NL2030206B1 - Heterologous expression of enzymes for increased b-farnesene production - Google Patents

Abstract

39 AB STRACT Provided are heterologous enzymes Which are capable of improving titers of farnesene products and its derivatives in applications such as fragrances, surfactants, stabilizers, resins, and 5 coatings, among others.

Description

HETEROLOGOUS EXPRESSION OF ENZYMES FOR INCREASED B-FARNESENE

PRODUCTION

SEQUENCE LISTING

[0001] The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCH copy, created on October 25, 2021, is named Sequence-Listing_ST25.txt and is 255,939 bytes in size.

TECHNICAL FIELD

[0002] Provided are methods of using heterologous enzymes which can be used to improve titers of farnesene products and its derivatives.

BACKGROUND

[0003] (E)-B-Farnesene is a sesquiterpene molecule that occurs in a wide range of both plant and animal taxa. In nature, [B-Farnesene is an alarm pheromone that deters aphids. The synthetic biology sector has also shown that it can be used in a wide array of applications such as fragrances, surfactants, stabilizers, resins, and coatings.

[0004] Yeast use the mevalonate-dependent (MEV) pathway to convert acetyl coenzyme

A (acetyl-CoA) to IPP (Isopentenyl pyrophosphate). IPP can be further metabolized to Geranyl pyrophosphate (GPP) and/or farnesyl pyrophosphate (FPP).

[0005] Historically, sesquiterpene synthases, including 3-Farnesene synthase, have been difficult to identify by typical homology and HMM derived searching methodologies. The level of homology between deduced amino acid sequences of synthases showing identical substrate and product specificity from different species can be lower than the homology between synthases catalyzing the formation of completely different terpene structural types from the same plant.

SUMMARY

[0006] The disclosure provides a eukaryotic host cell comprising one or more heterologous nucleic acids, wherein the heterologous nucleic acids alter the level of activity in the host cell of (a) one or more mevalonate pathway enzymes; and/or (b) a prenyl transferase; and wherein the host cell further comprises a heterologous -farnesene synthase and is capable of making a sesquiterpene from the action of the B-farnesene synthase. In one embodiment, the one or more mevalonate pathway enzymes is a 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR). In a further embodiment, the HMGR is a truncated HMGR. In yet another or further embodiment, the prenyl transferase is a farnesyl pyrophosphate synthase (FPPS). In a further embodiment, the FPPS is encoded by ERG20. In another or further embodiment, the heterologous nucleic acid comprises at least one additional copy of a polynucleotide encoding the FPPS. In another embodiment, the host cell produces at least a 10% more (E)-B-farnesene compared to a control. In a further embodiment, the control is the amount of (E)-B-farnesene produced by

Artemisia Annua. In still another embodiment, the heterologous B-farnesene synthase comprises a sequence having at least 50%, 60%, 70%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity, or greater sequence identity to SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and can convert (2E, 6E)-farnesyl diphosphate to (E)-beta farnesene + diphosphate. In one embodiment, the B-farnesene synthase comprises one or more mutations at a position selected from the group consisting of V12, E59, T67, A68, K70, H76, A80, F120, K136, A213, V270, A330, 1336, V350, P374, G469, N471, 1532 and T555 relative to SEQ ID NO: 20 (or at a corresponding position identifiable via protein alignment in a sequence of SEQ ID NO:26, 28, 34, 36, 38 or 42).

In another embodiment, the [-farnesene synthase comprises a sequence of SEQ ID NO:20 and further comprises one or more mutations selected from the group consisting of VI2E, E59K,

T67G, TOTP, A68D, AG8N, A68P, K70A, K70S, K70C, H76S, H76T, ASOM, ASOT, F120L.,

K1361 K136T, A213C, V270L, A330F, I336E, V350A, V350M, P3748, G469Q, N4711, 1532M and TSS55E. In still another embodiment, a B-farnesene synthase of the disclosure can be N- terminally truncated by 1 to about 20 amino acids (e.g., 1, 2,3, 4, 5,6, 7,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 amino acids) compared to the wild-type sequence and can comprise similar or improved [}-farnesene synthase activity. In another embodiment, the heterologous 3- farnesene synthase is encoded by a polynucleotide that has at least 50%, 60%, 70%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity or greater sequence identity to a polynucleotide sequence of SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41 and which encodes a polypeptide that convert (2E, 6E)-farnesyl diphosphate to (E)-beta farnesene + diphosphate. In still another embodiment, the heterologous p-farnesene synthase comprises a sequence selected from SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42. In another embodiment, the heterologous B-farnesene synthase is encoded by a codon optimized polynucleotide sequence having at least 70% sequence identity, preferably 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity to a polynucleotide sequence of SEQ ID NO: 19, 21, 23, 25, 27,29, 31, 33, 35, 37, 39, or 41.

[0007] The disclosure also provides a eukaryotic host cell comprising one or more heterologous nucleic acids, wherein the heterologous nucleic acids encode: (a) an enzyme that converts 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) to mevalonate; and/or (b) a prenyl transferase; and wherein the host cell further comprises a heterologous B-farnesene synthase and is capable of making a sesquiterpene from the action of the B-farnesene synthase. In one embodiment, the enzyme that converts HMG-CoA to mevalonate is a 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR). In still another embodiment, the enzyme that converts HMG-

CoA to mevalonate is a truncated HMGR. In another or further embodiment, expression of the

HMGR 1s under inducible control. In another embodiment, the HMGR is under constitutive control. In yet another embodiment, the prenyl transferase is a farnesyl pyrophosphate synthase (FPPS). In another embodiment, the prenyl transferase is encoded by the ERG20 gene. In still another embodiment, expression of the prenyl transferase is under inducible control. In another embodiment, the prenyl transferase is under constitutive control. In yet another embodiment, the heterologous B-farnesene synthase comprises a sequence having at least 70% sequence identity, preferably 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity to SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and can convert (2E. 6E)-farnesyl diphosphate to (E)-beta farnesene + diphosphate. In another embodiment, the heterologous B-farnesene synthase is encoded by a polynucleotide that has at least 70% sequence identity, preferably 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater to a polynucleotide sequence of SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41 and which encodes a polypeptide that convert (2E, 6E)-farnesyl diphosphate to (E)-beta farnesene + diphosphate. In still another embodiment, the heterologous p-farnesene synthase comprises a sequence selected from SEQ ID

NO:20, 26, 28, 34, 36, 38 or 42. In one embodiment, the B-farnesene synthase comprises one or more mutations at a position selected from the group consisting of V12, E59, T67, A68, K70, H76,

A80, F120, K136, A213, V270, A330, 1336, V350, P374, G469, N471, 1532 and T555 relative to

SEQ ID NO:20 (or at a corresponding position identifiable via protein alignment in a sequence of

SEQ ID NO:26, 28, 34, 36, 38 or 42). In another embodiment, the B-farnesene synthase comprises a sequence of SEQ ID NO:20 and further comprises one or more mutations selected from the group consisting of VI2E, E59K, T67G, T67P, A68D, A68N, A68P, K70A, K70S, K70C, H76S, H76T,

A80M, ASOT, F120L, K1361, K136T, A213C, V270L, A330F, 1336E, V350A, V350M, P3748,

G469Q, N4711, 1532M and T555E. In still another embodiment, a 3-farnesene synthase of the disclosure can be N-terminally truncated by 1 to about 20 amino acids (e.g., 1,2,3,4,.5,6,7. 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 amino acids) compared to the wild-type sequence and can comprise similar or improved B-farnesene synthase activity. In still another embodiment, the heterologous f-farnesene synthase is encoded by a codon optimized polynucleotide sequence having at least 70% sequence identity, preferably 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater to a polynucleotide sequence of SEQ ID NO: 19, 21,23, 25,27, 29, 31, 33, 35, 37, 39, or 41.

[0008] The disclosure also provides a Saccharomyces cerevisiae cell comprising one or more heterologous nucleic acids, wherein the heterologous nucleic acids encode: (a) a truncated 3- hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR); and/or (b) a prenyl transferase; and a heterologous B-farnesene synthase and is capable of making a sesquiterpene from the action of the

B-farnesene synthase.

[0009] The disclosure provides an isolated polypeptide comprising a sequence that has at least 50%, 60%, 70%, preferably 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or greater sequence identity to any one of SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and can convert (2E, 6E)-farnesyl diphosphate to (E)-beta farnesene + diphosphate. In one embodiment, the isolated polypeptide comprises one or more mutations at a position selected from the group consisting of V12, E59, T67, A68, K70, H76, A80, F120, K136, A213, V270, A330, 1336, V350,

P374, G469, N471, 1532 and T555 relative to SEQ ID NO:20 (or at a corresponding position identifiable via protein alignment in a sequence of SEQ ID NO:26, 28, 34, 36, 38 or 42). In another embodiment, the polypeptide comprises a sequence of SEQ ID NO:20 and further comprises one or more mutations selected from the group consisting of VI2E, E59K, T67G, T67P,

A68D, AGEN, AGSP, K70A, K70S, K70C, H76S, H76T, A80M, A80T, F120L, K1361, K136T,

A213C, V270L, A330F, I336E, V350A, V350M, P3748, G469Q, N4711, 1532M and TS55E. In still another embodiment, an isolated polypeptide of the disclosure can be N-terminally truncated by 1 to about 20 amino acids (e.g., 1,2, 3,4,5,6,7,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, or 22 amino acids) compared to the wild-type sequence and can comprise similar or improved p- farnesene synthase activity.

[0010] The disclosure also provides a codon optimized isolated polynucleotide encoding a polypeptide of the disclosure as well as vectors comprising polynucleotides encoding polypeptides of the disclosure.

[0011] The disclosure also provides a host cell comprising a polynucleotide or vector of the disclosure, wherein the polynucleotide is heterologous to the host cell. In one embodiment, the host cell further comprises a truncated 3-hydroxy-3-methylglutaryl coenzyme-A reductase (HMGR). In still another or further embodiment, the host cell further comprises at least two copies of ERG20 coding sequence. In still another or further embodiment, the host cell is eukaryotic. In a further embodiment, the host cell is a yeast cell.

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The accompanying drawings, which are incorporated into and constitute a part of this specification, illustrate one or more embodiments of the disclosure and, together with the detailed description, serve to explain the principles and implementations of the invention.

[0013] Figure 1 provides a general diagram of a recombinant pathway of the disclosure.

[0014] Figure 2 provides raw farnesene synthesis measurements for each of p-farnesene synthase of the disclosure.

[0015] Figure 3 provides a depiction of best-performing B-farnesene synthases of the disclosure.

