTATTGAGGACTACCAAAAAGCACTTGAACTGGATCCTAATAACAGATCTA GATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCCGGTCAAG ATAGAAGTGAAGCCACATTGATTAAAAGATTCAAAGGAGAAGGTGTTAGATACAAGGCTAAGCTG ATCGGTATCGATGAAGTTTCTGCTGCTAGAGGTGACAAATTGTGTCAAGACTCTATGATGAAGCT GAAGGGCGTTGTCGCAGGTGCCAGATCTAAGGGTGAACATAAGCAAAAGATATTTTTGACGATC TCATTCGGTGGTATTAAAATCTTCGATGAAAAGACTGGTGCTTTACAACATCACCATGCAGTACA CGAAATCTCTTACATCGCTAAGGATATCACAGACCATAGAGCATTCGGTTACGTTTGCGGTAAAG AAGGCAATCATAGATTTGTCGCTATTAAAACCGCCCAAGCCGCTGAACCAGTCATCTTGGATTTG AGAGACTTATTCCAGCTAATCTATGAACTAAAGCAAAGAGAAGAATTGGAAAAGAAAGCTGGTAG

CGCAGGATCGGCAGCCGGTAGCGGAGAATTTGGTTCTGCTGAGGCTGCAGCCAAAGAAGCTGC AGCCAAGGCCGGCTCTGCTGGTTCAGCTGCAGGCTCTGGTGAATTTGGTTCTGGTTCTCATATG GGTTCTCAATTTTGGGTAACTTCTCAAAAGACTGAAGCTTCCGAGAGATGTGGTTTGCAAGGCTC CTATATTTTAAGGGTGGAAGCCGAGAAGCTTACCCTACTTACGCTGGGTGCACAGAGTCAAATAT TGGAACCCCTGTTGTTCTGGCCATATACTTTATTGAGAAGATACGGTAGAGATAAAGTTATGTTC AGTTTCGAAGCTGGTAGAAGATGCCCAAGCGGTCCTGGAACTTTTACATTCCAGACATCACAAG GCAATGATATCTTTCAGGCAGTTGAAGCCGCTATTCAACAGCAAAAAGCCCAGGGTAAAGTCGG ACAGGCTCAAGACATTCTAAGATTGGAACACCATCACCATCATCATGGTTCTGGTGCTACTAACT TCTCTTTGTTGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGGGTTTGTCTTCAGTT TGTACATTCTCTTTCCAAACGAACTACCATACTTTGCTGAACCCTCACAACAACAATCCCAAAACT TCTTTGCTTTGCTACAGACATCCAAAAACCCCTATTAAGTATAGCTACAACAATTTCCCATCGAAA CATTGTAGTACTAAGAGCTTCCATTTGCAAAATAAGTGCTCCGAATCTTTGTCTATCGCTAAGAAC TCAATTAGAGCTGCAACTACAAATCAGACGGAACCACCTGAGTCGGATAATCACTCTGTAGCCAC CAAAATTTTGAACTTTGGTAAAGCTTGTTGGAAGCTGCAAAGACCATACACAATAATAGCCTTCAC CTCCTGTGCTTGCGGTTTGTTTGGTAAAGAACTGTTGCATAACACAAATTTGATTTCGTGGTCTTT GATGTTCAAGGCATTTTTCTTTTTGGTTGCAATCCTTTGCATCGCCTCTTTTACCACGACTATTAAT CAAATCTATGATTTGCACATCGACAGAATTAATAAGCCCGATTTGCCACTAGCTTCAGGTGAAAT CTCCGTTAATACTGCATGGATTATGTCAATCATTGTCGCCTTGTTCGGTTTAATCATCACAATTAA AATGAAAGGTGGTCCATTGTACATCTTCGGCTACTGTTTCGGTATATTCGGTGGTATAGTATATTC CGTTCCACCTTTTAGATGGAAACAAAACCCCAGTACCGCTTTCTTACTAAATTTCTTGGCACATAT CATCACAAACTTCACCTTCTACTACGCTTCTAGAGCTGCTTTGGGTTTGCCATTCGAATTAAGACC ATCTTTTACATTTTTGCTGGCTTTTATGAAATCGATGGGTTCTGCATTGGCCTTGATTAAAGATGC ATCTGACGTTGAAGGTGACACAAAATTCGGCATCAGTACCTTGGCTAGCAAGTACGGTTCTAGAA ATTTGACTTTGTTTTGTTCAGGTATCGTATTGTTATCCTACGTGGCAGCCATTTTAGCCGGTATCA TTTGGCCACAAGCTTTTAACAGTAATGTCATGCTACTTAGCCACGCAATATTGGCCTTCTGGCTG ATCTTGCAGACGAGAGATTTTGCTTTAACTAATTATGACCCTGAGGCAGGTAGAAGATTCTACGA ATTCATGTGGAAGCTGTACTACGCTGAATATTTGGTTTACGTCTTTATTAAGTTGTCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCTGAAGCATGGTACAACTTAGGCAAC GCATACTACAAGCAGGGTGACTACCAGAAGGCAATTGAGTATTACCAAAAAGCCTTAGAACTAGA CCCAAACAATGCCGAGGCTTGGTATAACTTGGGCAATGCTTATTACAAACAGGGTGACTATCAAA AGGCTATAGAATATTACCAAAAGGCACTTGAGCTGGACCCTAACAATGCAGAAGCCTGGTATAAC TTAGGCAATGCTTATTACAAGCAGGGTGACTATCAGAAGGCCATCGAGGACTACCAAAAGGCTT TGGAACTGGATCCAAACAATTTGCAGGCTGAAGCATGGAAGAATTTGGGTAACGCTTACTATAAA CAGGGTGACTATCAGAAAGCAATAGAATACTACCAAAAAGCCCTAGAACTTGACCCTAACAATGC CTCTGCTTGGTACAACTTGGGTAATGCTTACTATAAGCAGGGTGACTACCAAAAAGCTATCGAAT ATTACCAAAAAGCACTGGAATTGGACCCAAACAATGCAAAGGCCTGGTATAGAAGAGGTAACGC CTACTACAAACAGGGTGACTACCAAAAGGCTATTGAAGATTACCAAAAGGCTCTGGAACTAGATC CTAACAACAGATCTAGATCCGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTG GTGCTTCTGCAGAATACGTTAGAGCTCTGTTCGATTTCAACGGTAACGATGAAGAGGACTTGCCT TTTAAGAAAGGTGACATTTTGAGAATCAGGGACAAACCAGAAGAGCAATGGTGGAATGCTGAAG ATTCTGAGGGTAAAAGAGGAATGATTCCTGTTCCCTATGTCGAAAAGTACGGCTCAGCAGGTTC CGCTGCAGGATCTGGCGAATTCGGTTCAGCCGAGGCCGCTGCAAAAGAAGCCGCTGCAAAGGC TGGAAGTGCAGGCAGCGCCGCTGGTTCCGGAGAATTTGGTAGTTTGATTAAACATATGAGAGCC GAAGCTTTATTCGATTTTACTGGTAACTCCAAACTTGAACTGAATTTCAAGGCAGGTGACGTTATT TTCTTGTTGAGTAGAATTAATAAGGACTGGTTGGAAGGTACTGTTAGAGGTGCTACTGGAATATT CCCACTTTCTTTTGTGAAAATCCTGAAGGGCTCAGGTGCTACTAACTTCTCTTTGTTGAAGCAAG CAGGTGACGTTGAAGAAAATCCAGGTCCAATGAAATGTAGCACTTTTTCTTTCTGGTTCGTTTGC AAGATCATTTTCTTTTTCTTTTCTTTTAATATCCAAACTTCGATCGCAAATCCAAGAGAAAACTTCT TAAAGTGTTTCTCACAATACATTCCTAATAACGCCACGAATTTGAAGCTGGTATACACTCAGAACA ACCCACTGTACATGAGCGTGCTAAACTCGACAATCCATAATTTGAGATTCACTTCCGATACTACA CCCAAACCATTAGTAATCGTGACACCTTCTCATGTTTCACACATTCAAGGAACCATACTATGCTCT AAGAAAGTCGGTTTGCAGATTAGAACAAGGTCTGGTGGTCATGATAGTGAAGGCATGTCCTACA