[0016] Figure 4 shows enzymatic engineering hits on CjFS.

[0017] Figure 5 shows enzymatic truncation modifications and titers.

[0018] Figure 6 shows mean farnesene titer for CjFS RR1 parents and mutants,

DETAILED DESCRIPTION

[0019] As used herein and in the appended claims. the singular forms “a” “an” and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference 5 to "a polynucleotide" includes a plurality of such polynucleotides and reference to “the enzyme" includes reference to one or more enzymes, and so forth.

[0020] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices and materials are described herein.

[0021] Also, the use of “or” means “and/or” unless stated otherwise. Similarly, “comprise,” “comprises,” “comprising” “include,” “includes,” and “including” are interchangeable and not intended to be limiting.

[0022] 1t is to be further understood that where descriptions of various embodiments use the term “comprising,” those skilled in the art would understand that in some specific instances. an embodiment can be altematively described using language “consisting essentially of” or “consisting of.”

[0023] Any publications discussed above and throughout the text are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior disclosure.

[0024] As used herein, an "activity" of an enzyme is a measure of its ability to catalyze a reaction resulting in a metabolite, i.e., to "function", and may be expressed as the rate at which the metabolite of the reaction is produced. For example, enzyme activity can be represented as the amount of metabolite produced per unit of time or per unit of enzyme (e.g., concentration or weight), or in terms of affinity or dissociation constants.

[0025] The term “(E)-p-Famesne”, “Farnesene™, or “o-Farnesene” will refer to as a set of six closely related chemical compounds which all are sesquiterpenes. a-Famesene and B-farnesene are isomers, differing by the location of one double bond. a-Farnesene is 3,7, 11-trimethyl-1,3,6,10- dodecatetraene and pB-farnesene is 7,1 1-dimethyl-3-methylene-1,6,10-dodecatriene. The alpha form can exist as a four stereoisomers that differ about the geometry of two of its three internal double bonds (the stereoisomers of the third internal double bond are identical). The beta isomer exists as two stereoisomers about the geometry of its central double bond.

[0026] The term "biosynthetic pathway", also referred to as "metabolic pathway", refers to a set of anabolic or catabolic biochemical reactions for converting (transmuting) one chemical species into another. Gene products belong to the same "metabolic pathway" if they, in parallel or in series, act on the same substrate, produce the same product, or act on or produce a metabolic intermediate (i.e., metabolite) between the same substrate and metabolite end product. The disclosure provides recombinant microorganism having a metabolically engineered pathway for the production of a desired product or intermediate.

[0027] A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged or polar side chains {e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). The following six groups each contain amino acids that are conservative substitutions for one another: 1) Serine (8), Threonine (T); 2) Aspartic Acid (D), Glutamic Acid (E); 3) Asparagine (N),

Glutamine (Q); 4) Arginine (R), Lysine (K); 5) Isoleucine (1), Leucine (L), Methionine (M),

Alanine {A), Valine (V), and 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W).

[0028] An "enzyme" means any substance, typically composed wholly or largely of amino acids making up a protein or polypeptide that catalyzes or promotes, more or less specifically, one or more chemical or biochemical reactions.

[0029] The term "expression" with respect to a gene or polynucleotide refers to transcription of the gene or polynucleotide and, as appropriate, translation of the resulting mRNA transcript to a protein or polypeptide. Thus, as will be clear from the context, expression of a protein or polypeptide results from transcription and translation of the open reading frame.

[0030] An “expression control sequence” means a nucleic acid sequence that directs transcription of a nucleic acid to which it is operably linked. An expression control sequence can be a promoter, such as a constitutive or an inducible promoter, or an enhancer. An expression control sequence can be “native” or heterologous. A native expression control sequence is derived from the same organism, species, or strain as the gene being expressed. A heterologous expression control sequence is derived from a different organism, species, or strain as the gene being expressed. An "inducible promoter” is a promoter that is active under environmental or developmental regulation.

[0031] A “heterologous nucleic acid” or “heterologous polynucleotide™ is a polynucleotide derived from a different organism, species or strain than the host cell.

[0032] A "heterologous polypeptide” is meant a polypeptide encoded by a nucleic acid sequence derived from a different organism, species, or strain than that of the host cell. In some embodiments, a heterologous polypeptide is a variant or mutant of a wild-type polypeptide that is found in the host cell, wherein the variant or mutant is engineered to be expressed in the host cell.

[0033] A protein has "homology" or is "homologous" to a second protein if the nucleic acid sequence that encodes the protein has a similar sequence to the nucleic acid sequence that encodes the second protein. Alternatively, a protein has homology to a second protein if the two proteins have "similar" amino acid sequences. (Thus, the term "homologous proteins” is defined to mean that the two proteins have similar amino acid sequences).

[0034] As used herein, two proteins (or a region of the proteins) or two nucleic acids (or a region of nucleic acids) are substantially homologous when the amino acid sequences have at least about 30%, 40%, 50% 60%, 65%, 70%, 75%, 80%, 85%, 90%. 91%, 92%, 93%. 94%, 95%. 96%, 97%, 98%, or 99% identity. To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes).

In one embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, typically at least 40%, more typically at least 50%, even more typically at least 60%, and even more typically at least 70%, 80%. 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid "identity" is equivalent to amino acid or nucleic acid "homology"). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0035] When "homologous" is used in reference to proteins or peptides, it is recognized that residue positions that are not identical often differ by conservative amino acid substitutions. A "conservative amino acid substitution” is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of homology may be adjusted upwards to correct for the conservative nature of the substitution.

Means for making this adjustment are well known to those of skill in the art (see, e.g., Pearson ef al, 1994, hereby incorporated herein by reference).

[0036] In addition, and as mentioned above, homologs of enzymes useful for generating metabolites are encompassed by the microorganisms and methods provided herein. The term "homologs" used with respect to an original enzyme or gene of a first family or species refers to distinct enzymes or genes of a second family or species which are determined by functional, structural or genomic analyses to be an enzyme or gene of the second family or species which corresponds to the original enzyme or gene of the first family or species. Most often, homologs will have functional, structural or genomic similarities. Techniques are known by which homologs of an enzyme or gene can readily be cloned using genetic probes and PCR. Identity of cloned sequences as homolog can be confirmed using functional assays and/or by genomic mapping of the genes.

[0037] Sequence homology for polypeptides, which can also be referred to as percent sequence identity, is typically measured using sequence analysis software. See, e.g., the Sequence

Analysis Software Package of the Genetics Computer Group (GCG), University of Wisconsin

Biotechnology Center, 910 University Avenue, Madison, Wis. 53705. Protein analysis software matches similar sequences using measure of homology assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, GCG contains programs such as "Gap" and "Bestfit" which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild type protein and a mutein thereof. See, e.g., GCG Version 6.1,

[0038] A typical algorithm used comparing a molecule sequence to a database containing a large number of sequences from different organisms is the computer program BLAST (Altschul, 1990; Gish, 1993; Madden, 1996; Altschul, 1997; Zhang, 1997), especially blastp or tblastn (Altschul, 1997). Typical parameters for BLASTp are: Expectation value: 10 (default); Filter: seg (default); Cost to open a gap: 11 (default); Cost to extend a gap: 1 (default); Max. alignments: 100 (default); Word size: 11 (default); No. of descriptions: 100 (default); Penalty Matrix:

BLOWSUMS62.

[0039] When searching a database containing sequences from a large number of different organisms, it is typical to compare amino acid sequences. Database searching using amino acid sequences can be measured by algorithms other than BLASTp known in the art. For instance, polypeptide sequences can be compared using FASTA, a program in GCG Version 6.1. FASTA provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson, 1990, hereby incorporated herein by reference). For example, percent sequence identity between amino acid sequences can be determined using

FASTA with its default parameters (a word size of 2 and the PAM250 scoring matrix), as provided in GCG Version 6.1, hereby incorporated herein by reference.

[0040] The disclosure provides accession numbers and sequences for various genes, homologs and variants useful in the generation of recombinant microorganism and proteins for use in in vitro systems. It is to be understood that homologs and variants described herein are exemplary and non-limiting. Additional homologs, variants and sequences are available to those of skill in the art using various databases including, for example, the National Center for

Biotechnology Information (NCBI) access to which is available on the World-Wide-Web.

[0041] It is well within the level of skill in the art to utilize the sequences and accession number described herein to identify homologs and isozymes that can be used or substituted for any of the polypeptides used herein. In fact, a BLAST search of any one of the sequences provide herein will identify a plurality of related homologs.

[0042] An “isoprenoid precursor’ refers to any molecule that is used by organisms or part of a biosynthetic pathway in the biosynthesis of terpenoids or isoprenoids. Non-limiting examples of isoprenoid precursor molecules include, e.g., isopenteny! pyrophosphate (IPP) and dimethylallyl diphosphate (DMAPP).

[0043] “Isozymes™ refer to two polypeptides that carry out the same functional conversion/reaction, but which are so dissimilar in structure that they are typically determined to not be “homologous™.

[0044] As used herein, the term "metabolically engineered" or "metabolic engineering" involves rational pathway design and assembly of biosynthetic genes, genes associated with operons, and control elements of such polynucleotides, for the production of a desired metabolite, such as a terpenoid, or other chemical, in a microorganism. "Metabolically engineered” can further include optimization of metabolic flux by regulation and optimization of transcription, translation, protein stability and protein functionality using genetic engineering and appropriate culture condition including the reduction of, disruption, or knocking out of, a competing metabolic pathway that competes with an intermediate leading to a desired pathway. A biosynthetic gene can be heterologous to the host microorganism, either by virtue of being foreign to the host, or being modified by mutagenesis, recombination, and/or association with a heterologous expression control sequence in an endogenous host cell. In one embodiment, where the polynucleotide is xenogenetic to the host organism, the polynucleotide can be codon optimized.

[0045] A "metabolite" refers to any substance produced by metabolism or a substance necessary for or taking part in a particular metabolic process that gives rise to a desired metabolite, chemical, etc. A metabolite can be an organic compound that is a starting material {e.g., glucose etc.), an intermediate in {e.g., acetyl-coA), or an end product (e.g., sesquiterpene) of metabolism.

Metabolites can be used to construct more complex molecules, or they can be broken down into simpler ones. Intermediate metabolites may be synthesized from other metabolites, perhaps used to make more complex substances, or broken down into simpler compounds, often with the release of chemical energy.

[0046] The terms "mevalonate (MV A) pathway" refer to the metabolic pathway which converts acetyl-CoA into dimethylallyl pyrophosphate {DMAPP) and isopentenyl pyrophosphate (IPP) and which is catalyzed by the enzymes acetoacetyl-Coenzyme A synthase, 3-hydroxy-3- methylglutaryl-Coenzyme A synthase, 3-hydroxy-3-methylglutaryl-Coenzyme A reductase, mevalonate kinase (MVK), phosphomevalonate kinase (PMK), diphosphomevalonate decarboxylase (MVD), and isopentenyl diphosphate isomerase (IDI). As used herein the mevalonate pathway can inclade all or a subset of the enzymes of the pathway. For example a subset of enzymes can include acetoacetyl-Coenzyme A synthase, 3-hydroxy-3-methylglutaryl-

Coenzyme A synthase, and 3-hydroxy-3-methylglutaryl-Coenzyme A reductase. In another embodiment, the mevalonate pathway can include mutant enzymes that promote or alter metabolic flux in the pathway.

[0047] A "mutation" means any process or mechanism resulting in a mutant protein, enzyme, polynucleotide, gene, or cell. This includes any mutation in which a protein, enzyme, polynucleotide, or gene sequence is altered, and any detectable change in a cell arising from such a mutation. Typically, a mutation occurs in a polynucleotide or gene sequence, by point mutations, deletions, or insertions of single or multiple nucleotide residues. A mutation includes polynucleotide alterations arising within a protein-encoding region of a gene as well as alterations in regions outside of a protein-encoding sequence, such as, but not limited to, regulatory or promoter sequences. A mutation in a gene can be "silent", i.e., not reflected in an amino acid alteration upon expression, leading to a "sequence-conservative" variant of the gene. This generally arises when one amino acid corresponds to more than one codon. A mutation that gives rise to a different primary sequence of a protein can be referred to as a mutant protein or protein variant.

[0048] A "native" or "wild-type" protein, enzyme, polynucleotide, gene, or cell, means a protein, enzyme, polynucleotide, gene, or cell that occurs in nature.

[0049] As used herein, "non-conservative substitutions” or "non-conservative amino acid exchanges” are defined as exchanges of an amino acid by another amino acid listed in a different groups compared to “conservative substitutions” (see above).

[0050] As used herein a “non-natural amino acid” refers to amino acids that do not occur in nature such as N-methyl amino acids {e.g., N-methyl L-alanine, N-methyl L-valine etc.) or alpha- methyl amino acids, beta-homo amino acids, homo- amino acids and D-amino acids. In a particular embodiment, a non-natural amino acid useful in the disclosure includes a small hydrophobic non-natural amino acid (e.g., N-methyl L-alanine, N-methyl L-valine etc).

[0051] As used herein “operably linked” is meant a functional linkage between two or more domains of a polypeptide or polynucleotide such that the domains function as intended. For example, an expression control sequence (such as a promoter) can be operably linked to a polynucleotide to be expressed, wherein the expression control sequence is operably linked to the polynucleotide such that it directs transcription of the polynucleotide.

[0052] A "parental microorganism" refers to a cell used to generate a recombinant microorganism. The term "parental microorganism” describes, in one embodiment, a cell that occurs in nature, i.e. a "wild-type" cell that has not been genetically modified. The term “parental microorganism" further describes a cell that serves as the “parent” for further engineering. In this latter embodiment, the cell may have been genetically engineered, but serves as a source for further genetic engineering.

[0053] For example, a wild-type microorganism can be genetically modified to express or over express a first target enzyme such as modified HMGR. This microorganism can act as a parental microorganism in the generation of a microorganism modified to express or over-express a second target enzyme e.g., an ERG20. In turn, that microorganism can be modified to express or over express e.g., beta-farnesene synthase. As used herein, “express” or “over express’ refers to the phenotypic expression of a desired gene product. In one embodiment, a naturally occurring gene in the organism can be engineered such that it is linked to a heterologous promoter or regulatory domain, wherein the regulatory domain causes expression of the gene, thereby modifying its normal expression relative to the wild-type organism. Alternatively, the organism can be engineered to remove or reduce a repressor function on the gene, thereby modifying its expression.

In yet another embodiment, a cassette comprising the gene sequence operably linked to a desired expression control/regulatory element is engineered in to the microorganism.

[0054] Accordingly, a parental microorganism functions as a reference cell for successive genetic modification events. Each modification event can be accomplished by introducing one or more nucleic acid molecules into the reference cell. The introduction facilitates the expression or over-expression of one or more target enzyme or the reduction or elimination of one or more target enzymes. It is understood that the term "facilitates" encompasses the activation of endogenous polynucleotides encoding a target enzyme through genetic modification of e.g., a promoter sequence in a parental microorganism. It is further understood that the term "facilitates" encompasses the introduction of exogenous polynucleotides encoding a target enzyme into a parental microorganism.

[0055] Accordingly, metabolically “engineered” or "modified" microorganisms are produced via the introduction of genetic material into a host or parental microorganism of choice thereby modifying or altering the cellular physiology and biochemistry of the microorganism.

Through the introduction of genetic material the parental microorganism acquires new properties, e.g. the ability to produce a new, or greater quantities of, an intracellular metabolite or to express a polypeptide not normally expressed. In an illustrative embodiment, the introduction of genetic material into a parental microorganism results in a new or modified ability to produce sesquiterpene or other terpenoids. The genetic material introduced into the parental microorganism contains gene(s), or parts of gene(s), coding for one or more of the enzymes involved in a biosynthetic pathway for the production of sesquiterpenoids, and may also include additional elements for the expression and/or regulation of expression of these genes, e.g. promoter sequences.

[0056] An engineered or modified microorganism can also include in the alternative or in addition to the introduction of a genetic material into a host or parental microorganism, the disruption, deletion or knocking out of a gene or polynucleotide to alter the cellular physiology and biochemistry of the microorganism. Through the reduction, disruption or knocking out of a gene or polynucleotide the microorganism acquires new or improved properties (e.g., the ability to produce a new or greater quantities of an intracellular metabolite, improve the flux of a metabolite down a desired pathway, and/or reduce the production of undesirable by-products) or eliminates the enzyme from cell free preparations that may compete with a biosynthetic pathway developed from lysed preparations.

[0057] As used herein, a “polynucleotide” or “nucleic acid” refers to two or more deoxyribonucleotides and/or ribonucleotides covalently joined together in either single or double- stranded form. The term includes DNA, RNA, and DNA/RNA hybrid molecules in either single strand our double stranded form.

[0058] It is understood that a polynucleotide described herein include "genes" and that the nucleic acid molecules described above include vectors” or "plasmids."

[0059] Those of skill in the art will recognize that, due to the degenerate nature of the genetic code, a variety of codons differing in their nucleotide sequences can be used to encode a given amino acid. A particular polynucleotide or gene sequence encoding a biosynthetic enzyme or polypeptide described above are referenced herein merely to illustrate an embodiment of the disclosure, and the disclosure includes polynucleotides of any sequence that encode a polypeptide comprising the same amino acid sequence of the polypeptides and proteins of the enzymes utilized in the methods of the disclosure. In similar fashion, a polypeptide can typically tolerate one or more amino acid substitutions, deletions, and insertions in its amino acid sequence without loss or significant loss of a desired activity. The disclosure includes such polypeptides with alternate amino acid sequences, and the amino acid sequences encoded by the DNA sequences shown herein merely illustrate exemplary embodiments of the disclosure.

[0060] The disclosure provides polynucleotides in the form of recombinant DNA expression vectors or plasmids, as described in more detail elsewhere herein, that encode one or more target enzymes. Generally, such vectors can either replicate in the cytoplasm of the host microorganism or integrate into the chromosomal DNA of the host microorganism. In either case,

the vector can be a stable vector (1.e., the vector remains present over many cell divisions, even if only with selective pressure) or a transient vector (i.e, the vector is gradually lost by host microorganisms with increasing numbers of cell divisions). The disclosure provides DNA molecules in isolated (i.e.. not pure, but existing in a preparation in an abundance and/or concentration not found in nature) and purified (i.e, substantially free of contaminating materials or substantially free of materials with which the corresponding DNA would be found in nature) form.

[0061] A polynucleotide of the disclosure can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques and those procedures described in the Examples section below. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by

DNA sequence analysis. Furthermore, oligonucleotides corresponding to nucleotide sequences can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer.

[0062] The disclosure provides a number of polypeptide sequences in the sequence listing accompanying the present application, which can be used to design, synthesize and/or isolate polynucleotide sequences using the degeneracy of the genetic code or using publicly available databases to search for the coding sequences.

[0063] It is also understood that an isolated polynucleotide molecule encoding a polypeptide homologous to the enzymes described herein can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence encoding the particular polypeptide, such that one or more amino acid substitations, additions or deletions are introduced into the encoded protein. Mutations can be introduced into the polynucleotide by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. In contrast to those positions where it may be desirable to make a non-conservative amino acid substitution, in some positions it is preferable to make conservative amino acid substitutions.

[0064] As will be understood by those of skill in the art, it can be advantageous to modify a coding sequence to enhance its expression in a particular host. The genetic code is redundant with 64 possible codons, but most organisms typically use a subset of these codons. The codons that are utilized most often in a species are called optimal codons, and those not utilized very often are classified as rare or low-usage codons. Codons can be substituted to reflect the preferred codon usage of the host, a process sometimes called "codon optimization” or "controlling for species codon bias."

[0065] Optimized coding sequences containing codons preferred by a particular prokaryotic or eukaryotic host (see also, Murray et al. (1989) Nucl. Acids Res. 17:477-508) can be prepared, for example, to increase the rate of translation or to produce recombinant RNA transcripts having desirable properties, such as a longer half-life, as compared with transcripts produced from a non-optimized sequence. Translation stop codons can also be modified to reflect host preference. For example, typical stop codons for S. cerevisiae and mammals are UAA and

UGA, respectively. The typical stop codon for monocotyledonous plants is UGA, whereas insects and E. coli commonly use UAA as the stop codon {Dalphin et al. (1996) Nucl. Acids Res. 24: 216- 218). Methodology for optimizing a nucleotide sequence for expression in a plant is provided, for example, in U.S. Pat. No. 6,015,891, and the references cited therein.

[0066] As used herein, the term "polypeptides" includes polypeptides, proteins, peptides, fragments of polypeptides, and fusion polypeptides.

[0067] The term “substrate” or "suitable substrate” refers to any substance or compound that is converted or meant to be converted into another compound by the action of an enzyme. The term includes not only a single compound, but also combinations of compounds, such as solutions, mixtures and other materials which contain at least one substrate, or derivatives thereof. Further, the term "substrate" encompasses not only compounds that provide a starting material, but also intermediate and end product metabolites used in a pathway associated with a metabolically engineered microorganism as described herein.