TCAGTCAAGTTCCATTCGTTATCGTCGATTTGAGAAACATGAGGTCTATCAAAATAGACGTTCACT CACAGACGGCTTGGGTCGAGGCAGGTGCCACTTTGGGAGAAGTTTACTACTGGGTCAACGAAA AGAATGAAAATTTGTCTCTTGCTGCAGGTTACTGTCCAACTGTCTGCGCTGGTGGTCATTTTGGT GGTGGTGGTTATGGACCTCTTATGAGAAACTACGGTTTGGCCGCTGATAATATCATTGACGCACA TTTGGTAAATGTGCACGGTAAAGTTCTAGATAGAAAGTCAATGGGTGAAGATTTGTTTTGGGCAT TGAGAGGTGGTGGTGCTGAATCCTTTGGTATAATCGTAGCTTGGAAAATTAGATTGGTTGCAGTC CCAAAGTCTACAATGTTCTCAGTTAAGAAAATTATGGAAATCCATGAGCTGGTAAAGTTGGTGAA TAAGTGGCAAAACATCGCTTACAAGTACGATAAGGACTTGCTGCTAATGACCCATTTCATCACGA GAAACATCACTGATAACCAGGGTAAAAATAAGACAGCAATACACACCTACTTCTCTTCAGTTTTCT TGGGTGGTGTTGATTCCTTAGTGGATTTGATGAATAAGAGTTTCCCTGAACTGGGTATTAAGAAA ACTGATTGTAGACAATTGAGCTGGATCGACACAATCATATTCTATAGTGGTGTTGTCAACTACGA TACTGACAACTTCAACAAAGAAATCCTTCTGGATAGAAGTGCCGGACAAAATGGCGCTTTCAAAA TTAAGTTGGACTACGTTAAAAAGCCTATACCCGAGTCAGTATTTGTGCAGATCCTTGAAAAACTG TATGAAGAGGATATTGGTGCTGGAATGTACGCATTATATCCATACGGTGGTATAATGGATGAAAT CTCCGAGAGTGCCATACCATTCCCTCATAGAGCTGGTATCTTGTACGAACTGTGGTACATATGTT CTTGGGAAAAACAAGAGGATAACGAAAAGCACTTAAACTGGATCAGGAACATCTATAACTTCATG ACTCCTTACGTTTCTAAAAACCCCAGATTGGCTTATTTGAATTACAGAGATTTGGACATAGGTATC AACGATCCTAAAAATCCAAACAACTACACACAAGCAAGAATTTGGGGTGAAAAGTACTTCGGTAA AAATTTCGATAGATTGGTTAAAGTCAAGACCTTAGTTGACCCCAACAACTTTTTCAGAAACGAACA ATCTATTCCACCTTTGCCTAGACATAGGCACGGCTCTGGTGCTACTAACTTCTCTTTGTTGAAGC AAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGAACTGTAGCACTTTTTCTTTTTGGTTCGTTT GCAAGATAATATTTTTCTTTTTGTCCTTTAATATCCAAATCAGTATCGCCAACCCACAGGAAAACT TTTTAAAGTGTTTCTCTGAGTACATCCCCAACAACCCAGCTAACCCTAAGTTTATATATACACAAC ATGATCAGCTGTACATGAGCGTATTGAACTCGACCATTCAAAATTTGAGATTCACTTCTGACACTA CACCTAAGCCCTTGGTCATAGTAACTCCTTCTAATGTCTCACATATACAAGCTTCTATCTTGTGCT CTAAGAAAGTTGGTTTGCAGATTAGAACAAGGTCTGGTGGTCACGATGCAGAAGGTTTATCCTAT ATTAGTCAAGTCCCATTTGCCATAGTAGATTTGAGAAATATGCATACTGTGAAAGTTGACATACAC TCACAGACTGCTTGGGTGGAAGCAGGTGCCACATTGGGAGAGGTTTACTACTGGATCAACGAGA TGAACGAAAACTTTAGTTTCCCAGGTGGTTACTGTCCCACAGTCGGTGTTGGTGGTCATTTTTCT GGTGGTGGTTATGGAGCTTTAATGAGAAACTACGGTTTGGCTGCAGATAATATCATTGACGCACA TTTGGTGAACGTTGATGGTAAAGTTCTTGACAGAAAATCAATGGGTGAAGATTTGTTTTGGGCTA TCAGAGGTGGTGGTGGTGAAAATTTCGGTATAATCGCCGCTTGCAAAATTAAGTTGGTTGTCGTA CCTAGCAAAGCTACTATTTTCTCTGTCAAAAAGAACATGGAAATCCATGGTTTAGTAAAGTTGTTT AATAAGTGGCAAAACATCGCATACAAGTACGATAAGGATTTGATGCTTACCACGCATTTCAGAAC TAGGAACATCACAGATAACCATGGTAAAAATAAGACTACAGTTCACGGATACTTCTCTTCAATTTT CTTGGGTGGTGTTGATTCTCTTGTTGATTTGATGAATAAGTCATTCCCAGAACTGGGTATTAAAAA GACAGATTGTAAGGAACTGAGCTGGATCGACACCACGATTTTCTATAGTGGTGTGGTTAATTACA ACACCGCCAACTTCAAAAAGGAAATCTTGCTGGATAGATCCGCTGGTAAAAAGACCGCTTTTTCT ATTAAACTTGACTACGTTAAGAAACTGATCCCTGAAACTGCAATGGTTAAGATATTGGAGAAGCT GTACGAAGAGGAAGTCGGCGTAGGCATGTACGTTTTGTATCCATACGGTGGTATAATGGATGAG ATCTCCGAAAGTGCCATACCATTTCCTCATAGAGCTGGTATCATGTATGAATTATGGTACACCGC TACGTGGGAGAAGCAAGAAGATAACGAGAAACACATAAACTGGGTCAGATCTGTATACAACTTCA CTACACCTTACGTTTCTCAGAACCCAAGATTGGCATATTTGAACTACAGAGATTTGGACTTGGGT AAAACCAACCCCGAATCTCCAAATAACTATACGCAAGCAAGAATTTGGGGTGAAAAGTACTTCGG TAAAAATTTCAACAGATTGGTGAAGGTTAAGACAAAAGCCGATCCAAACAACTTCTTTAGAAACG AACAATCTATTCCACCATTGCCACCAAGACATCATGGTTCCGGCGCTACTAACTTCTCTTTGTTGA AGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGTCAGAAGAGTCCTTATTTGAATCTTCA CCACAAAAGATGGAGTACGAAATCACTAACTACTCTGAGAGACATACAGAATTGCCTGGACACTT CATCGGTTTGAACACAGTTGACAAGCTGGAAGAGTCTCCATTGAGAGATTTCGTCAAGTCCCATG GTGGTCACACCGTAATTAGTAAGATCTTGATAGCTAACAACGGTATCGCTGCAGTCAAGGAAATT AGATCTGTTAGAAAGTGGGCATATGAAACCTTTGGTGACGATAGAACGGTCCAATTCGTAGCTAT GGCAACTCCTGAAGACTTGGAGGCCAATGCTGAATATATCAGAATGGCCGATCAATACATTGAA

GTTCCAGGTGGTACAAATAACAATAACTACGCTAATGTCGACTTAATAGTAGATATCGCTGAAAG AGCAGACGTGGATGCCGTTTGGGCTGGTTGGGGACATGCTTCCGAAAACCCTTTGTTACCCGAA AAATTGTCTCAGAGTAAGAGAAAAGTTATTTTTATTGGTCCACCTGGAAATGCAATGAGATCATTA GGTGACAAGATATCCAGTACTATCGTGGCACAATCAGCCAAAGTTCCATGTATTCCTTGGTCCGG CACCGGTGTTGACACGGTGCATGTTGATGAAAAGACTGGTTTGGTTTCTGTAGATGACGATATCT ATCAGAAGGGATGTTGCACTTCACCTGAAGATGGTTTGCAAAAGGCTAAGAGAATCGGTTTCCC AGTTATGATCAAGGCATCAGAAGGTGGTGGTGGTAAAGGTATCAGGCAGGTCGAAAGAGAAGA GGATTTCATCGCTCTGTACCATCAAGCCGCTAATGAAATACCCGGTTCTCCAATTTTCATAATGAA ACTAGCTGGAAGGGCAAGACATTTGGAAGTTCAGCTACTTGCTGACCAATACGGCACTAATATTT CCTTGTTCGGTAGAGATTGCAGTGTTCAAAGAAGACATCAAAAGATTATCGAAGAGGCACCAGTC ACTATAGCAAAAGCCGAAACATTTCACGAGATGGAAAAGGCAGCTGTTAGATTGGGTAAATTGGT CGGATATGTAAGTGCTGGAACAGTCGAATATTTGTACAGCCATGACGATGGTAAATTCTACTTTT TGGAACTTAACCCAAGATTACAAGTTGAGCACCCTACTACAGAAATGGTTTCTGGTGTTAATTTG CCAGCTGCACAACTGCAGATTGCTATGGGTATCCCTATGCATAGAATCAGTGATATCAGGACTCT GTACGGTATGAATCCACACAGCGCTTCGGAGATTGACTTCGAATTCAAAACTCAGGATGCAACTA AGAAACAAAGAAGACCAATCCCAAAGGGTCATTGTACCGCTTGCAGAATTACGTCCGAAGACCC CAATGATGGTTTTAAACCATCTGGTGGTACTTTGCACGAACTAAACTTTAGAAGCTCGTCTAATGT CTGGGGTTATTTCTCAGTAGGCAACAACGGTAACATCCATTCTTTTTCAGATTCCCAGTTCGGTC ACATCTTCGCATTTGGAGAAAATAGGCAAGCCTCTAGAAAGCATATGGTTGTCGCTCTTAAAGAA CTGTCAATCAGAGGTGACTTCAGAACCACGGTTGAATACTTAATTAAACTGTTGGAAACTGAAGA CTTCGAAGATAATACGATTACTACAGGTTGGTTGGACGATTTGATAACCCATAAGATGACGGCAG AAAAACCTGATCCCACCTTGGCCGTTATCTGTGGTGCCGCTACGAAGGCCTTTTTAGCTTCTGAA GAGGCTAGACATAAGTACATAGAAAGCCTGCAAAAGGGTCAGGTACTATCGAAAGACTTACTAC AAACAATGTTTCCTGTGGATTTCATCCACGAAGGTAAAAGATACAAGTTTACTGTTGCTAAGTCTG GCAACGATAGGTACACGTTGTTCATTAATGGTAGCAAGTGCGACATCATTCTAAGACAACTTTCA GATGGTGGTTTGCTGATCGCAATTGGTGGTAAATCACATACTATCTATTGGAAGGAAGAGGTCG CAGCCACAAGATTGAGTGTAGACAGCATGACCACGTTGTTAGAGGTTGAAAACGATCCAACTCA ATTAAGAACACCATCTCCTGGTAAACTTGTGAAATTTCTGGTTGAAAATGGCGAGCATATAATCAA GGGTCAACCCTACGCTGAGATTGAAGTTATGAAAATGCAGATGCCATTGGTTTCTCAAGAAAACG GTATAGTTCAACTACTTAAACAGCCTGGATCAACCATAGTAGCTGGTGACATCATGGCAATTATG ACGTTAGACGATCCATCCAAGGTGAAACATGCTCTTCCTTTTGAGGGTATGCTGCCCGATTTCGG TTCTCCAGTTATTGAAGGCACTAAACCAGCATACAAGTTTAAATCGTTGGTTTCTACACTGGAAAA CATCCTAAAGGGTTACGATAACCAAGTTATTATGAATGCTTCTTTGCAACAGTTGATAGAAGTCTT GAGAAATCCTAAGTTACCCTATTCAGAATGGAAATTGCATATTAGCGCTCTTCACTCCAGATTGC CTGCAAAATTGGATGAACAAATGGAAGAGCTAGTCGCTAGATCTTTGAGAAGAGGTGCTGTATTT CCAGCAAGGCAATTGAGTAAGCTAATTGACATGGCAGTTAAAAACCCAGAATACAACCCTGATAA ACTGTTGGGTGCCGTAGTGGAACCATTGGCAGATATTGCCCATAAGTACTCTAATGGTTTAGAAG CTCATGAGCACTCAATCTTCGTGCATTTCTTGGAAGAGTACTACGAGGTTGAAAAATTGTTCAAC GGTCCTAACGTCAGAGAAGAGAACATCATCCTGAAGTTGAGAGATGAAAACCCAAAGGACTTGG ATAAAGTCGCTCTTACTGTACTGAGTCATAGCAAGGTTTCTGCCAAAAATAACTTAATCCTAGCTA TCCTGAAGCACTACCAACCTTTGTGTAAGCTGTCATCCAAAGTTTCTGCAATATTTTCAACTCCAT TGCAACATATCGTAGAGCTTGAATCTAAGGCTACCGCAAAAGTGGCTTTGCAGGCAAGAGAAATT TTGATCCAAGGTGCTTTGCCATCAGTTAAAGAAAGAACAGAGCAAATAGAACACATCCTGAAGAG TAGCGTTGTCAAAGTCGCATACGGTTCGTCTAATCCTAAGAGATCTGAACCCGATTTGAATATAC TTAAGGATTTGATCGATTCAAATTACGTAGTGTTTGACGTTTTACTACAGTTCTTAACTCATCAAGA TCCTGTTGTCACAGCTGCAGCCGCTCAAGTCTATATAAGAAGGGCCTATAGAGCTTACACTATCG GTGACATTAGGGTACACGAAGGCGTGACAGTTCCAATCGTGGAATGGAAATTTCAATTGCCCTC CGCAGCCTTTAGTACCTTCCCAACGGTAAAGTCAAAAATGGGTATGAACAGAGCTGTTTCTGTTT CTGATTTGAGCTATGTGGCTAATTCGCAATCATCCCCTTTAAGAGAAGGTATTCTAATGGCTGTG GACCATTTGGACGATGTTGATGAAATTTTGTCTCAATCTTTGGAAGTTATTCCAAGACACCAAAGT AGCTCGAATGGTCCCGCTCCAGATAGGTCTGGATCTTCAGCAAGTTTAAGCAACGTAGCCAATG TGTGTGTTGCTTCCACTGAGGGTTTTGAAAGTGAAGAGGAAATCTTGGTTAGATTGAGAGAAATT

TTGGATTTGAACAAACAAGAATTGATTAATGCTTCCATCAGAAGGATCACATTCATGTTCGGTTTT AAAGATGGTAGTTACCCTAAGTACTACACCTTTAATGGTCCCAACTACAACGAGAACGAAACTAT CAGACATATCGAACCTGCCTTAGCTTTCCAATTGGAACTGGGTAGATTGTCAAACTTCAACATCA AGCCAATTTTCACTGATAACAGAAACATCCATGTGTACGAAGCTGTTTCAAAGACATCCCCATTA GATAAGAGATTTTTCACCAGAGGCATCATTAGGACGGGTCACATTAGAGATGATATTAGCATACA AGAGTACTTGACTTCGGAAGCTAACAGATTAATGTCTGACATCCTAGATAATTTGGAAGTTACCG ACACGTCGAACTCTGATTTGAACCATATCTTTATTAACTTCATCGCAGTGTTCGACATATCTCCTG AGGATGTTGAAGCTGCATTTGGTGGTTTCTTGGAAAGATTCGGTAAAAGATTGCTGAGATTGAGA GTCTCCAGTGCTGAAATCAGAATCATCATTAAGGATCCACAAACTGGTGCCCCTGTACCCCTGA GAGCTTTGATCAATAATGTTTCTGGTTACGTAATTAAAACCGAGATGTACACGGAAGTCAAGAAT GCTAAGGGTGAATGGGTATTCAAGAGCTTGGGTAAACCCGGCTCGATGCACTTAAGACCAATTG CAACACCATATCCTGTCAAAGAATGGTTGCAACCTAAGAGATACAAAGCCCACTTAATGGGTACT ACATACGTTTACGATTTCCCAGAATTGTTCAGACAGGCTTCTTCTTCTCAATGGAAGAATTTTTCC GCCGACGTTAAGCTGACTGACGATTTCTTTATCAGTAACGAACTAATCGAGGATGAAAATGGTGA ACTTACAGAGGTTGAAAGAGAGCCAGGAGCAAATGCCATTGGCATGGTCGCTTTTAAGATCACT GTAAAGACACCAGAATATCCTAGGGGTAGACAATTCGTAGTGGTTGCAAACGACATCACCTTTAA AATTGGTTCTTTCGGACCTCAAGAAGATGAGTTTTTCAATAAGGTTACTGAATACGCTAGGAAAA GAGGTATACCAAGAATCTACTTGGCCGCTAATTCTGGAGCAAGGATTGGCATGGCCGAGGAAAT AGTGCCTTTATTTCAGGTTGCATGGAACGACGCAGCCAACCCAGATAAGGGATTCCAATATTTGT ATTTGACTTCTGAGGGTATGGAAACATTGAAAAAGTTCGATAAGGAAAACTCAGTGCTGACCGAG AGAACTGTTATTAATGGAGAGGAAAGGTTCGTAATCAAAACTATAATCGGTTCTGAAGATGGTTT GGGCGTGGAGTGTCTGAGAGGTAGCGGTTTGATTGCTGGTGCAACTTCTAGAGCTTACCATGAT ATTTTTACTATCACACTGGTCACTTGCAGATCTGTAGGCATAGGTGCTTATTTGGTTAGATTGGGT CAAAGGGCCATCCAGGTCGAAGGCCAACCTATTATATTGACTGGTGCCCCCGCTATAAACAAAA TGCTGGGTAGAGAAGTTTATACCTCCAATTTGCAGTTGGGTGGTACGCAAATCATGTACAATAAC GGTGTTTCTCATTTGACAGCTGTAGACGATTTGGCTGGTGTGGAAAAGATTGTTGAATGGATGTC ATATGTGCCAGCTAAAAGAAACATGCCCGTTCCAATATTGGAAACTAAGGACACATGGGATAGAC CAGTAGATTTTACCCCTACGAATGACGAAACCTATGATGTGAGATGGATGATTGAGGGTAGGGA AACTGAGTCTGGTTTTGAATACGGTTTGTTCGATAAGGGTTCTTTCTTTGAAACATTATCAGGCTG GGCCAAGGGTGTCGTAGTGGGAAGAGCTAGATTGGGTGGTATTCCTCTAGGTGTTATTGGTGTA GAAACTAGAACAGTTGAAAATTTGATCCCCGCAGATCCAGCCAACCCTAATTCTGCTGAAACTTT AATTCAGGAACCTGGTCAAGTTTGGCATCCCAACTCAGCTTTTAAAACCGCACAGGCCATTAATG ATTTCAACAACGGTGAACAATTGCCAATGATGATACTGGCTAACTGGAGAGGTTTTTCTGGTGGT CAAAGGGATATGTTCAACGAAGTTTTGAAGTACGGTAGTTTTATCGTCGACGCACTGGTAGATTA CAAGCAACCTATCATAATATACATTCCACCAACTGGTGAATTAAGAGGTGGTTCTTGGGTTGTCG TAGACCCAACCATTAACGCAGATCAGATGGAAATGTACGCCGATGTGAATGCTAGAGCAGGTGT TTTGGAACCACAAGGAATGGTTGGTATTAAGTTTAGAAGAGAAAAATTGCTGGATACTATGAACA GATTAGACGATAAGTACAGGGAATTGAGATCTCAACTGAGCAATAAGTCTTTGGCTCCAGAAGTT CATCAACAGATCTCTAAGCAACTGGCTGATAGGGAAAGAGAATTGTTGCCAATATACGGTCAGAT CTCATTGCAATTTGCCGACTTACACGATAGGTCATCCAGAATGGTGGCTAAGGGTGTTATTTCAA AAGAATTAGAGTGGACAGAAGCTAGAAGATTTTTCTTTTGGAGATTGAGAAGAAGATTGAACGAG GAATATTTGATTAAAAGATTGTCACATCAAGTTGGCGAGGCTTCTAGATTGGAAAAGATCGCAAG GATTAGATCTTGGTATCCAGCATCAGTCGATCACGAAGACGATAGACAAGTAGCCACTTGGATTG AGGAAAATTACAAGACACTGGACGATAAGTTGAAGGGTTTAAAGCTAGAATCCTTTGCCCAAGAC TTGGCTAAAAAGATTAGAAGTGACCATGATAATGCTATCGATGGTTTGAGTGAAGTTATTAAAATG CTTAGCACTGACGATAAGGAAAAACTGTTGAAGACATTGAAGAAACTGTCTGGTGGTGGTGGTT CTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCTTGGTATAACTTGGGAAATGCTTA TTACAAGCAGGGTGACTACCAAAAGGCCATAGAATACTACCAAAAGGCTCTTGAGCTGGATCCT AATAACGCAGAAGCCTGGTATAACTTAGGCAATGCATACTATAAACAAGGTGACTACCAAAAGGC AATAGAGTACTACCAAAAGGCCTTGGAATTAGATCCAAATAACGCTGAGGCATGGTATAACTTGG GCAACGCCTACTATAAACAGGGTGACTATCAAAAGGCTATAGAAGATTACCAGAAGGCACTAGA GCTTGATCCTAATAACTTGCAAGCCGAAGCTTGGAAGAACTTAGGAAATGCATACTATAAGCAAG

GTGACTATCAAAAAGCTATTGAATATTACCAAAAGGCTCTGGAGTTGGATCCAAATAACGCATCT GCTTGGTACAACTTAGGCAACGCCTACTATAAGCAGGGTGACTATCAAAAAGCAATTGAATATTA TCAAAAGGCCTTAGAGCTAGATCCTAATAACGCTAAAGCATGGTATAGGAGAGGCAATGCATACT ACAAACAGGGTGACTACCAAAAAGCTATAGAAGATTACCAAAAGGCACTTGAACTGGATCCAAAT AACAGATCTAGATCCGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTT CTGGTTCTCATATGAGATTGGGAGCCCAATCTATTCAGCCAACCGCTAACTTAGATAGAACGGAC GATTTGGTCTATTTGAATGTAATGGAATTGGTTAGAGCTGTTTTGGAGTTGAAAAATGAACTAGCA CAATTGCCACCAGAAGGTTACGTGGTTGTCGTAAAGAATGTTGGTTTGACTCTTAGAAAGTTGAT AGGCTCGGTCGACGATTTGCTACCATCTTTGCCATCTTCTTCTAGAACTGAAATAGAGGGTACAC AAAAGCTTCTGAACAAAGATTTGGCTGAATTGATTAATAAGATGAGATTGGCACAACAGAACGCC GTTACTTCTTTGTCTGAGGAGTGTAAGAGACAAATGCTAACTGCTTCTCATACTTTGGCTGTTGAT GCAAAGAACTTGTTAGACGCTGTGGATCAAGCAAAAGTTTTAGCCAATTTGGCTCACCCACCTGC CGAAGGTTCTGCTGGATCAGCTGCAGGATCCGGCGAATTTGGTTCTGCTGAAGCCGCTGCAAAA GAGGCTGCTGCAAAAGCTGGATCTGCAGGTAGTGCTGCTGGTAGCGGAGAATTTGGTTCTGGT GCCATGGCTACTCCTGGTTCAGAAAACGTTCTACCAAGAGAACCATTGATTGCAACAGCCGTGA AGTTCTTGCAGAACTCTAGAGTTAGACAATCTCCATTGGCAACTAGAAGAGCATTTTTGAAAAAG AAAGGTTTGACCGACGAGGAAATTGATATGGCTTTCCAACAGTCCGGTACTGCAGCCGATGAAC CATCTTCATTGTGGGGAAGTGGC

Figure 15D

Complete SCF Gene Cassette Nucleotide Sequence

ATGGGTTCTGCTGGTTCAGCTGCAGGTTCTGGTGAATTCGGTTCCGCTGGTAGTGCCGCTGGTT CTGGTGAATTTGGTTCTGCTGGTTCAGCAGCCGGTTCTGGTGAATTCTCCTATTACCATCACCAT CACCATCACTTGGAATCTACTTCATTATACAAAAAGGCTGGTTCCGGTAGTGCCAGAAACGCTTA CTTGAGAAAGAAAATTGCTAGATTGAAGAAAGATAATTTGCAATTGGAAAGAGATGAACAAAACTT GGAAAAGATTATCGCTAATTTGAGAGATGAAATAGCAAGATTGGAAAATGAAGTTGCTTCTCATG AACAAGGTTCCGCAGGTAGTGCCGCCGGTTCTGGTGAATTTGCTGAAGCCGCTGCAAAGGAAG CCGCTGCAAAAGCAGGTTCTGCCGGTTCAGCCGCTGGTAGTGGTGAATTTTCTTACTATCACCAT CACCATCATCACTTGGAATCTACCTCATTATATAAAAAGGCCGGTTCCGGTAGTAACTTGGTTGC TCAATTAGAAAATGAAGTCGCATCATTGGAAAACGAAAACGAAACTTTGAAAAAGAAAAACTTACA TAAGAAAGATTTGATCGCTTACTTAGAAAAGGAAATAGCAAATTTGAGAAAGAAAATAGAAGAAG GTTCCGCTGGTAGTGCAGCCGGTAGTGGTGAATTCGGTTCTGCTGAAGCTGCAGCCAAGGAAG CTGCAGCCAAAGAAGCCGCTGCTAAAGAAGCTGCAGCCAAAGCTGGTTCTGCAGGTTCTGCCG CAGGTTCCGGTGAATTTGGTTCTTCATACTATCACCATCACCACCACCACTTGGAATCTACCTCA TTATACAAGAAAGCTGGTTCCGGTAGTCAAAAGGTCGCTGAATTGAAAAACAGAGTAGCTGTTAA GTTGAACAGAAACGAACAATTGAAAAATAAGGTAGAAGAATTGAAAAATAGAAACGCTTACTTGA AAAACGAATTGGCAACTTTGGAAAACGAAGTAGCTAGATTAGAAAACGATGTTGCTGAAGGTTCT GCTGGTTCTGCTGCTGGTTCTGGTGAATTCGCTGAAGCAGCCGCTAAGGAAGCAGCCGCTAAA GCCGGTTCCGCCGGTTCTGCTGCGGGCTCTGGTGAATTTTCCTACTATCACCATCATCATCACC ACTTGGAATCTACATCATTATATAAGAAAGCCGGTTCCGGTAGTAATGAAGTTACTACATTGGAAA ACGATGCTGCTTTTATTGAAAACGAAAACGCATACTTAGAAAAGGAAATCGCTAGATTGAGAAAG GAAAAGGCCGCTTTGAGAAATAGATTAGCTCATAAGAAAGGTTCTGCTGGTAGCGCTGCTGGCT CTGGTGAATTTGGTTCCGCCGAAGCCGCTGCTAAGGAAGCCGCTGCCAAAGAAGCCGCTGCCA AGGAAGCCGCTGCTAAGGCTGGTTCCGCCGGTTCAGCTGCAGGCTCTGGTGAATTCGGTTCTA GACCACCTACCATCTCTAATCCACCTCCATTGATTTCCAGTGCTAAACATCCATCCGTCGGTAGT GCAGGTTCCGCTGCCGGCTCTGGCGAATTTGCCGAAGCTGCTGCCAAAGAAGCAGCCGCTAAA GCTGGTTCAGCAGGTTCCGCTGCCGGATCTGGCGAATTCAATTTCTTGCAATCTAGACCAGAAC CTACTGCTCCTCCAGAAGAAAGTTTCAGATCTGGTGGTTCAGCTGGTTCCGCCGCAGGATCTGG CGAATTTGGTTCCGCAGAAGCTGCCGCTAAAGAAGCTGCTGCAAAAGAAGCAGCCGCCAAAGAA GCTGCTGCAAAAGCCGGTAGTGCTGGTTCAGCTGCCGGTTCCGGTGAATTCGGTTCTTCAAAAG GTACCGGTTTAAATCCAAACGCTAAAGTTTGGCAAGAAATTGCTCCTGGTAACGGTTCTGCAGGT TCCGCAGCTGGTTCCGGTGAATTCGCCGAGGCCGCTGCTAAGGAAGCAGCAGCCAAAGCAGGT