[0068] As used herein, "terpenes" are a large and varied class of hydrocarbons that have a simple unifying feature, despite their structural diversity. All terpenes consist of isoprene (C5) units. A terpene classification system depends on the number of such units. Monoterpenes comprise 2 isoprene units and are classified as (C10) terpenes, sesquiterpenes comprise 3 isoprene units and are classified as (C15) terpenes, diterpenes comprise 4 isoprene units and are classified as (C20) terpenes, sesterterpenes (C25), triterpenes (C30) and rubber (C5)... Terpenes occur as acyclic or mono- to pentacyclic derivatives with alcohol, ether, ester, acids, aldehyde, or ketone groups (the so called "terpenoids"), everywhere in organisms, particularly in higher plants, and are characteristic of the individual type of plants. Terpenes such as Monoterpenes (C10),

Sesquiterpenes (C15) and Diterpenes (C20) are derived from the prenyl diphosphate substrates, geranyl diphosphate (GPP), farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) respectively through the action of a very large group of enzymes called the terpene (terpenoid) synthases. These enzymes are often referred to as terpene cyclases since the product of the reactions are cyclized to various monoterpene, sesquiterpene and diterpene carbon skeleton products. Many of the resulting carbon skeletons undergo subsequence oxygenation by cytochrome p450 hydrolysase enzymes to give rise to large families of derivatives. The technical syntheses of top-selling flavors and fragrances can start from terpenes which can also serve as excellent solvents or diluting agents for dyes and varnishes. Natural or synthetic resins of terpenes are used and also many pharmaceutical syntheses of vitamins and insecticides start from terpenes. As used herein, the term "terpene” or "sesquiterpene" (for example) includes corresponding terpenoid or sesquiterpenoid compounds.

[0069] "Transformation" refers to the process by which a vector is introduced into a host cell. Transformation (or transduction, or transfection), can be achieved by any one of a number of means including electroporation, microinjection, biolistics (or particle bombardment-mediated delivery), or agrobacterium mediated transformation.

[0070] In another embodiment, one or more polynucleotides encoding one or more enzymes of the pathway are cloned into one or more microorganism under conditions whereby the enzymes are expressed. Subsequently the cells are lysed and the lysed preparation comprising the one or more enzymes derived from the cell are combined with a suitable buffer and substrate (and one or more additional enzymes of the pathway, if necessary) to produce the prenylated compound or the cannabinoids or cannabinoid precursor. Alternatively, the enzymes can be isolated from the tysed preparations and then recombined in an appropriate buffer. In yet another embodiment, a combination of purchased enzymes and expressed enzymes are used to provide a pathway in an appropriate buffer. In one embodiment, heat stabilized polypeptide/enzymes of the pathway are cloned and expressed. In one embodiment, the enzymes of the pathway are derived from thermophilic microorganisms. The microorganisms are then lysed, the preparation heated to a temperature wherein the heat stabilized polypeptides of the pathway are active and other polypeptides {not of interest) are denatured and become inactive. The preparation thereby includes a subset of all enzymes in the microorganism and includes active heat-stable enzymes. The preparation can then be used to carry out the pathway to produce the prenylated compound or the cannabinoids or cannabinoid precursor.

[0071] For example, to construct an in vitro system, all the enzymes can be acquired commercially or purified by affinity chromatography, tested for activity, and mixed together in a properly selected reaction buffer.

[0072] An in vivo system is also contemplated using all or portions of the foregoing enzymes in a biosynthetic pathway engineered into a microorganism to obtain a recombinant microorganism.

[0073] A “vector” generally refers to a polynucleotide that can be propagated and/or transferred between organisms, cells, or cellular components. Vectors include viruses, bacteriophage, pro-viruses, plasmids, phagemids, transposons, and artificial chromosomes such as

YACs (yeast artificial chromosomes), BACs (bacterial artificial chromosomes), and PLACs {plant artificial chromosomes), and the like, that are "episomes,"” that is, that replicate autonomously or can integrate into a chromosome of a host cell. A vector can also be a naked RNA polynucleotide, a naked DNA polynucleotide, a polynucleotide composed of both DNA and RNA within the same strand, a poly-lysine-conjugated DNA or RNA, a peptide-conjugated DNA or RNA, a liposome- conjugated DNA, or the like, that are not episomal in nature, or it can be an organism which comprises one or more of the above polynucleotide constructs such as an agrobacterium or a bacterium.

[0074] The various components of an expression vector can vary widely, depending on the intended use of the vector and the host cell(s) in which the vector is intended to replicate or drive expression. Expression vector components suitable for the expression of genes and maintenance of vectors in E. coli, yeast, Streptomyces, and other commonly used cells are widely known and commercially available. For example, suitable promoters for inclusion in the expression vectors of the disclosure include those that function in eukaryotic or prokaryotic host microorganisms. Promoters can comprise regulatory sequences that allow for regulation of expression relative to the growth of the host microorganism or that cause the expression of a gene to be turned on or off in response to a chemical or physical stimulus. For E. coli and certain other bacterial host cells, promoters derived from genes for biosynthetic enzymes, antibiotic-resistance conferring enzymes, and phage proteins can be used and include, for example, the galactose, lactose (lac), maltose, tryptophan (trp), beta-lactamase (bla), bacteriophage lambda PL, and T5 promoters. In addition, synthetic promoters, such as the tac promoter (U.S. Pat. No. 4,551,433, which is incorporated herein by reference in its entirety), can also be used. For E. coli expression vectors, it is useful to include an E. coli origin of replication, such as from pUC, plP, pt, and pBR.

[0075] Thus, recombinant expression vectors contain at least one expression system, which, in turn, is composed of at least a portion of a gene coding sequences operably linked to a promoter and optionally termination sequences that operate to effect expression of the coding sequence in compatible host cells. The host cells are modified by transformation with the recombinant DNA expression vectors of the disclosure to contain the expression system sequences either as extrachromosomal elements or integrated into the chromosome.

[0076] The sequences provided herein and the accession numbers provide those of skill in the art the ability to obtain and obtain coding sequences for various enzymes of the disclosure using readily available software and basis biology knowledge.

[0077] The sequence listing appended hereto provide exemplary polynucleotides and polypeptides useful in the methods described herein. It is understood that the addition of sequences which do not alter the activity of a polypeptide or polynucleotide molecule, such as the addition of a non-functional or non-coding sequence (e.g., polyHIS tags), is a conservative variation of the basic molecule.

[0078] Culture conditions suitable for the growth and maintenance of a recombinant microorganism provided herein are known (see, e.g., "Culture of Animal Cells--A Manual of Basic

Technique" by Freshney, Wiley-Liss, N.Y. (1994), Third Edition). The skilled artisan will recognize that such conditions can be modified to accommodate the requirements of each microorganism.

[0079] It is understood that a range of microorganisms can be modified to include all or part of a recombinant metabolic pathway suitable for the production of prenylated compounds or cannabinoids or cannabinoid precursors. It is also understood that various microorganisms can act as "sources" for genetic material encoding target enzymes suitable for use in a recombinant microorganism provided herein.

[0080] General texts which describe molecular biological techniques useful herein, including the use of vectors, promoters and many other relevant topics, include Berger and

Kimmel, Guide to Molecular Cloning Techniques, Methods in Enzymology Volume 152, (Academic Press, Inc., San Diego, Calif.) ("Berger"); Sambrook et al., Molecular Cloning--A

Laboratory Manual, 2d ed., Vol. 1-3, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y, 1989 ("Sambrook") and Current Protocols in Molecular Biology, F. M. Ausubel et al, eds,

Current Protocols, a joint venture between Greene Publishing Associates, Inc. and John Wiley &

Sons, Inc., (supplemented through 1999) ("Ausubel"), each of which is incorporated herein by reference in its entirety.

[0081] Examples of protocols sufficient to direct persons of skill through in vitro amplification methods, including the polymerase chain reaction (PCR), the ligase chain reaction (LCR), Qp-replicase amplification and other RNA polymerase mediated techniques (e.g.,

NASBA), e.g., for the production of the homologous nucleic acids of the disclosure are found in

Berger, Sambrook, and Ausubel, as well as in Mullis et al. (1987) U.S. Pat. No. 4,683,202; Innis et al, eds. (1990) PCR Protocols: A Guide to Methods and Applications (Academic Press Inc. San

Diego, Calif.) ("Innis"); Arnhem & Levinson (Oct. 1, 1990) C&EN 36-47; The Journal Of NIH

Research (1991) 3: 81-94; Kwoh et al. (1989) Proc. Natl. Acad. Sci. USA 86: 1173; Guatelli et al. (1990) Proc. Nat'l. Acad. Sci. USA 87: 1874; Lomell et al. (1989) J. Clin. Chem 35: 1826;

Landegren ef al. (1988) Science 241: 1077-1089; Van Brunt (1990) Biotechnology 8: 291-294; Wu and Wallace (1989) Gene 4:560; Barringer et al. (1990) Gene 89:117; and Sooknanan and Malek (1995) Biotechnology 13:563-564.

[0082] Improved methods for cloning in vitro amplified nucleic acids are described in

Wallace et al., U.S. Pat. No. 5,426,039.

[0083] Improved methods for amplifying large nucleic acids by PCR are summarized in

Cheng et al. (1994) Nature 369: 684-685 and the references cited therein, in which PCR amplicons of up to 40 kb are generated. One of skill will appreciate that essentially any RNA can be converted into a double stranded DNA suitable for restriction digestion, PCR expansion and sequencing using reverse transcriptase and a polymerase. See, e.g., Ausubel, Sambrook and

Berger, all supra.

[0084] The generation of sesquiterpene from FPP would be useful for the production of chemical commodities. Redirection of metabolic flux toward sesquiterpene production requires targeted conversion of FPP (Farnesyl pyrophosphate) to the sesquiterpene of interest through expression of a specific terpene synthase. Historically, sesquiterpene synthases, including 3-

Farnesene synthase, have been difficult to identify by typical homology and Hidden Markov

Models (HMM) derived searching methodologies. The level of homology between deduced amino acid sequences of synthases showing identical substrate and product specificity from different species can be lower than the homology between synthases catalyzing the formation of completely different terpene structural types from the same plant.

[0085] Sesquiterpene synthases are known as the rate-limiting enzyme in many heterologous biosynthetic pathways. High levels of production often requires a high-copy plasmid expression or many genomic copies of the synthase, which can lead to genetic instability of the cell lineage.