AGTGCTGGTTCCGCAGCTGGTTCAGGTGAATTCCCAGACGGTGGTACCACTTTCGAACATTTGT GGTCCAGTTTAGAACCTGATTCTACATACGGTTCTGCCGGTTCTGCAGCAGGCAGCGGTGAATT CGGTTCTGCCGAAGCTGCTGCTAAAGAAGCTGCTGCCAAGGAAGCTGCTGCTAAGGAAGCTGC TGCCAAAGCCGGTAGTGCAGGTTCTGCTGCCGGTTCAGGTGAATTTGGTTCTTCTTACTATCACC ACCACCACCATCACTTGGAATCTACATCATTATACAAGAAAGCCGGTTCTGGTAGTAAGAGAATC GCATACTTAAGAAAGAAAATCGCTGCATTGAAGAAAGATAACGCAAACTTAGAAAAGGACATCGC TAACTTGGAAAACGAAATCGAAAGATTGATTAAAGAAATCAAAACCTTGGAAAATGAAGTTGCATC TCATGAACAAGGTTCAGCCGGTTCTGCAGCGGGCTCCGGTGAATTTGCCGAAGCTGCAGCAAAA GAAGCTGCCGCTAAGGCTGGTAGTGCTGGTTCTGCTGCAGGCAGCGGTGAATTTTCTTACTACC ACCATCACCACCATCACTTGGAATCTACTTCATTATATAAGAAAGCAGGTTCTGGTAGTAACTTGT TAGCAACATTAAGATCTACCGCTGCAGTCTTGGAAAACGAAAACCATGTATTGGAAAAAGAAAAG GAAAAATTGAGAAAGGAAAAAGAACAATTGTTGAATAAGTTGGAAGCTTACAAAGGTTCAGCAGG TTCTGCAGCGGGCTCTGGCGAATTCGGTTCCGCCGAAGCTGCAGCAAAGGAAGCTGCAGCTAA AGAGGCCGCTGCAAAAGAAGCTGCTGCCAAAGCAGGTAGTGCAGGTTCCGCAGCCGGCTCCG GCGAATTTGGTTCACCAGCTACATCCCAACATCCTCCACCTCCACCTGGTCATAGATCTCAAGCT CCTTCACATGGTTCCGCAGGTAGTGCAGCTGGATCTGGCGAATTCGCCGAAGCTGCCGCTAAG GAAGCTGCTGCAAAAGCTGGTTCCGCTGGTTCAGCAGCAGGTTCCGGTGAATTCGAATTGAATT CTTTGTTGATATTGTTAGAAGCAGCCGAATATTTGGAAAGAAGAGATAGAGGTTCTGCCGGTAGT GCTGCAGGTAGCGGCGAATTTGGTTCTGCAGAAGCAGCCGCCAAGGAAGCAGCTGCAAAAGAA GCAGCAGCTAAAGAAGCAGCTGCAAAAGCCGGTTCTGCTGGTTCAGCCGCAGGATCTGGAGAA TTCGGTTCCAGACCACCTACAATTTCCAATCCACCTCCATTGATCTCTTCTGCCAAGCATCCATC CGTTGGTAGTGCAGGTTCAGCTGCCGGTAGTGGTGAATTTGCCGAAGCCGCCGCTAAGGAAGC CGCCGCCAAAGCAGGTTCAGCCGGTTCCGCCGCAGGTTCAGGTGAATTCAATTTCTTGCAGTCA AGACCAGAACCTACCGCTCCTCCAGAGGAGAGTTTCAGATCTGGTGGTAGTGCCGGTTCAGCTG CCGGCTCTGGAGAATTTGGTTCTGCAGAGGCTGCTGCCAAGGAAGCCGCAGCTAAAGAAGCCG CTGCGAAAGAAGCCGCCGCTAAAGCTGGTAGTGCAGGTAGTGCTGCGGGATCTGGCGAATTCG GTTCTTCTAAGGGTACTGGTTTGAACCCTAATGCCAAGGTCTGGCAAGAAATCGCCCCTGGTAA CGGTTCCGCAGGTTCCGCCGCAGGTAGTGGTGAATTCGCCGAGGCTGCCGCCAAGGAAGCCG CCGCTAAGGCAGGTAGTGCTGGTTCAGCGGCCGGCTCTGGTGAATTTCCAGACGGTGGTACAA CCTTTGAGCATTTGTGGTCCAGTTTAGAACCTGATTCTACGTACGGTTCTGCTGGTTCCGCTGCA GGATCTGGCGAATTCGGTTCCGCGGAAGCCGCCGCAAAAGAAGCCGCCGCCAAAGAAGCCGC CGCAAAGGAAGCCGCAGCAAAGGCAGGTAGTGCCGGCTCCGCCGCTGGCAGTGGCGAATTTG GTTCTTCATATTATCACCATCATCATCATCACTTGGAATCTACTTCATTATACAAGAAAGCAGGTT CCGGTTCTAAAAGAATTGCTTACTTAAGAAAGAAAATCGCGGCTTTGAAGAAAGACAATGCTAAC TTAGAAAAAGATATTGCCAACTTGGAAAATGAAATCGAAAGATTAATTAAGGAAATTAAAACATTG GAAAACGAAGTTGCATCACATGAACAAGGTTCAGCTGGTTCCGCTGCAGGGTCCGGCGAATTTG CAGAAGCCGCCGCCAAGGAAGCCGCAGCCAAAGCTGGTAGTGCAGGTTCTGCCGCTGGCTCTG GCGAATTTTCTTACTATCATCATCACCATCACCACTTGGAATCTACTTCATTATACAAGAAAGCGG GTTCAGGTTCTAACTTGTTAGCAACTTTAAGATCTACAGCCGCTGTTTTAGAAAATGAAAACCATG TCTTAGAAAAAGAAAAGGAAAAGTTGAGAAAGGAAAAGGAACAATTATTAAATAAGTTAGAAGCC TACAAGGGTTCAGCAGGTTCCGCAGCAGGCTCAGGCGAATTTGGTTCTGCAGAAGCGGCTGCT AAGGAAGCTGCCGCAAAGGAAGCAGCTGCTAAGGAGGCCGCTGCAAAGGCTGGTTCTGCTGGT TCCGCCGCGGGCTCTGGAGAATTCGGTTCCGCTTTGGTTGATGACGCCGCTGATTATGAACCTC CACCTTCAAATAACGAAGAAGCTTTAGGTTCCGCTGGTTCCGCTGCAGGTTCCGGCGAGTTCGC AGAAGCCGCAGCAAAAGAAGCCGCAGCTAAGGCAGGTAGTGCCGGATCCGCCGCTGGCAGTG GAGAATTCAGAGAATTGTTCGATGACCCATCTTACGTCAACGTACAAAATTTGGATAAAGCTAGA CAAGGTTCCGCCGGTTCTGCAGCGGGATCTGGGGAATTTGGTTCTGCAGAAGCTGCCGCCAAA GAAGCTGCAGCTAAAGAAGCCGCAGCCAAAGAAGCTGCTGCTAAGGCCGGTTCTGCTGGTTCT GCCGCAGGATCTGGGGAATTCGGTTCCAAGAATACTAAGAGTATGAACTTCGATAACCCAGTTTA CAGAAAGACTACAGAAGAAGAAGGTTCAGCCGGTTCAGCCGCCGGTTCCGGTGAATTTGCAGA GGCTGCCGCTAAAGAGGCTGCCGCTAAGGCCGGTAGTGCTGGTTCTGCAGCCGGCTCCGGAG AATTCAGATCTTTGCCATCCACATGGATTGAAAACAAATTATACGGCATGTCAGACCCTAATTGG

GGTTCTGCAGGTTCAGCTGCGGGATCTGGTGAATTCGGTTCAGCAGAAGCCGCAGCCAAGGAA GCCGCTGCAAAGGAGGCCGCTGCCAAAGAAGCAGCTGCTAAGGCTGGTTCAGCCGGTTCCGCA GCCGGCAGTGGTGAATTTGGTAGTGTTGTCGATAATTCTCCACCTCCAGCTTTGCCTCCAAAGAA AAGACAATCTGCTCCATCTGGTTCAGCAGGTTCAGCCGCTGGTTCAGGTGAATTTGCCGAAGCA GCTGCCAAGGAAGCTGCCGCCAAGGCGGGCAGTGCAGGTTCGGCTGCGGGGTCTGGTGAATT CACTCAAAGATCTAAACCACAACCTGCAGTTCCTCCAAGACCATCTGCTGACTTGATTTTAGGTT CCGCCGGTTCCGCAGCTGGCTCTGGCGAATTCGGTTCCGCTGAGGCTGCCGCTAAAGAAGCGG CCGCTAAAGAGGCAGCCGCTAAAGAGGCGGCCGCTAAAGCAGGTTCTGCAGGTTCAGCAGCAG GTAGTGGTGAATTTGGTTCTACAGATGAAGAAAGAGAAGAAACCGAAGAAGAAGTTTATTTGTTG AACTCTACCACTTTGGGTTCAGCTGGTTCTGCTGCGGGTTCTGGCGAATTTGCAGAAGCAGCTG CTAAGGAAGCCGCGGCAAAGGCTGGTTCTGCGGGCTCCGCCGCAGGTTCTGGTGAATTTGATG GTAATGTATCTGGTACTCAAAGATTAGACTCAGCTACCGTTAGAACTTATTCATGCGGTTCTGCC GGTAGTGCAGCGGGCTCTGGGGAATTCGGTTCCGCAGAAGCCGCTGCAAAAGAAGCCGCTGCA AAAGAAGCCGCTGCGAAGGAGGCTGCTGCTAAGGCAGGTTCCGCCGGTAGTGCTGCGGGTTCC GGCGAATTTGGTTCCAGTTACTATCACCATCATCACCACCACTTGGAATCCACAAGTTTATATAA GAAAGCTGGTTCTGGTTCACAAAAGGTAGCTCAATTGAAAAATAGAGTTGCATACAAGTTGAAGG AAAACGCTAAGTTGGAAAACATAGTAGCAAGATTAGAAAACGATAACGCTAATTTGGAAAAGGAC ATCGCAAATTTGGAAAAGGATATAGCTAACTTGGAAAGAGATGTTGCTAGAGGTTCTGCTGGTAG TGCCGCAGGCTCTGGCGAATTCGCTGAAGCTGCCGCTAAAGAGGCTGCGGCTAAAGCTGGTTC AGCTGGTTCTGCAGCGGGGTCTGGTGAATTTTCTTATTATCACCATCATCACCATCACTTGGAAT CCACCAGTTTATACAAGAAAGCCGGCTCTGGTTCAAACACTGTTAAGGAATTGAAAAATTACATT CAAGAATTGGAAGAAAGAAACGCTGAATTGAAAAATTTGAAGGAACATTTGAAGTTTGCAAAAGC CGAATTGGAATTCGAATTAGCAGCCCATAAATTTGAAGGTTCTGCCGGTTCTGCCGCCGGATCT GGAGAATTTGGTTCTGCGGAGGCTGCCGCTAAAGAAGCCGCCGCTAAAGAGGCTGCAGCTAAG GAAGCTGCAGCAAAGGCTGGTTCTGCCGGTTCCGCTGCCGGCTCCGGCGAATTTGGTTCACAT GATGACTCCTTGCCACATCCTCAACAAGCTACAGATGACTCTGGTCATGAATCCGACGGTTCCG CAGGCTCTGCTGCCGGCTCCGGCGAGTTTGCTGAAGCCGCTGCTAAAGAGGCTGCTGCTAAAG CCGGTTCTGCCGGTTCAGCAGCTGGATCTGGAGAATTTGGTTCCCCAAATGCTGGTAGTGTTGA ACAAACCCCAAAGAAACCTGGTTTGAGAAGAAGAGGTAGTGCTGGTTCTGCCGCTGGCTCCGGA GAATTTGGTTCAGCCGAAGCTGCGGCCAAAGAGGCTGCTGCAAAGGAGGCTGCGGCTAAGGAA GCCGCCGCTAAAGCCGGTTCAGCTGGTTCCGCGGCAGGCTCCGGGGAATTTGGTTCTTCTTATT ATCACCACCACCACCATCACTTGGAATCCACAAGTTTATACAAGAAAGCAGGCTCTGGTTCATTC GAAAACGTCACTCATGAATTCATTTTGGCAACCTTAGAAAACGAAAACGCTAAGTTGAGAAGATT AGAAGCAAAGTTGGAAAGAGAATTGGCTAGATTAAGAAATGAAGTAGCTTGGTTGGGTTCTGCG GGCTCGGCCGCTGGCTCTGGTGAATTCGCCGAAGCTGCGGCCAAGGAGGCTGCCGCAAAGGC CGGTTCTGCCGGTTCCGCAGCGGGATCCGGCGAATTTTCTTACTACCATCATCACCATCACCAC TTGGAATCCACAAGTTTATACAAGAAAGCGGGTTCTGGTTCACAAAAAGTTGAAGAATTGAAAAA TAAGATAGCAGAATTGGAAAACAGAAACGCTGTAAAGAAAAATAGAGTTGCACATTTGAAGCAAG AAATCGCTTACTTGAAGGATGAATTAGCAGCCCATGAATTCGAAGGTAGTGCCGGTTCCGCTGC TGGCTCAGGCGAATTTGGTAGTGCAGAAGCTGCCGCTAAGGAGGCTGCCGCCAAAGAAGCAGC CGCAAAAGAAGCTGCCGCAAAAGCCGGTTCTGCGGGCTCTGCTGCCGGATCCGGCGAATTCGG TTCAGTCTCCAGTACTAAATTAGTATCCTTTCATGATGACAGTGATGAAGACTTGTTACATATCGG TTCTGCAGGCTCAGCCGCTGGCTCTGGAGAGTTTGCAGAGGCAGCTGCTAAAGAAGCCGCCGC AAAGGCAGGTTCTGCAGGTTCTGCAGCTGGTAGTGGTGAATTCGCTGCTGCAACCCCAATATCT ACTTTTCATGATGACTCAGACGAAGACTTGTTGCATGTCGGTTCCGCAGGTTCAGCAGCGGGAT CCGGTGAATTTGGTTCAGCAGAAGCTGCCGCCAAGGAGGCCGCTGCTAAAGAAGCAGCAGCCA AGGAAGCAGCAGCAAAGGCCGGCTCTGCTGGTTCTGCTGCCGGGTCCGGCGAATTTGGTTCTT CTTATTACCACCATCATCATCACCACTTGGAATCCACAAGTTTATATAAGAAAGCCGGTTCTGGTT CACAAAAGGTGGAATCATTAAAACAAAAGATTGAAGAATTGAAGCAAAGAAAAGCACAATTGAAA AATGATATTGCCAATTTGGAAAAGGAAATCGCTTACGCAGAAACAGGTAGTGCCGGTTCAGCCG CGGGCTCTGGTGAATTCGCAGAAGCTGCCGCAAAAGAAGCTGCAGCAAAAGCCGGTTCTGCAG GCTCTGCTGCTGGCTCTGGCGAATTTTCCTACTATCATCATCATCATCATCACTTGGAATCCACA

AGTTTATACAAGAAAGCGGGTAGTGAATTTTTCAGAAGAGAAAGAAACAAGATGGCAGCCGCTAA GTGTAGAAACAGAAGAAGAGAATTGACTGATACATTACAAGCTGAAACAGATCAATTAGAAGACG AAAAATCAGCTTTGCAAACCGAAATCGCAAATTTGTTGAAAGAAAAAGAAAAATTGGAATTCATTT TAGCAGCCCATAGACCAGCTTGCAAAATACCTGATGACTTGGGTTTTCCAGAAGAAATGTCTTTA GAAGGTAGTGCCGGTAGTGCCGCTGGCTCAGGTGAATTTGGTAGTGCAGAAGCTGCCGCGAAA GAAGCCGCAGCTAAAGAAGCTGCCGCCAAAGAGGCAGCCGCAAAGGCAGGTTCAGCAGGTTCA GCTGCCGGGTCCGGGGAATTTGGTTCATTCCAAATGCCAGCTGACACTCCTCCACCTGCATATT TGCCACCTGAAGATCCTATGACAGGTAGTGCCGGTTCTGCTGCCGGGTCTGGCGAATTCGCTGA AGCCGCTGCTAAGGAGGCTGCAGCTAAGGCCGGCTCTGCAGGTTCCGCTGCAGGTTCAGGTGA ATTTGAAAGAGAATCTAACGAAGAACCACCTCCACCTTATGAAGATCCATACTGGGGTAATGGTG GTTCTGCCGGTAGTGCCGCCGGCTCAGGCGAATTTGGTTCTGCGGAGGCTGCTGCAAAGGAAG CTGCGGCCAAGGAAGCTGCCGCAAAAGAGGCTGCTGCCAAGGCCGGTTCAGCAGGTTCAGCA GCTGGGTCCGGTGAATTTGGTTCCAGTTATTATCACCACCATCATCACCACTTGGAATCTACCTC ATTATATAAGAAAGCGGGTTCCGGTAGTCAAAAAGTTGCAGAATTGAAAAACAGAGTTGCTGTCA AATTAAATAGAAATGAGCAGTTGAAAAATAAGGTCGAGGAGTTGAAAAATAGAAACGCATACTTG AAAAATGAATTGGCTACTTTGGAAAACGAAGTCGCAAGATTAGAAAATGATGTAGCTGAAGGCTC TGCTGGTTCCGCAGCGGGCTCAGGTGAATTCGCCGAAGCAGCCGCAAAGGAAGCTGCCGCTAA GGCCGGCTCAGCAGGTTCTGCCGCCGGAAGCGGTGAATTTTCTTATTACCACCACCACCATCAC CACTTGGAATCTACTTCATTATACAAGAAAGCGGGGTCCGGTAGTAACGAAGTCACAACCTTAGA AAATGATGCAGCCTTTATAGAAAACGAAAATGCCTACTTAGAAAAAGAAATTGCAAGATTGAGAA AGGAAAAAGCTGCATTGAGAAACAGATTAGCCCACAAGAAATCTTACTATCACCACCATCATCAT CACTTGGAATCTACATCATTATACAAGAAAGCGGGCTCCGGTAGTGCTAGAAATGCCTACTTAAG AAAGAAAATAGCCAGATTGAAGAAAGACAATTTGCAATTAGAGAGAGATGAACAGAACTTAGAAA AGATTATAGCCAATTTGAGAGATGAAATTGCTAGATTAGAAAATGAAGTAGCTTCTCATGAACAAG GTAGTGCTGGCTCCGCTGCCGGCTCCGGAGAATTTGCCGAAGCTGCCGCCAAGGAAGCCGCG GCCAAGGCTGGTTCCGCTGGTTCTGCTGCCGGATCTGGAGAATTTTCCTATTACCATCATCATCA TCATCATTTGGAATCTACATCATTATACAAGAAAGCGGGATCTGGTTCTAACTTGGTCGCCCAATT GGAGAACGAAGTCGCATCATTGGAGAACGAAAACGAAACCTTGAAGAAAAAGAACTTACACAAA AAGGATTTGATAGCTTACTTAGAAAAAGAAATCGCTAATTTGAGAAAGAAAATTGAAGAAGGTAGT GCAGGTTCAGCCGCTGGCTCCGGTGAATTTGGTTCAGCGGAGGCTGCCGCTAAGGAGGCAGCC GCTAAAGAAGCAGCCGCTAAGGAGGCTGCAGCAAAAGCAGGTTCCGCAGGTTCTGCAGCGGGT TCCGGAGAATTTGGTTCTGAACAAAAGTTGATCTCTGAAGAAGATTTGGAACAAAAGTTGATATC TGAAGAAGACTTGGAACAAAAATTAATATCAGAAGAAGATTTGGGTAGTGCAGGTTCAGCAGCTG GTTCTGGAGAATTTGGTTCAGCAGGTTCTGCCGCTGGAAGTGGCGAATTCGGTAGTGCCGGCTC CGCTGCTGGCTCTGGCGAATTTGGTTCTGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCAGGTG ACGTTGAAGAAAATCCAGGTCCAATGGGTTCTGCTGGTTCAGCTGCAGGTTCTGGTGAATTTGG TTCCGCAGGTAGTGCCGCTGGTTCTGGTGAATTCGGTTCTGCTGGTTCAGCAGCCGGTTCTGGT GAATTTTCATATTACCATCACCATCACCATCACTTGGAATCCACCAGTTTATACAAAAAGGCTGGT TCTGGTTCAGCTAGAAACGCATATTTGAGAAAGAAAATTGCTAGATTGAAGAAAGATAACTTGCA ATTGGAAAGAGATGAACAAAATTTGGAAAAGATTATCGCCAACTTAAGAGATGAAATAGCAAGAT TGGAAAACGAAGTAGCTTCTCATGAACAAGGTTCCGCAGGTAGTGCAGCTGGTTCTGGTGAATT TGCTGAAGCCGCTGCAAAGGAAGCCGCTGCAAAAGCTGGTTCCGCTGGTTCAGCCGCTGGTTC CGGTGAATTCAGTTACTATCACCATCACCATCATCACTTGGAATCCACAAGTTTATATAAAAAGGC CGGTTCTGGTTCAAATTTGGTTGCTCAATTAGAAAACGAAGTCGCATCTTTAGAAAACGAAAACG AAACATTGAAAAAGAAAAATTTGCATAAGAAAGATTTGATCGCTTATTTGGAAAAGGAAATCGCAA ACTTGAGAAAGAAAATAGAAGAAGGTTCCGCTGGTTCTGCTGCTGGTTCCGGTGAATTTGGTTCA GCTGAAGCTGCAGCCAAGGAAGCTGCAGCCAAAGAAGCCGCTGCTAAAGAAGCTGCAGCCAAA GCAGGTTCTGCCGGTTCTGCCGCAGGTTCCGGTGAATTCGGTTCTTCAGCTACTAGAGAATTGG ATGAATTGATGGCATCCTTAAGTGACTTCAAGATACAAGGTGGTTCCGCTGGTTCTGCAGCCGG CTCTGGCGAATTCGCAGAAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCTGGTTCTGCAGGTTC TGCTGCCGGTTCTGGTGAATTCGATTTGGCTTTGTCTGAAAACTGGGCACAAGAATTCTTGGCTG CAGGTGACGCTGTTGATGGTTCTGCTGGTAGTGCTGCCGGTTCAGGTGAATTTGGTAGTGCTGA