Therefore, there is great interest in identifying sesquiterpene synthases with improved kinetics, so that fewer copies of synthase yield similar titers thereby reducing instability and preventing toxic buildup of metabolic intermediates. This disclosure provides several novel -Farnesene synthases enzymes that provide microorganisms (e.g., yeast) with increased flux from FPP to p-Farnesene.

[0086] In the past two decades, genes encoding (E)-p-Farnesene synthases has been isolated from Mentha piperita, Citrus junos, and Zea mays and have been shows to make p-

Farnesene product in vitro. These existing sequences were used as seed sequences to perform a homology based search where these and other novel B-Farnesene synthases were validated for production of B-Farnesene in vivo in recombinant microorganism such as S. cerevisiae.

[0087] In one embodiment, the disclosure provides a recombinant microorganism that metabolizes a substrate such as glucose, acetyl-coA, acetoacetyl-coA or mevalonate to B-

Farnesene. In one embodiment, the microorganism is a yeast, such as S. cerevisiae. In one embodiment, the microorganism comprises enzymes of the mevalonate pathway. In certain embodiments, particular enzymes of the mevalonate pathway are mutated to improve metabolic flux and/or cofactor use.

[0088] The disclosure provides a recombinant metabolic pathway engineered into a microorganism (e.g., S. cerevisiae) or prepared as a cell free system. In one embodiment, the recombinant metabolic pathway coverts two molecules acetyl-coA to acetoacetyl-coA by the action of acetoacetyl-coA thiolase (aka acetyl-coA acetyltransferase). Acetoacetyl-coA thiolase is in E. coli comprises AtoB (the polypeptide is SEQ ID NO: 1). In certain embodiments, the acetyl-coA acetyltransferase is ERG10 from S. cerevisiae (SEQ ID NO:3) or sequences that are 85-99% identical thereto. Acetyl-coA acetyltransferase is encoded in C. acetobutylicum by thiA. THL and

AtoB homologs and variants are known. For examples, such homologs and variants include, for example, acetyl-coA acetyltransferase (thiolase) (Streptomyces coelicolor A3(2)) gi[21224359l[ref|NP--630138.1/(21224359); acetyl-coA acetyltransferase (thiolase) (Streptomyces coelicolor A3(2)) gi|3169041|emb|CAA19239.1|(3169041); Acetyl CoA acetyltransferase

(thiolase) (Alcanivorax borkumensis SK2) gi|1 10834428|ref] YP--693287.1|(110834428); Acetyl

CoA acetyltransferase (thiolase) (Alcanivorax borkumensis SK2) gill 10647539)emb|CAL17015.1|(1 10647539); acetyl CoA acetyltransferase (thiolase) (Saccharopolyspora erythraea NRRL 2338) gi|133915420|emb|CAMO5533. 1|(133915420); acetyl- coA acetyltransferase (thiolase) (Saccharopolyspora erythraea NRRL 2338) gil 134098403 ref] YP-- 001104064.1(134098403); acetyl-coa acetyltransferase (thiolase) (Saccharopolyspora erythraea

NRRL 2338) gi|133911026|emb|CAMO01139.1]|(13391 1026); acetyl-CoA acetyltransferase (thiolase) (Clostridium botulinum A str. ATCC 3502) gijl48290632lembICAL84761.1]{148290632); acetyl-CoA acetyltransterase (thiolase) (Pseudomonas aeruginosa UCBPP-PA14) gi|1 15586808|gb|ABT12823.1|(1 15586808); acetyl-CoA acetyltransferase (thiolase) (Ralstonia metallidurans CH34) gi[93358270|gblABF12358.1|(93358270); acetyl-CoA acetyltransferase (thiolase) (Ralstonia metallidurans CH34) gi|93357190|gb|ABF11278.1|(93357190); acetyl-CoA acetyltransferase (thiolase) (Ralstonia metallidurans CH34) gi|93356587|gb|ABF10675.1|(93356587); acetyl-CoA acetyltransferase (thiolase) (Ralstonia eutropha JMP134) 2i|72121949|gb|AAZ64135.1|(72121949); acetyl-CoA acetyltransferase (thiolase) (Ralstonia eutropha IMP134)g1|72121729| gb|AAZ63915.11(72121729); acetyl-CoA acetyltransferase (thiolase) (Ralstonia eutropha JMP134) gi[72121320|gb|AAZ63506.1[(72121320); acetyl-CoA acetyltransferase (thiolase) (Ralstonia eutropha JMP134) gi[72121001|gblAAZ63187.1|(72121001); acetyl-CoA acetyltransferase (thiolase) (Escherichia coli) gi|2764832|emb|CAA66(99.1|(2764832), each sequence associated with the accession number is incorporated herein by reference in its entirety.

[0089] In some embodiments, the acetyl-coA acetyltransferase is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%. 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 3. In some embodiments, the acetyl-coA acetyltranferase is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 06%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID

NO: 3. In certain embodiments, the acetyl-coA acetyl transferase has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%. 90%, 91%, 92%, 93%, 94%, 95% 96%. 97%, 98%, 99%, or 100%, or any range between any two of the foregoing valaes, identical to the amino acid sequence of SEQ ID NO: 3.

[0090] In some embodiments, the acetyl-coA acetyltransferase comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 3. In some embodiments, the acetyl-coA acetyltransferase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 3. In some embodiments, the acetyl-coA acetyltransferase comprises 1,2,3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 3, In some embodiments, the acetyl-coA acetyltransferase comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 3. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[0091] In another or further embodiment, the recombinant metabolic pathway converts acetoacetyl-coA to HMG-coA by the action of HMGS: hydroxymethyiglutaryl-CoA synthase, EC

Number: 2.3.3.10. In S. cerevisiae HMGS is encoded by ERG13. HMGS enzymes from various species are known in the art. In one embodiment, an HMGS useful in the methods and composition of the disclosure is obtained from S. cerevisiae and comprises a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO:5.

[0092] In some embodiments, the HMGS is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 893%, 90%, 91%, 92%, 93%, 94%, 05% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the HMGS is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at feast 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 5. In certain embodiments, the

HMGS has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%. 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 5.

[0093] In some embodiments, the HMGS comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 5. In some embodiments, the HMGS comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 5. In some embodiments, the HMG2 comprises 1, 2, 3,4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,

20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the HMGS comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 5. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[0094] In another or further embodiment, the recombinant metabolic pathway converts

HMG-coA and NADH to mevalonate and NAD+ by the action of HMGR: hydroxymethylglutaryl-

CoA reductase, EC Number: 1.1.1.34. In S. cerevisiae HMGR is encoded by HMGI. HMGR enzymes from various species are known in the art. In one embodiment, an HMGR useful in the methods and composition of the disclosure is obtained from S. cerevisiae and comprises a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID

NO:7. In another embodiment, the HMGR can be a truncated HMGR that comprises or consists of the catalytic domain of HMGR and which has a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO:8.

[0095] In some embodiments, the HMGR or truncated HMGR is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 7 or 8. In some embodiments, the HMGR or truncated HMGR is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%. at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 7 or 8. In certain embodiments, the HMGR or truncated HMGR has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%. 86%, 87%, 88%. 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 7 or 8.

[0096] In some embodiments, the HMGR or truncated HMGR comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 7 or 8. In some embodiments, the HMGR or truncated HMGR comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 7 or 8. In some embodiments, the HMGR or truncated HMGR comprises 1,2,3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 7 or 8. In some embodiments, the HMGR or truncated HMGR comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 7 or 8. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[0097] In another or further embodiment, the disclosure provides a recombinant metabolic pathway that functions in a microorganism. The recombinant metabolic pathway converts mevalonate and ATP by the action of mevalonate kinase (MVK or ERG12) to produce mevalonate-5-phosphate and ADP, Melavonate kinases are known in the art and include sequence that are at least 85-100% (e.g., 85%, 90%, 95%, 98%, 99%) identical to the sequence of SEQ ID

NO: 10 and which have mevalonate kinase activity. In one embodiment, the MVK is ERG12 having a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ

ID NO:19.

[0098] In some embodiments, the mevalonate kinase is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the mevalonate kinase is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 10. In certain embodiments, the mevalonate kinase has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%. 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 10.

[0099] In some embodiments, the mevalonate kinase comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 10. In some embodiments, the mevalonate kinase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 10. In some embodiments, the mevalonate kinase comprises 1,2, 3,4,5,6,7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 10. In some embodiments, the mevalonate kinase comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, atleast 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 10. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[00100] In another or further embodiment, the mevalonate-5-phosphate is further phosphorylated by ATP and the actions of phosphomevalonate kinase (PMVK or ERGS) to produce mevalonate-5-diphosphate and ADP. Phosphomevalonate kinases are known in the art and include sequence that are at least 85-100% (e.g., 85%, 90%, 95%, 98%, 99%) identical to the sequence of SEQ ID NO: 12 and which have phosphomevalonate kinase activity. In one embodiment, the PMVK is ERGS having a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 12.

[00101] In some embodiments, the phosphomevalonate kinase is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 12. In some embodiments, the phosphomevalonate kinase is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%. at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID

NO: 12. In certain embodiments, the phosphomevalonate kinase has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%. 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%. 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ 1D NO: 12.

[00102] In some embodiments, the phosphomevalonate kinase comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 12. In some embodiments, the phosphomevalonate kinase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 12. In some embodiments, the phosphomevalonate kinase comprises 1, 2, 3,4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of

SEQ ID NO: 12, In some embodiments, the phosphomevalonate kinase comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least

11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 12. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[00103] In another or further embodiment, the mevalonate-5-diphosphate is decarboxylated by ATP and the actions of diphosphomevalonate decarboxylase (MDC or ERG19) to produce

ADP, CO: and isopentyl pyrophosphate (IPP). Diphosphomevalonate decarboxylases are known in the art and include sequence that are at least 85-100% (e.g., 85%, 90%, 95%, 98%. 99%) identical to the sequence of SEQ ID NO: 14 and which have diphosphomevalonate kinase activity.

In one embodiment, the MDC is ERG19 having a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%, 99% or 100% identical to SEQ ID NO: 14.

[00104] In some embodiments, the diphosphomevalonate decarboxylase is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%. 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%. 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the diphosphomevalonate decarboxylase is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 14. In certain embodiments, the diphosphomevalonate decarboxylase has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%. 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 14.