AGCTGCTGCCAAAGAAGCAGCCGCTAAAGAAGCTGCTGCCAAGGAAGCTGCCGCTAAAGCAGG TTCCGCCGGTTCTGCCGCCGGCTCCGGCGAATTTGGTTCAGATTATAAGGATGACGATGACAAG GATTACAAAGACGATGATGACAAGGATTATAAAGATGACGATGACAAAGGTTCCGCTGGTAGTG CCGCCGGCTCTGGAGAATTCGGTTCTGCCGGTTCAGCTGCCGGCTCCGGAGAATTTGGTTCCG CTGGTAGTGCAGCCGGTTCAGGTGAATTCGGTTCTGGTGCTACTAACTTCTCTTTGTTGAAGCAA GCAGGTGACGTTGAAGAAAATCCAGGTCCAATGAGTGCTAAGGCAATTTCTGAACAAACTGGTA AAGAATTGTTGTACAAGTTTATTTGTACTACATCAGCCATCCAAAATAGATTCAAATACGCTAGAG TTACCCCAGATACTGACTGGGCTAGATTGTTACAAGATCATCCATGGTTGTTATCTCAAAACTTG GTTGTCAAACCTGACCAATTAATTAAGAGAAGAGGTAAATTGGGTTTAGTAGGTGTTAATTTGACA TTGGATGGTGTAAAGTCTTGGTTGAAACCAAGATTAGGTCAAGAAGCCACAGTTGGTAAAGCTAC CGGTTTCTTGAAAAATTTCTTGATCGAACCATTTGTCCCTCATTCACAAGCCGAAGAATTCTATGT ATGTATCTACGCTACTAGAGAGGGTGACTATGTTTTATTTCATCACGAAGGTGGTGTCGACGTAG GTGACGTTGACGCCAAGGCTCAAAAGTTGTTGGTTGGTGTCGATGAAAAGTTGAACCCAGAAGA CATTAAAAAGCATTTGTTGGTTCACGCACCTGAAGATAAAAAGGAAATATTGGCCTCCTTTATAAG TGGTTTGTTTAATTTCTACGAAGATTTGTACTTCACCTACTTGGAAATTAACCCATTAGTAGTTACT AAGGATGGTGTATATGTTTTGGACTTAGCTGCAAAAGTTGATGCAACAGCCGACTACATTTGTAA GGTCAAATGGGGTGACATCGAATTTCCACCTCCATTCGGTAGAGAAGCTTATCCAGAAGAAGCC TACATTGCTGATTTGGACGCTAAGTCTGGTGCATCATTGAAGTTGACATTGTTGAACCCTAAAGG TAGAATTTGGACCATGGTTGCTGGTGGTGGTGCTAGTGTCGTATATTCTGATACTATATGCGACT TGGGTGGTGTTAACGAATTGGCAAACTACGGTGAATACTCAGGTGCCCCATCCGAACAACAAAC ATACGATTACGCTAAGACCATCTTGTCCTTAATGACTAGAGAAAAGCATCCTGATGGTAAAATCTT GATCATCGGTGGTAGTATCGCAAATTTTACTAACGTTGCCGCTACATTCAAGGGTATCGTCAGAG CTATAAGAGATTACCAAGGTCCATTGAAGGAACACGAAGTAACAATATTCGTTAGAAGAGGTGGT CCTAACTACCAAGAAGGTTTGAGAGTCATGGGTGAAGTAGGTAAAACCACTGGTATACCAATCCA TGTCTTTGGTACAGAAACCCACATGACTGCAATAGTTGGTATGGCCTTAGGTCATAGACCAATCC CTAATCAACCTCCAACCGCAGCCCACACTGCAAATTTCTTGTTAAACGCCTCTGGTTCAACTTCC ACACCAGCTCCTTCTAGAACAGCAAGTTTCTCTGAATCAAGAGCTGATGAAGTCGCTCCAGCTAA GAAAGCAAAACCAGCCATGCCTCAAGACTCCGTTCCAAGTCCTAGATCTTTGCAGGGTAAATCTA CTACTTTGTTTTCTAGACATACTAAGGCTATAGTATGGGGTATGCAAACAAGAGCAGTTCAAGGC ATGTTGGATTTCGACTATGTTTGTAGTAGAGATGAACCATCTGTTGCTGCAATGGTCTATCCTTTT ACTGGTGACCATAAGCAAAAATTCTACTGGGGTCACAAGGAAATATTGATCCCAGTTTTTAAGAA CATGGCCGATGCTATGAGAAAACATCCTGAAGTCGACGTATTGATTAACTTCGCCTCATTAAGAT CCGCTTACGATTCTACAATGGAAACCATGAACTACGCTCAAATAAGAACCATCGCTATCATTGCA GAAGGTATTCCAGAAGCCTTGACTAGAAAGTTGATTAAGAAAGCTGATCAAAAAGGTGTCACAAT AATCGGTCCAGCTACCGTAGGTGGTATTAAGCCTGGTTGTTTCAAGATCGGTAACACTGGTGGT ATGTTGGATAACATATTGGCATCTAAGTTGTATAGACCAGGTTCAGTCGCTTACGTATCCAGAAG TGGTGGTATGTCCAACGAATTGAACAACATCATCAGTAGAACTACAGATGGTGTATACGAAGGTG TTGCTATTGGTGGTGACAGATACCCAGGTTCTACTTTTATGGATCATGTATTGAGATATCAAGACA CACCTGGTGTTAAAATGATTGTTGTCTTGGGTGAAATAGGTGGTACTGAAGAATACAAGATATGC AGAGGTATCAAAGAAGGTAGATTGACAAAGCCAATCGTTTGTTGGTGCATTGGTACTTGTGCAAC AATGTTTTCTTCAGAAGTTCAATTCGGTCATGCAGGTGCCTGCGCTAATCAAGCTTCAGAAACAG CAGTTGCCAAGAACCAAGCATTAAAAGAAGCCGGTGTTTTTGTCCCTAGATCTTTCGATGAATTA GGTGAAATCATTCAATCAGTCTATGAAGACTTGGTAGCTAATGGTGTAATTGTTCCAGCACAAGA AGTTCCTCCACCTACTGTCCCTATGGATTACTCTTGGGCAAGAGAATTGGGTTTAATTAGAAAGC CAGCTAGTTTTATGACCTCTATATGTGATGAAAGAGGTCAAGAATTGATCTATGCTGGTATGCCT ATTACTGAAGTATTCAAAGAAGAAATGGGTATCGGTGGTGTTTTAGGTTTGTTGTGGTTCCAAAA GAGATTGCCAAAGTACTCTTGTCAATTCATTGAAATGTGCTTAATGGTTACAGCTGATCATGGTCC TGCTGTCTCAGGTGCACACAATACCATAATCTGCGCTAGAGCTGGTAAAGATTTGGTTTCTTCTT TGACCTCAGGTTTGTTAACTATTGGTGACAGATTTGGTGGTGCATTAGACGCCGCTGCAAAGATG TTTTCAAAAGCTTTCGATTCCGGTATAATCCCAATGGAATTCGTTAATAAGATGAAAAAGGAGGGT AAATTGATAATGGGTATCGGTCATCGTGTTAAGTCTATCAATAACCCTGATATGAGAGTACAAATC TTGAAGGACTATGTTAGACAACACTTTCCAGCCACACCTTTGTTAGATTACGCTTTGGAAGTTGA

AAAGATTACCACTTCTAAAAAGCCAAATTTGATCTTGAACGTTGATGGTTTAATTGGTGTTGCTTT TGTCGACATGTTGAGAAACTGTGGTTCCTTCACTAGAGAAGAAGCTGATGAATATATCGACATTG GTGCATTGAATGGTATCTTTGTTTTAGGTAGATCTATGGGTTTCATTGGTCATTACTTGGATCAAA AGAGATTAAAGCAAGGTTTGTACAGACATCCATGGGATGACATTTCTTACGTTTTACCTGAACAC ATGTCAATGAAATTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTG CCGAAGCTTGGTACAATTTGGGTAACGCATACTACAAGCAGGGTGACTACCAAAAGGCAATTGA ATATTACCAAAAGGCCTTGGAATTAGACCCAAATAACGCAGAAGCCTGGTATAATTTGGGTAATG CTTATTATAAACAGGGTGACTATCAAAAGGCTATCGAATACTACCAAAAGGCATTGGAATTAGAC CCTAATAACGCTGAAGCATGGTATAATTTGGGTAACGCTTATTATAAGCAGGGTGACTATCAAAA AGCCATCGAAGACTACCAAAAGGCTTTGGAATTAGATCCAAATAACTTACAAGCCGAAGCTTGGA AGAATTTGGGTAACGCTTACTATAAACAGGGTGACTACCAAAAAGCAATTGAATACTATCAAAAA GCTTTAGAATTGGACCCTAATAACGCATCAGCCTGGTACAATTTGGGTAATGCTTACTATAAGCA GGGTGACTATCAGAAGGCCATTGAATACTATCAAAAGGCTTTAGAATTGGATCCAAATAACGCTA AAGCATGGTACAGACGTGGTAACGCTTATTACAAACAGGGTGACTACCAGAAAGCCATTGAAGA TTATCAAAAGGCTTTGGAATTGGATCCTAACAACAGATCTAGATCAGCTGGTGGTGGTGGTTCTG GTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCATATTACCATCACCATCACCATCACTTGGAA TCCACAAGTTTATACAAAAAGGCTGGTTCTGGTTCAAATTTGGTCGCACAATTGGAAAACGAAGT AGCCTCTTTAGAAAATGAAAACGAAACCTTGAAAAAGAAAAACTTACATAAGAAAGATTTGATCGC TTATTTGGAAAAGGAAATCGCAAATTTGAGAAAGAAAATTGAAGAAGGTAGTGCAGGTTCTGCCG CTGGTTCTGGTGAATTTGGTTCAGCTGAAGCAGCCGCTAAGGAAGCAGCCGCTAAAGCCGGTTC AGCTGGTTCCGCAGCCGGTTCTGGTGAATTCGGTTCCAGTTACTATCACCATCACCATCATCACT TGGAATCCACAAGTTTATATAAGAAAGCAGGTTCTGGTTCAGCAAGAAATGCCTACTTGAGAAAG AAAATAGCTAGATTAAAGAAAGATAACTTGCAATTGGAAAGAGATGAACAAAATTTGGAAAAGATT ATCGCCAACTTAAGAGATGAAATCGCTAGATTGGAAAATGAAGTTGCATCCCATGAACAAGGTAG TGGTGCTACTAACTTCTCTTTGTTGAAGCAAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGA AAAACTGTGTAATCGTTTCTGCTGTTAGAACTGCAATTGGTTCCTTTAATGGTAGTTTGGCCTCTA CATCAGCTATTGATTTGGGTGCTACCGTCATCAAAGCTGCAATTGAAAGAGCAAAGATTGATTCT CAACATGTCGACGAAGTAATAATGGGTAACGTTTTGCAAGCTGGTTTAGGTCAAAATCCAGCAAG ACAAGCCTTGTTAAAATCTGGTTTAGCAGAAACTGTATGTGGTTTCACAGTTAATAAGGTCTGCG GTTCTGGTTTGAAGTCAGTTGCTTTAGCCGCTCAAGCTATACAAGCAGGTCAAGCCCAATCTATC GTCGCTGGTGGTATGGAAAATATGTCATTGGCACCTTATTTGTTAGATGCAAAAGCCAGATCAGG TTATAGATTAGGTGACGGTCAAGTATACGACGTTATTTTGAGAGATGGTTTAATGTGCGCTACTC ATGGTTATCACATGGGTATTACAGCAGAAAATGTTGCCAAAGAATACGGTATAACCAGAGAAATG CAAGATGAATTGGCATTACATTCCCAAAGAAAGGCAGCCGCTGCAATCGAAAGTGGTGCTTTTAC TGCAGAAATTGTCCCAGTAAACGTTGTCACAAGAAAGAAAACTTTCGTTTTCTCCCAAGATGAATT CCCAAAAGCTAATAGTACCGCTGAAGCATTGGGTGCTTTAAGACCTGCATTCGACAAGGCCGGT ACCGTAACTGCCGGTAATGCTTCTGGTATAAACGATGGTGCCGCTGCATTGGTTATCATGGAAG AATCAGCCGCTTTAGCAGCCGGTTTGACACCTTTAGCTAGAATTAAATCTTATGCATCAGGTGGT GTTCCACCTGCTTTGATGGGTATGGGTCCAGTCCCTGCTACCCAAAAGGCATTGCAATTAGCCG GTTTGCAATTGGCTGATATCGACTTAATCGAAGCAAACGAAGCCTTTGCTGCACAATTCTTGGCA GTTGGTAAAAATTTGGGTTTCGACTCCGAAAAGGTTAATGTCAACGGTGGTGCCATTGCTTTGGG TCATCCAATAGGTGCTTCAGGTGCAAGAATCTTGGTTACATTGTTGCATGCCATGCAAGCTAGAG ATAAAACCTTGGGTTTAGCTACTTTGTGTATCGGTGGTGGTCAAGGTATCGCAATGGTTATCGAA AGATTGAATAAGTTGTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTG CAGAAGCCTGGTACAATTTGGGTAACGCTTACTACAAGCAGGGTGACTACCAAAAGGCAATCGA ATACTACCAAAAGGCCTTGGAATTAGATCCAAATAACGCTGAAGCATGGTATAATTTGGGTAATG CCTATTATAAACAGGGTGACTATCAAAAAGCTATTGAATATTACCAAAAGGCATTGGAATTAGATC CTAATAACGCCGAAGCTTGGTATAATTTGGGTAACGCCTATTATAAGCAGGGTGACTATCAAAAG GCCATCGAAGATTACCAAAAGGCTTTGGAATTGGATCCAAACAACTTGCAAGCAGAAGCCTGGA AGAATTTGGGTAACGCTTATTACAAACAGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAA GCCTTAGAATTGGATCCTAATAACGCTTCTGCATGGTACAATTTGGGTAATGCCTACTATAAACA GGGTGACTACCAGAAGGCTATTGAATACTACCAAAAAGCATTAGAATTGGATCCAAATAACGCCA

AGGCTTGGTACAGACGTGGTAATGCCTATTACAAGCAGGGTGACTACCAGAAAGCCATAGAAGA CTATCAAAAAGCCTTGGAATTGGATCCTAACAACAGATCCAGAAGTGCTGGTGGTGGTGGTTCT GGTGGTGGTGGTTCTGGTGGTGGTGGTGCTTCTTCATATTACCATCACCATCACCATCACTTGGA ATCTACATCATTATACAAAAAGGCTGGTTCCGGTAGTAATGAAGTTACTACATTGGAAAACGATG CCGCTTTTATCGAAAACGAAAACGCATACTTGGAAAAGGAAATCGCCAGATTAAGAAAGGAAAAG GCAGCCTTGAGAAATAGATTAGCCCATAAAAAGGGTTCCGCTGGTAGTGCTGCAGGTTCTGGTG AATTTGGTTCAGCTGAAGCCGCTGCAAAAGAAGCCGCTGCAAAGGCAGGTTCTGCCGGTTCAGC CGCTGGTTCTGGTGAATTCGGTTCCAGTTACTATCACCATCACCATCATCACTTGGAATCTACTT CATTATATAAAAAGGCCGGTTCCGGTAGTCAAAAAGTCGCTGAATTAAAGAACAGAGTAGCTGTT AAGTTGAACAGAAACGAACAATTGAAAAATAAGGTAGAAGAATTGAAAAATAGAAACGCCTACTT AAAGAATGAATTGGCAACATTGGAAAACGAAGTCGCTAGATTGGAAAATGATGTAGCAGAAGGTT CTGGT

Figure 15E

Complete SOL Gene Cassette Nucleotide Sequence

ATGAGTGCTAAGGCAATTTCTGAACAAACTGGTAAAGAATTGTTGTACAAGTTTATTTGTACTACA TCAGCCATCCAAAATAGATTCAAATACGCTAGAGTTACCCCAGATACTGACTGGGCTAGATTGTT ACAAGATCATCCATGGTTGTTATCTCAAAACTTGGTTGTCAAACCTGACCAATTAATTAAGAGAAG AGGTAAATTGGGTTTAGTAGGTGTTAATTTGACATTGGATGGTGTAAAGTCTTGGTTGAAACCAA GATTAGGTCAAGAAGCCACAGTTGGTAAAGCTACCGGTTTCTTGAAAAATTTCTTGATCGAACCA TTTGTCCCTCATTCACAAGCCGAAGAATTCTATGTATGTATCTACGCTACTAGAGAGGGTGACTA TGTTTTATTTCATCACGAAGGTGGTGTCGACGTAGGTGACGTTGACGCCAAGGCTCAAAAGTTGT TGGTTGGTGTCGATGAAAAGTTGAACCCAGAAGACATTAAAAAGCATTTGTTGGTTCACGCACCT GAAGATAAAAAGGAAATATTGGCCTCCTTTATAAGTGGTTTGTTTAATTTCTACGAAGATTTGTAC TTCACCTACTTGGAAATTAACCCATTAGTAGTTACTAAGGATGGTGTATATGTTTTGGACTTAGCT GCAAAAGTTGATGCAACAGCCGACTACATTTGTAAGGTCAAATGGGGTGACATCGAATTTCCACC TCCATTCGGTAGAGAAGCTTATCCAGAAGAAGCCTACATTGCTGATTTGGACGCTAAGTCTGGTG CATCATTGAAGTTGACATTGTTGAACCCTAAAGGTAGAATTTGGACCATGGTTGCTGGTGGTGGT GCTAGTGTCGTATATTCTGATACTATATGCGACTTGGGTGGTGTTAACGAATTGGCAAACTACGG TGAATACTCAGGTGCCCCATCCGAACAACAAACATACGATTACGCTAAGACCATCTTGTCCTTAA TGACTAGAGAAAAGCATCCTGATGGTAAAATCTTGATCATCGGTGGTAGTATCGCAAATTTTACT AACGTTGCCGCTACATTCAAGGGTATCGTCAGAGCTATAAGAGATTACCAAGGTCCATTGAAGG AACACGAAGTAACAATATTCGTTAGAAGAGGTGGTCCTAACTACCAAGAAGGTTTGAGAGTCATG GGTGAAGTAGGTAAAACCACTGGTATACCAATCCATGTCTTTGGTACAGAAACCCACATGACTGC AATAGTTGGTATGGCCTTAGGTCATAGACCAATCCCTAATCAACCTCCAACCGCAGCCCACACTG CAAATTTCTTGTTAAACGCCTCTGGTTCAACTTCCACACCAGCTCCTTCTAGAACAGCAAGTTTCT CTGAATCAAGAGCTGATGAAGTCGCTCCAGCTAAGAAAGCAAAACCAGCCATGCCTCAAGACTC CGTTCCAAGTCCTAGATCTTTGCAGGGTAAATCTACTACTTTGTTTTCTAGACATACTAAGGCTAT AGTATGGGGTATGCAAACAAGAGCAGTTCAAGGCATGTTGGATTTCGACTATGTTTGTAGTAGAG ATGAACCATCTGTTGCTGCAATGGTCTATCCTTTTACTGGTGACCATAAGCAAAAATTCTACTGG GGTCACAAGGAAATATTGATCCCAGTTTTTAAGAACATGGCCGATGCTATGAGAAAACATCCTGA AGTCGACGTATTGATTAACTTCGCCTCATTAAGATCCGCTTACGATTCTACAATGGAAACCATGA ACTACGCTCAAATAAGAACCATCGCTATCATTGCAGAAGGTATTCCAGAAGCCTTGACTAGAAAG TTGATTAAGAAAGCTGATCAAAAAGGTGTCACAATAATCGGTCCAGCTACCGTAGGTGGTATTAA GCCTGGTTGTTTCAAGATCGGTAACACTGGTGGTATGTTGGATAACATATTGGCATCTAAGTTGT ATAGACCAGGTTCAGTCGCTTACGTATCCAGAAGTGGTGGTATGTCCAACGAATTGAACAACATC ATCAGTAGAACTACAGATGGTGTATACGAAGGTGTTGCTATTGGTGGTGACAGATACCCAGGTT CTACTTTTATGGATCATGTATTGAGATATCAAGACACACCTGGTGTTAAAATGATTGTTGTCTTGG GTGAAATAGGTGGTACTGAAGAATACAAGATATGCAGAGGTATCAAAGAAGGTAGATTGACAAA GCCAATCGTTTGTTGGTGCATTGGTACTTGTGCAACAATGTTTTCTTCAGAAGTTCAATTCGGTCA TGCAGGTGCCTGCGCTAATCAAGCTTCAGAAACAGCAGTTGCCAAGAACCAAGCATTAAAAGAA GCCGGTGTTTTTGTCCCTAGATCTTTCGATGAATTAGGTGAAATCATTCAATCAGTCTATGAAGAC TTGGTAGCTAATGGTGTAATTGTTCCAGCACAAGAAGTTCCTCCACCTACTGTCCCTATGGATTA CTCTTGGGCAAGAGAATTGGGTTTAATTAGAAAGCCAGCTAGTTTTATGACCTCTATATGTGATG AAAGAGGTCAAGAATTGATCTATGCTGGTATGCCTATTACTGAAGTATTCAAAGAAGAAATGGGT ATCGGTGGTGTTTTAGGTTTGTTGTGGTTCCAAAAGAGATTGCCAAAGTACTCTTGTCAATTCATT GAAATGTGCTTAATGGTTACAGCTGATCATGGTCCTGCTGTCTCAGGTGCACACAATACCATAAT CTGCGCTAGAGCTGGTAAAGATTTGGTTTCTTCTTTGACCTCAGGTTTGTTAACTATTGGTGACA GATTTGGTGGTGCATTAGACGCCGCTGCAAAGATGTTTTCAAAAGCTTTCGATTCCGGTATAATC CCAATGGAATTCGTTAATAAGATGAAAAAGGAGGGTAAATTGATAATGGGTATCGGTCATCGTGT TAAGTCTATCAATAACCCTGATATGAGAGTACAAATCTTGAAGGACTATGTTAGACAACACTTTCC AGCCACACCTTTGTTAGATTACGCTTTGGAAGTTGAAAAGATTACCACTTCTAAAAAGCCAAATTT GATCTTGAACGTTGATGGTTTAATTGGTGTTGCTTTTGTCGACATGTTGAGAAACTGTGGTTCCTT CACTAGAGAAGAAGCTGATGAATATATCGACATTGGTGCATTGAATGGTATCTTTGTTTTAGGTA GATCTATGGGTTTCATTGGTCATTACTTGGATCAAAAGAGATTAAAGCAAGGTTTGTACAGACAT