[00105] In some embodiments, the diphosphomevalonate decarboxylase comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 14. In some embodiments, the diphosphomevalonate decarboxylase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 14. In some embodiments, the diphosphomevalonate decarboxylase comprises 1,2, 3,4, 5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 14. In some embodiments, the diphosphomevalonate decarboxylase comprises at least 1, at least 2, at least 3, at least 4, at least 5, atleast 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23,

at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID

NO: 14. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[00106] In another or further embodiment, the pathway allows for the conversion of IPP to dimethylallyl diphosphate (DMAPP) or vice-a-versa using an enzyme having isopentenyl pyrophosphate isomerase (IDI) activity. The isopentenyl pyrophosphate isomerase (IDI), can be a bacterial IDI or yeast IDI. In some embodiments, IDI isomerizes IPP to DMAPP and/or DMAPP to

IPP. While several isopentenyl pyrophosphate isomerases are known, in some embodiments, the isopentenyl pyrophosphate isomerase comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 16 (S. cerevisiae IDI). In some embodiments, the isopentenyl pyrophosphate isomerase is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%. 84%, 85%, 86%, 87%, 88%. 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 16. In some embodiments, the isopentenyl pyrophosphate isomerase is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%. at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 16. In certain embodiments, the isopentenyl pyrophosphate isomerase has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%. 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 16.

[00107] In some embodiments, the isopentenyl pyrophosphate isomerase comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 16. In some embodiments, the isopentenyl pyrophosphate isomerase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 16. In some embodiments, the isopentenyl pyrophosphate isomerase comprises 1, 2,3, 4,5,6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 16. In some embodiments, the isopentenyl pyrophosphate isomerase comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, atleast 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 16.

Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[00108] In another or further embodiment, geranyl pyrophosphate (GPP) is formed from the combination of DMAPP and isopentenyl pyrophosphate (IPP) in the presence of geranyl-PP synthase (GPPS). In some yeast, a GPPS activity resides in a farnesyl diphosphate (or farnesyl pyrophosphate) synthase (FPPS) where it has been suggested that GPP is a precursor of FPP and thus IPP and DMAPP are converted to FPP. S. cerevisiae FPPS is the expression product of the

ERG20 locus. In one embodiment, the FPPS (e.g., ERG20) comprises a sequence that is at least 85%, 87%, 90%, 95%, 97%, 98%. 99% or 100% identical to SEQ ID NO:18.

[00109] In some embodiments, the farnesyl-PP synthase comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 18. In some embodiments, the farnesyl-PP synthase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 18. In some embodiments, the farnesyl-PP synthase comprises 1,2, 3,4, 5, 6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID

NO: 18. In some embodiments, the farnesyl-PP synthase comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, atleast 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 18. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[00110] In some embodiments, the recombinant microorganism comprises multiple copies of an ERG20 coding sequence in the genome.

[00111] In another or further embodiment, the recombinant pathway comprises a farnesene synthase that converts farnesyl diphosphate (FPP) to (E)-p-farnesene. In another or further embodiment of the disclosure, the microorganism comprise a heterologous polynucleotide encoding a farnesene synthase. The disclosure provides a plurality of farnesene synthases useful in the methods and compositions of the disclosure. In one embodiment, the disclosure provides an isolated polypeptide having a sequence that is 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 31%, 92%, 93%, 94%. 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and which has farnesene synthase activity. In some embodiments, the farnesene synthase is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 76%, at least 77%, at least 70%, at least 79%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%,

at least 87%, at least 88%, at least 89%, at least 90%. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ 1D NO: 20, 26, 28, 34, 36, 38 or 42 and which has farnesene synthase activity. In certain embodiments, the farnesene synthase has a sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 76%, 77%, 70%, 79%, 80%, 81%. 82%, 83%, 84%, 85%, 86%, 87%. 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100%, or any range between any two of the foregoing values, identical to the amino acid sequence of SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and which has farnesene synthase activity.

[00112] In some embodiments, a farnesene synthase of the disclosure comprises from 1 to about 20 or from 1 to about 10 amino acid modifications with respect to SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and which has farnesene synthase activity. In some embodiments, the farnesene synthase comprises from 1 to 5 amino acid modifications with respect to SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and has farnesene synthase activity. In some embodiments, the farnesene synthase comprises 1,2,3.4,5,6,7,8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 amino acid modifications with respect to the amino acid sequence of SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and has famesene synthase activity. In some embodiments, the farnesene synthase comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, atleast 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 35, at least 40, or at least 45, amino acid modifications with respect to the amino acid sequence of

SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and has farnesene synthase activity. Amino acid modifications can be independently selected from amino acid substitutions, insertions, and deletions.

[00113] The disclosure also provides isolated polynucleotides encoding a polypeptide having a sequence that is at least 85% identical to a sequence selected from the group consisting of

SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42 wherein the polypeptide has farnesene synthase activity.

In certain embodiments the polynucleotide hybridizes under moderate to highly stringent conditions to a polynucleotide sequence consisting of SEQ ID NO:19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, and wherein the polynucleotide encodes a polypeptide having famesene synthase activity. ln another embodiment, the polynucleotide is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41 and wherein the polynucleotide encodes a polypeptide having farnesene synthase activity. In still another embodiment, the disclosure provides a codon optimized polynucleotide sequence of any one of SEQ ID Nos: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, wherein the polynucleotide is codon optimized for expression in a Saccharomyces sp. In still another embodiment, any polynucleotide sequence can have “T” replaced with “U” (i.e., an RNA sequence).

[00114] The disclosure further provides a vector comprising a polynucleotide sequence of the disclosure {e.g., a sequence having at least 85% identity to SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41 and encoding a polypeptide having farnesene synthase activity).

[00115] “Hybridization” can refer to a reaction in which one or more polynucleotides react to form a complex that is stabilized via hydrogen bonding between the bases of the nucleotide residues. The hydrogen bonding can occur by Watson-Crick base pairing, Hoogstein binding, or in any other sequence-specific manner. The complex can comprise two strands forming a duplex structure, three or more strands forming a malti-stranded complex, a single self-hybridizing strand, or any combination of these. A hybridization reaction can constitute a step in a more extensive process, such as the initiation of a PC reaction, or the enzymatic cleavage of a polynucleotide by a ribozyme.

[00116] Examples of stringent hybridization conditions include: incubation temperatures of about 25°C to about 37°C; hybridization buffer concentrations of about 6x SSC to about 10x SSC; formamide concentrations of about 0% to about 25%; and wash solutions from about 4x SSC to about 8x SSC. Examples of moderate hybridization conditions include: incubation temperatures of about 40°C to about 50°C; buffer concentrations of about 9x SSC to about 2x SSC; formamide concentrations of about 30% to about 50%; and wash solutions of about 5x SSC to about 2x SSC.

Examples of high stringency conditions include: incubation temperatures of about 55°C to about 68°C: buffer concentrations of about Ix SSC to about 0.1x SSC; formamide concentrations of about 55% to about 75%; and wash solutions of about ix SSC, 0.1x SSC, or deionized water. ln general, hybridization incubation times are from 5 minutes to 24 hours, with 1, 2, or more washing steps, and wash incubation times are about 1, 2, or 15 minutes. SSC is 0.15 M NaCl and 15 mM citrate buffer. It is understood that equivalents of SSC using other buffer systems can be employed.

[00117] In one embodiment, the disclosure provides a recombinant microorganism {e.g., a yeast) that expresses a polynucleotide selected from the group consisting of (1) a polynucleotides encoding a polypeptide having a sequence that is at least 85% identical to a sequence selected from the group consisting of SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42 wherein the polypeptide has farnesene synthase activity; (ii) a polynucleotide that hybridizes under moderate to highly stringent conditions to a polynucleotide sequence consisting of SEQ ID NO:19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, and wherein the polynucleotide encodes a polypeptide having farnesene synthase activity (iii) a polynucleotide that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%. 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41 and wherein the polynucleotide encodes a polypeptide having farnesene synthase activity; (iv) a codon optimized polynucleotide sequence of any one of SEQ ID Nos: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, wherein the polynucleotide is codon optimized for expression in a

Saccharomyces sp.; and (v) any of (i)-(1v) wherein “T” is replaced with “U” (i.e., an RNA sequence). In a further embodiment, the microorganism further comprises one or more of the enzymes selected from Acetoacetyl-coA thiolase (acetyl-coA acetyltransferase);

Hydroxymethyglutaryl-CoA synthase (HMGS); Hydroxymethyglutaryl-coA reductase (HMGR);

Mevalonate kinase (ERG12); Phosphomevalonate kinase (ERGS); Diphosphomevalonate decarboxylase (ERG19); Isopentenyl pyrophosphate isomerase (IDI); Farnesyl diphosphate synthase (FPPS; ERG20); and any combination thereof.

[00118] In one embodiment, the disclosure provides a recombinant yeast cell comprising

Acetoacetyl-coA thiolase (acetyl-coA acetyltransferase); Hydroxymethyglutaryl-CoA synthase (HMGS); Hydroxymethyghutaryl-coA reductase (HMGR); Mevalonate kinase (ERG12);

Phosphomevalonate kinase (ERGS): Diphosphomevalonate decarboxylase (ERG19); Isopentenyl pyrophosphate isomerase (1D); Farnesyl diphosphate synthase (FPPS; ERG20); and comprises a polynucleotide a polynucleotide selected from the group consisting of (i) a polynucleotides encoding a polypeptide having a sequence that is at least 85% identical to a sequence selected from the group consisting of SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42 wherein the polypeptide has farnesene synthase activity; (ii) a polynucleotide that hybridizes under moderate to highly stringent conditions to a polynucleotide sequence consisting of SEQ ID NO:19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, and wherein the -polynucleotide encodes a polypeptide having farnesene synthase activity (iii) a polynucleotide that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41 and wherein the polynucleotide encodes a polypeptide having farnesene synthase activity; (iv) a codon optimized polynucleotide sequence of any one of SEQ ID Nos: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, wherein the polynucleotide is codon optimized for expression in a

Saccharomyces sp.; and (v) any of (i)-(iv) wherein “T” is replaced with “U” (i.e., an RNA sequence).