CCATGGGATGACATTTCTTACGTTTTACCTGAACACATGTCAATGAAATTGTCTGGTGGTGGTGG TTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCCGAAGCTTGGTACAATTTGGGTAACGCA TACTACAAGCAGGGTGACTACCAAAAGGCAATTGAATATTACCAAAAGGCCTTGGAATTAGACCC AAATAACGCAGAAGCCTGGTATAATTTGGGTAATGCTTATTATAAACAGGGTGACTATCAAAAGG CTATCGAATACTACCAAAAGGCATTGGAATTAGACCCTAATAACGCTGAAGCATGGTATAATTTG GGTAACGCTTATTATAAGCAGGGTGACTATCAAAAAGCCATCGAAGACTACCAAAAGGCTTTGGA ATTAGATCCAAATAACTTACAAGCCGAAGCTTGGAAGAATTTGGGTAACGCTTACTATAAACAGG GTGACTACCAAAAAGCAATTGAATACTATCAAAAAGCTTTAGAATTGGACCCTAATAACGCATCA GCCTGGTACAATTTGGGTAATGCTTACTATAAGCAGGGTGACTATCAGAAGGCCATTGAATACTA TCAAAAGGCTTTAGAATTGGATCCAAATAACGCTAAAGCATGGTACAGACGTGGTAACGCTTATT ACAAACAGGGTGACTACCAGAAAGCCATTGAAGATTATCAAAAGGCTTTGGAATTGGATCCTAAC AACAGATCTAGATCAGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTGCTT CTTCATATTACCATCACCATCACCATCACTTGGAATCCACAAGTTTATACAAAAAGGCTGGTTCTG GTTCAAATTTGGTCGCACAATTGGAAAACGAAGTAGCCTCTTTAGAAAATGAAAACGAAACCTTG AAAAAGAAAAACTTACATAAGAAAGATTTGATCGCTTATTTGGAAAAGGAAATCGCAAATTTGAGA AAGAAAATTGAAGAAGGTAGTGCAGGTTCTGCCGCTGGTTCTGGTGAATTTGGTTCAGCTGAAG CAGCCGCTAAGGAAGCAGCCGCTAAAGCCGGTTCAGCTGGTTCCGCAGCCGGTTCTGGTGAAT TCGGTTCCAGTTACTATCACCATCACCATCATCACTTGGAATCCACAAGTTTATATAAGAAAGCAG GTTCTGGTTCAGCAAGAAATGCCTACTTGAGAAAGAAAATAGCTAGATTAAAGAAAGATAACTTG CAATTGGAAAGAGATGAACAAAATTTGGAAAAGATTATCGCCAACTTAAGAGATGAAATCGCTAG ATTGGAAAATGAAGTTGCATCCCATGAACAAGGTAGTGGTGCTACTAACTTCTCTTTGTTGAAGC AAGCAGGTGACGTTGAAGAAAATCCAGGTCCAATGAAAAACTGTGTAATCGTTTCTGCTGTTAGA ACTGCAATTGGTTCCTTTAATGGTAGTTTGGCCTCTACATCAGCTATTGATTTGGGTGCTACCGT CATCAAAGCTGCAATTGAAAGAGCAAAGATTGATTCTCAACATGTCGACGAAGTAATAATGGGTA ACGTTTTGCAAGCTGGTTTAGGTCAAAATCCAGCAAGACAAGCCTTGTTAAAATCTGGTTTAGCA GAAACTGTATGTGGTTTCACAGTTAATAAGGTCTGCGGTTCTGGTTTGAAGTCAGTTGCTTTAGC CGCTCAAGCTATACAAGCAGGTCAAGCCCAATCTATCGTCGCTGGTGGTATGGAAAATATGTCAT TGGCACCTTATTTGTTAGATGCAAAAGCCAGATCAGGTTATAGATTAGGTGACGGTCAAGTATAC GACGTTATTTTGAGAGATGGTTTAATGTGCGCTACTCATGGTTATCACATGGGTATTACAGCAGA AAATGTTGCCAAAGAATACGGTATAACCAGAGAAATGCAAGATGAATTGGCATTACATTCCCAAA GAAAGGCAGCCGCTGCAATCGAAAGTGGTGCTTTTACTGCAGAAATTGTCCCAGTAAACGTTGT CACAAGAAAGAAAACTTTCGTTTTCTCCCAAGATGAATTCCCAAAAGCTAATAGTACCGCTGAAG CATTGGGTGCTTTAAGACCTGCATTCGACAAGGCCGGTACCGTAACTGCCGGTAATGCTTCTGG TATAAACGATGGTGCCGCTGCATTGGTTATCATGGAAGAATCAGCCGCTTTAGCAGCCGGTTTG ACACCTTTAGCTAGAATTAAATCTTATGCATCAGGTGGTGTTCCACCTGCTTTGATGGGTATGGG TCCAGTCCCTGCTACCCAAAAGGCATTGCAATTAGCCGGTTTGCAATTGGCTGATATCGACTTAA TCGAAGCAAACGAAGCCTTTGCTGCACAATTCTTGGCAGTTGGTAAAAATTTGGGTTTCGACTCC GAAAAGGTTAATGTCAACGGTGGTGCCATTGCTTTGGGTCATCCAATAGGTGCTTCAGGTGCAA GAATCTTGGTTACATTGTTGCATGCCATGCAAGCTAGAGATAAAACCTTGGGTTTAGCTACTTTGT GTATCGGTGGTGGTCAAGGTATCGCAATGGTTATCGAAAGATTGAATAAGTTGTCTGGTGGTGG TGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGCAGAAGCCTGGTACAATTTGGGTAAC GCTTACTACAAGCAGGGTGACTACCAAAAGGCAATCGAATACTACCAAAAGGCCTTGGAATTAG ATCCAAATAACGCTGAAGCATGGTATAATTTGGGTAATGCCTATTATAAACAGGGTGACTATCAA AAAGCTATTGAATATTACCAAAAGGCATTGGAATTAGATCCTAATAACGCCGAAGCTTGGTATAAT TTGGGTAACGCCTATTATAAGCAGGGTGACTATCAAAAGGCCATCGAAGATTACCAAAAGGCTTT GGAATTGGATCCAAACAACTTGCAAGCAGAAGCCTGGAAGAATTTGGGTAACGCTTATTACAAAC AGGGTGACTACCAAAAAGCTATTGAATACTATCAAAAAGCCTTAGAATTGGATCCTAATAACGCTT CTGCATGGTACAATTTGGGTAATGCCTACTATAAACAGGGTGACTACCAGAAGGCTATTGAATAC TACCAAAAAGCATTAGAATTGGATCCAAATAACGCCAAGGCTTGGTACAGACGTGGTAATGCCTA TTACAAGCAGGGTGACTACCAGAAAGCCATAGAAGACTATCAAAAAGCCTTGGAATTGGATCCTA ACAACAGATCCAGAAGTGCTGGTGGTGGTGGTTCTGGTGGTGGTGGTTCTGGTGGTGGTGGTG CTTCTTCATATTACCATCACCATCACCATCACTTGGAATCTACATCATTATACAAAAAGGCTGGTT

CCGGTAGTAATGAAGTTACTACATTGGAAAACGATGCCGCTTTTATCGAAAACGAAAACGCATAC TTGGAAAAGGAAATCGCCAGATTAAGAAAGGAAAAGGCAGCCTTGAGAAATAGATTAGCCCATA AAAAGGGTTCCGCTGGTAGTGCTGCAGGTTCTGGTGAATTTGGTTCAGCTGAAGCCGCTGCAAA AGAAGCCGCTGCAAAGGCAGGTTCTGCCGGTTCAGCCGCTGGTTCTGGTGAATTCGGTTCCAGT TACTATCACCATCACCATCATCACTTGGAATCTACTTCATTATATAAAAAGGCCGGTTCCGGTAGT CAAAAAGTCGCTGAATTAAAGAACAGAGTAGCTGTTAAGTTGAACAGAAACGAACAATTGAAAAA TAAGGTAGAAGAATTGAAAAATAGAAACGCCTACTTAAAGAATGAATTGGCAACATTGGAAAACG AAGTCGCTAGATTGGAAAATGATGTAGCAGAAGGTTCTGGT

<img file="IL283384A_D0303.tif" />

CEN/ARS: 3519-4022 bp

CmR: 4432-5091 bp cat promoter: 5092-5194 bp oriV: 6052-6666 bp 0ri2: 6742-6961 bp repE: 7052-7807 bp spoA: 8386-9561 bp sopB: 9561-157 bp

FIG. 16

SUBSTITUTE SHEET (RULE 26)

Sac I Bgl II

<img file="IL283384A_D0304.tif" />

CYC1 terminator

FIG. 17

FLAG

Cal I Spe I Not I EcoRI pESC-TRP 6.5 kb

BamH I Apa I Sma I Sal I c-Myc

Xhol Kpn I Sac II Nhel

SUBSTITUTE SHEET (RULE 26)

FIGURE 18

Proliferation Rates (30° C + 400 RPM)

<img file="IL283384A_D0305.tif" />

Time (h)

ODAD1 B, Sig=205,16 Ref=360,100 (1A110819\1A110819 2019 08 09 25M108190000051.D)

<img file="IL283384A_D0306.tif" />

<img file="IL283384A_D0307.tif" />

FIG. 19A

SUBSTITUTE SHEET (RULE 26)

ODAD1 B, Sig=205,16 Ref=360,100 (1A110819\1A110819 2019 08 09 25\1108190000079.D)

8986&lt;

9388&lt;

aao-991/ε vaao-9063^

8L3&amp;#1470;L co

CXI

<img file="IL283384A_D0308.tif" />

FIG. 19B

SUBSTITUTE SHEET (RULE 26)

Total Cannabinoids

S 40 &amp;#1498;

**** -35 &amp;#1504;&amp;#1505;

23.61 &amp;#1470;30 £

<img file="IL283384A_D0309.tif" />

yCBsoL yCBscF Strain

FIG. 19C

<img file="IL283384A_D0310.tif" />

FIG. 19D

<img file="IL283384A_D0311.tif" />

iza coo(H) Ε&amp;#911; ACOOH

FIG. 19E

SUBSTITUTE SHEET (RULE 26)

FIGURE 20

Precursor-Buffered Media Cannabinoid Titers ****

<img file="IL283384A_D0312.tif" />

FIGURE 21

B Cannabinoid Biosynthesis Parameteters

[Total Cannabinoids] (mg/L)

A Citrate Cannabinoid Biosynthetic Kinetics

<img file="IL283384A_D0313.tif" />

[Buffered Citrate, pH δ.0] (mg/L)

<img file="IL283384A_D0314.tif" />

<img file="IL283384A_D0315.tif" />

EC5q [Citrate] (mg/L)

Parameter Estimate