[00119] The disclosure also provide methods of producing (-farnesene, the method comprising culturing a recombinant microorganism that expresses a polynucleotide selected from the group consisting of (i) a polynucleotides encoding a polypeptide having a sequence that is at least 85% identical to a sequence selected from the group consisting of SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42 wherein the polypeptide has farnesene synthase activity; (ii) a polynucleotide that hybridizes under moderate to highly stringent conditions to a polynucleotide sequence consisting of SEQ ID NO:19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, and wherein the polynucleotide encodes a polypeptide having farnesene synthase activity (iii) a polynucleotide that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95% 96%, 97%, 98%, 99%, or 100% identical to a sequence selected from the group consisting of SEQ ID NO: 19, 21, 23,25, 27, 29, 31, 33, 35, 37, 39, or 41 and wherein the polynucleotide encodes a polypeptide having farnesene synthase activity; (iv) a codon optimized polynucleotide sequence of any one of

SEQ ID Nos: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, or 41, wherein the polynucleotide is codon optimized for expression in a Saccharomyces sp.; and (v) any of (i)-(iv) wherein “IT” is replaced with “U” (i.e., an RNA sequence); under conditions and with a suitable substrate, wherein the substrate is metabolized to B-farnesene. In a further embodiment, the microorganism further comprises one or more of the enzymes selected from Acetoacetyl-coA thiolase (acetyl-coA acetyltransferase); Hydroxymethyglutaryl-CoA synthase (HMGS); Hydroxymethyglutaryl-coA reductase (HMGR); Mevalonate kinase (ERG12); Phosphomevalonate kinase (ERGS);

Diphosphomevalonate decarboxylase (ERG19); Isopentenyl pyrophosphate isomerase (IDI);

Farnesyl diphosphate synthase (FPPS; ERG20); and any combination thereof. In still a further embodiment, the substrate is glucose. In another or further embodiment, the microorganism is a yeast. In a particular embodiment, the yeast is Saccharomyces cerevisiae.

[00120] The invention is illustrated in the following examples, which are provided by way of illustration and are not intended to be limiting.

EXAMPLES

[00121] Yeast use the mevalonate-dependent (MEV) pathway to convert acetyl coenzyme A (acetyl-CoA) to IPP {lsopentenyl pyrophosphate). Redirection of metabolic flux toward sesquiterpene production requires targeted conversion of FPP (Farnesyl pyrophosphate) to the sesquiterpene of interest through expression of a specific terpene synthase. Historically sesquiterpene synthases, including (3-Farnesene synthase, have been difficult to identify by typical homology and HMM derived searching methodologies. The level of homology between deduced amino acid sequences of synthases showing identical substrate and product specificity from different species can be lower than the homology between synthases catalyzing the formation of completely different terpene structural types from the same plant.

[00122] A search was performed for famesene synthases that could be used to improve p3- farnesene production. Twelve nucleotide sequences encoding seven farnesene synthases that catalyze the conversion of FPP to (E)-B-Farnesene in Saccharomyces cerevisiae were identified.

When codon optimized and expressed in yeast, these sequences provide for detectable production of f-farnesene. The identified B-farnesene synthases (e.g., CjFS (SEQ ID NO:20), CjFS1 (SEQ ID

NO:24), CJFS_RR1 (SEQ ID NO:22), ZdFS1 (SEQ ID NO:36), SmFS2 (SEQ ID NO:34), and

ZmFS_RRI1 (SEQ ID NO:40); show relative increases in catalytic activity over a copy of the best published farnesene synthase in Saccharomyces cerevisiae (Artemisia annua Genbank No.

AY835398; Picaud 2005).

[00123] Novel enzymes identified include but are not limited to: Citrus junos (CjFS, CjFS1,

CJFS_RR1), Zea mays (ZmFS, ZmFS_RR1), Mentha piperita (MpFS, MPES1, MPES_RRI), Zea diploperennis (ZdFS1), Zea perennis (ZpFS1), Salvia miltiorrhiza (SmFS2), and Matricaria chamomilla var. recutita (McFS3). SmFS2 also showed detectable amounts of a-bergamotene and sesquisabinene by GCMS.

[00124] Data using these newly identified enzymes shows that wild type Saccharomyces cerevisiae strains carrying these heterologous farnesene synthases genes and an additional copy of

ERG20 and truncated HMGR show up to 5x improvement in (E)-5-Farnesene production over comparable AaFS strains as assayed by both targeted LCMS and GCMS methodologies. We have designed and ordered saturation mutagenesis libraries to both CjFS and ZdFS and are currently in the process of testing and identifying improved enzyme variants with better kinetics and specificity for the production of (E)-fi-Farnesene.

[00125] S. cerevisiae were transformed via standard LiAc chemical methodologies using a

Cas12-based system for directed nuclease-guided genomic integration. All putative farnesene synthases (FS) were expressed from the GALSO locus, driven by a GALI promoter and GAT2 terminator.

[00126] Resulting strains were grown at 30C in 96 mid-well plates with 2% glucose defined media (modified from Westfall 2012) for 42 hours, before transfer to 4% galactose defined media for 48 hours.

[00127] For Figure 1, samples were diluted with butanol, and analyzed on a RapidFire SPE-MS system using a triple quadrupole 6470 mass spectrometer. A C4 column was used, with 0.1% formic acid in as mobile phase A, and 90/10/0.01 methanol/water/formic acid in mobile phase

B. A specific famesene transition was used in the positive ionization mode with an APCI source: 205/93 m/z. Data were quantitated with RapidFire integrator, and normalized to on-plate positive and negative control strains.

[00128] For Figure 2, samples were diluted with butanol, and analyzed on an Agilent 1290 LC-

MS system using a triple quadrupole 6470 mass spectrometer. A Phenomenex 1.1 micron Kinetic 2.1 x30 mm C18 column was used, with 0.1% formic acid in as mobile phase A, and 0.1% formic acid in acetonitrile in mobile phase B. A rapid gradient method was run with a dual injector system for a total peak to peak time of 0.36 minutes. Detection was at 220 nm with a diode array detector. Data were quantitated with MassHunter, and normalized to on-plate positive and negative control strains.

[00129] External standard curves of trans-beta farnesene were used for absolute quantitation.

[00130] The best farnesene synthase, CJFS_RR1, was subjected to full site-directed saturation mutagenesis using the Inscripta Onyx platform and screened the resulting mutants in plates as outlined above for Figure 3. Twenty eight different mutants were recovered (Table 1, Figure 4) that were statistically improved over the native synthase sequence. The mutants included V12, E59,

T67, A68, K70, H76, A80, F120, K136, A213, V270, A330, 1336, V350, P374, G469, N471, 1532 and T555.

[00131] Table I: Mutations

Mutations

[00132] Many plants enzymes contain N-terminal trafficking sequences that can be detrimental for folding and localization when expressed in heterologous organisms. Therefore, a series of thirteen N-terminal CjFS_RR1 truncations were tested for improved production. Four of the truncations showed significantly improved farnesene titers (Figure 5) when screened in plates as outlined in Figure 3. Every N-terminal truncation of 22 amino acids or greater resulted in no production of farnesene.

[00133] To make sure these hits were relevant at larger scale, a selection of two mutants [CjFS_RR1(T67P) and CjFS_RR1(K136T)] and one truncation (T20) were used in tanks (bioreactors). All three mutants showed significant improvement in tanks (Figure 6).

[00134] Experimental setup for bioreactors: Cultures were inoculated from glycerol stocks to target an initial cell density of 1*10%6 cells in the first seed. 0.5 mL of the culture from the first seed was diluted into a second seed flask containing 50 mL minimal media at 0.375 mg/mL in a 250 mL baffled flask and incubated for 24 hours. Cultivation conditions for both seed flasks were as follows: temperature was controlled at 30 °C, agitation at 225 rpm in a 1 inch throw shaker to allow enough oxygenation. Seed media was previously published by Westfall et al. 10.1073/pnas. 1110740109.

[00135] Three 250 mL bioreactors were inoculated for each strain. Prior to inoculation, each bioreactor was batched with 90 mL of minimal media, 11 mL of isopropyl myristate as an organic overlay to allow product partitioning. Culture from the end of seed 2 was used as an inoculum for each bioreactor with target OD of 0.75 at inoculation time. The temperature in the reactor was controlled at 30 C and pH at 5.0 using 14% ammonium hydroxide. The dissolved oxygen was controlled at 30% by cascading agitation (from 600-2500 rpm) and then airflow (0.5-1 vvm). The initial glucose concentration in the batch phase is 19.5 g/L. The end of batch phase was evident by the second DO spike which coincides with exhaustion of batch glucose along with any produced ethanol. After the batch phase, an exponential feed phase started. The starting feed rate was 0.01185 mL/min, with a volame basis of 100 mL initial volume. The growth rate of the feed was 0.2 /hr, and the exponential feed continued for 4.5 or until OUR was >150 mmol/L/h. Once either of these triggers ends exponential feed, we turned on a constant low flow feed of 1 g_glucose/

L_current_volume/hr. At the same time, we turned on DO spike bolus feeding. The boluses feed 10 g_slucose/L_current_volume over 30 minutes, so they feed at a rate of 20 g_glucose/L._current_volume/hr during that time. Both feeds are based on current volumes in the bioreactors. The feed contained 585 g/L dextrose, 4.5 g/L. potassium phosphate monobasic, 5.12 g/L magnesium sulfate heptahydrate, and 35 mL sulfate solution. Main batch fermentation media was previously published by Westfall et al. 10.1073/pnas. 1110740109.

SEQLTXT

SEQUENCE LISTING

<118> Sestina Bio, LLC <120> HETEROLOGOUS EXPRESSION OF ENZYMES FOR INCREASED B-FARNESENE

PRODUCTION

<130> 00164-002P01 <140> P185819NL00 <141> 2021-10-25 <160> 42 <170> PatentIn version 3.5 <210> 1 <211> 394 <212> PRT <213> Escherichia coli <400> 1

Met Lys Asn Cys Val Ile Val Ser Ala Val Arg Thr Ala Ile Gly Ser 1 5 10 15

Phe Asn Gly Ser Leu Ala Ser Thr Ser Ala Ile Asp Leu Gly Ala Thr

Val Ile Lys Ala Ala Ile Glu Arg Ala Lys Ile Asp Ser Gln His Val

Asp Glu Val Ile Met Gly Asn Val Leu Gln Ala Gly Leu Gly Gln Asn 60

Pro Ala Arg Gln Ala Leu Leu Lys Ser Gly Leu Ala Glu Thr Val Cys 65 70 75 80

Gly Phe Thr Val Asn Lys Val Cys Gly Ser Gly Leu Lys Ser Val Ala 85 90 95

Leu Ala Ala Gln Ala Ile Gln Ala Gly Gln Ala Gln Ser Ile Val Ala 100 105 110

Gly Gly Met Glu Asn Met Ser Leu Ala Pro Tyr Leu Leu Asp Ala Lys

Pagina 1

SEQLTXT

115 120 125

Ala Arg Ser Gly Tyr Arg Leu Gly Asp Gly Gln Val Tyr Asp Val Ile 130 135 140

Leu Arg Asp Gly Leu Met Cys Ala Thr His Gly Tyr His Met Gly Ile 145 150 155 160

Thr Ala Glu Asn Val Ala Lys Glu Tyr Gly Ile Thr Arg Glu Met Gln 165 170 175

Asp Glu Leu Ala Leu His Ser Gln Arg Lys Ala Ala Ala Ala Ile Glu 180 185 190

Ser Gly Ala Phe Thr Ala Glu Ile Val Pro Val Asn Val Val Thr Arg 195 200 205

Lys Lys Thr Phe Val Phe Ser Gln Asp Glu Phe Pro Lys Ala Asn Ser 210 215 220

Thr Ala Glu Ala Leu Gly Ala Leu Arg Pro Ala Phe Asp Lys Ala Gly 225 230 235 240

Thr Val Thr Ala Gly Asn Ala Ser Gly Ile Asn Asp Gly Ala Ala Ala 245 250 255

Leu Val Ile Met Glu Glu Ser Ala Ala Leu Ala Ala Gly Leu Thr Pro 260 265 270

Leu Ala Arg Ile Lys Ser Tyr Ala Ser Gly Gly Val Pro Pro Ala Leu 275 280 285

Met Gly Met Gly Pro Val Pro Ala Thr Gln Lys Ala Leu Gln Leu Ala 290 295 300

Gly Leu Gln Leu Ala Asp Ile Asp Leu Ile Glu Ala Asn Glu Ala Phe 305 310 315 320

Ala Ala Gln Phe Leu Ala Val Gly Lys Asn Leu Gly Phe Asp Ser Glu

Pagina 2

SEQLTXT

325 330 335

Lys Val Asn Val Asn Gly Gly Ala Ile Ala Leu Gly His Pro Ile Gly 340 345 350

Ala Ser Gly Ala Arg Ile Leu Val Thr Leu Leu His Ala Met Gln Ala 355 360 365

Arg Asp Lys Thr Leu Gly Leu Ala Thr Leu Cys Ile Gly Gly Gly Gln 370 375 380

Gly Val Ala Met Val Ile Glu Arg Leu Asn 385 390 <2105 2 <211> 1197 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(1197) <400> 2 atg tct cag aac gtt tac att gta tcg act gcc aga acc cca att ggt 48

Met Ser Gln Asn Val Tyr Ile Val Ser Thr Ala Arg Thr Pro Ile Gly 1 5 10 15 tca ttc cag ggt tct cta tcc tcc aag aca gca gtg gaa ttg ggt gct 96

Ser Phe Gln Gly Ser Leu Ser Ser Lys Thr Ala Val Glu Leu Gly Ala gtt gct tta aaa ggc gcc ttg gct aag gtt cca gaa ttg gat gca tcc 144

Val Ala Leu Lys Gly Ala Leu Ala Lys Val Pro Glu Leu Asp Ala Ser aag gat ttt gac gaa att att ttt ggt aac gtt ctt tct gcc aat ttg 192

Lys Asp Phe Asp Glu Ile Ile Phe Gly Asn Val Leu Ser Ala Asn Leu 60 ggc caa gct ccg gcc aga caa gtt get ttg get gcc ggt ttg agt aat 240

Gly Gln Ala Pro Ala Arg Gln Val Ala Leu Ala Ala Gly Leu Ser Asn 65 70 75 80 cat atc gtt gca agc aca gtt aac aag gtc tgt gca tcc gct atg aag 288

His Ile Val Ala Ser Thr Val Asn Lys Val Cys Ala Ser Ala Met Lys

Pagina 3

SEQLTXT

85 90 95 gca atc att ttg ggt gct caa tcc atc aaa tgt ggt aat gct gat gtt 336

Ala Ile Ile Leu Gly Ala Gln Ser Ile Lys Cys Gly Asn Ala Asp Val 100 105 110 gtc gta gct ggt ggt tgt gaa tct atg act aac gca cca tac tac atg 384

Val Val Ala Gly Gly Cys Glu Ser Met Thr Asn Ala Pro Tyr Tyr Met 115 120 125 cca gca gcc cgt gcg ggt gcc aaa ttt ggc caa act gtt ctt gtt gat 432

Pro Ala Ala Arg Ala Gly Ala Lys Phe Gly Gln Thr Val Leu Val Asp 130 135 140 get gtc gaa aga gat ggg ttg aac gat gcg tac gat ggt cta gcc atg 480

Gly Val Glu Arg Asp Gly Leu Asn Asp Ala Tyr Asp Gly Leu Ala Met 145 150 155 160 ggt gta cac gca gaa aag tgt gcc cgt gat tgg gat att act aga gaa 528

Gly Val His Ala Glu Lys Cys Ala Arg Asp Trp Asp Ile Thr Arg Glu 165 170 175 caa caa gac aat ttt gcc atc gaa tcc tac caa aaa tct caa aaa tct 576

Gln Gln Asp Asn Phe Ala Ile Glu Ser Tyr Gln Lys Ser Gln Lys Ser 180 185 190 caa aag gaa ggt aaa ttc gac aat gaa att gta cct gtt acc att aag 624

Gln Lys Glu Gly Lys Phe Asp Asn Glu Ile Val Pro Val Thr Ile Lys 195 200 205 gga ttt aga ggt aag cct gat act caa gtc acg aag gac gag gaa cct 672

Gly Phe Arg Gly Lys Pro Asp Thr Gln Val Thr Lys Asp Glu Glu Pro 210 215 220 gct aga tta cac gtt gaa aaa ttg aga tct gca agg act gtt ttc caa 720

Ala Arg Leu His Val Glu Lys Leu Arg Ser Ala Arg Thr Val Phe Gln 225 230 235 240 aaa gaa aac ggt act gtt act gcc gct aac gct tct cca atc aac gat 768

Lys Glu Asn Gly Thr Val Thr Ala Ala Asn Ala Ser Pro Ile Asn Asp 245 250 255 ggt gct gca gcc gtc atc ttg gtt tcc gaa aaa gtt ttg aag gaa aag 816

Gly Ala Ala Ala Val Ile Leu Val Ser Glu Lys Val Leu Lys Glu Lys 260 265 270 aat ttg aag cct ttg gct att atc aaa ggt tgg ggt gag gcc gct cat 864

Asn Leu Lys Pro Leu Ala Ile Ile Lys Gly Trp Gly Glu Ala Ala His 275 280 285 caa cca gct gat ttt aca tgg gct cca tct ctt gca gtt cca aag gct 912

Gln Pro Ala Asp Phe Thr Trp Ala Pro Ser Leu Ala Val Pro Lys Ala

Pagina 4

SEQLTXT

290 295 300 ttg aaa cat gct ggc atc gaa gac atc aat tct gtt gat tac ttt gaa 960

Leu Lys His Ala Gly Ile Glu Asp Ile Asn Ser Val Asp Tyr Phe Glu 305 310 315 320 ttc aat gaa gcc ttt tcg gtt gtc ggt ttg gtg aac act aag att ttg 1008

Phe Asn Glu Ala Phe Ser Val Val Gly Leu Val Asn Thr Lys Ile Leu 325 330 335 aag cta gac cca tct aag gtt aat gta tat ggt ggt gct gtt gct cta 1056

Lys Leu Asp Pro Ser Lys Val Asn Val Tyr Gly Gly Ala Val Ala Leu 340 345 350 ggt cac cca ttg ggt tgt tct ggt gct aga gtg gtt gtt aca ctg cta 1104

Gly His Pro Leu Gly Cys Ser Gly Ala Arg Val Val Val Thr Leu Leu 355 360 365 tcc atc tta cag caa gaa gga ggt aag atc ggt gtt gcc gcc att tgt 1152

Ser Ile Leu Gln Gln Glu Gly Gly Lys Ile Gly Val Ala Ala Ile Cys 370 375 380 aat ggt ggt ggt ggt gct tcc tct att gtc att gaa aag ata tga 1197

Asn Gly Gly Gly Gly Ala Ser Ser Ile Val Ile Glu Lys Ile 385 390 395 <2105 3 <211> 398 <212> PRT <213> Saccharomyces cerevisiae <400> 3

Met Ser Gln Asn Val Tyr Ile Val Ser Thr Ala Arg Thr Pro Ile Gly 1 5 10 15

Ser Phe Gln Gly Ser Leu Ser Ser Lys Thr Ala Val Glu Leu Gly Ala

Val Ala Leu Lys Gly Ala Leu Ala Lys Val Pro Glu Leu Asp Ala Ser

Lys Asp Phe Asp Glu Ile Ile Phe Gly Asn Val Leu Ser Ala Asn Leu 60

Gly Gln Ala Pro Ala Arg Gln Val Ala Leu Ala Ala Gly Leu Ser Asn 65 70 75 80

Pagina 5

SEQLTXT

His Ile Val Ala Ser Thr Val Asn Lys Val Cys Ala Ser Ala Met Lys 85 90 95

Ala Ile Ile Leu Gly Ala Gln Ser Ile Lys Cys Gly Asn Ala Asp Val 100 105 110

Val Val Ala Gly Gly Cys Glu Ser Met Thr Asn Ala Pro Tyr Tyr Met 115 120 125

Pro Ala Ala Arg Ala Gly Ala Lys Phe Gly Gln Thr Val Leu Val Asp 130 135 140

Gly Val Glu Arg Asp Gly Leu Asn Asp Ala Tyr Asp Gly Leu Ala Met 145 150 155 160

Gly Val His Ala Glu Lys Cys Ala Arg Asp Trp Asp Ile Thr Arg Glu 165 170 175

Gln Gln Asp Asn Phe Ala Ile Glu Ser Tyr Gln Lys Ser Gln Lys Ser 180 185 190

Gln Lys Glu Gly Lys Phe Asp Asn Glu Ile Val Pro Val Thr Ile Lys 195 200 205

Gly Phe Arg Gly Lys Pro Asp Thr Gln Val Thr Lys Asp Glu Glu Pro 210 215 220

Ala Arg Leu His Val Glu Lys Leu Arg Ser Ala Arg Thr Val Phe Gln 225 230 235 240

Lys Glu Asn Gly Thr Val Thr Ala Ala Asn Ala Ser Pro Ile Asn Asp 245 250 255

Gly Ala Ala Ala Val Ile Leu Val Ser Glu Lys Val Leu Lys Glu Lys 260 265 270

Asn Leu Lys Pro Leu Ala Ile Ile Lys Gly Trp Gly Glu Ala Ala His 275 280 285

Pagina 6

SEQLTXT

Gln Pro Ala Asp Phe Thr Trp Ala Pro Ser Leu Ala Val Pro Lys Ala 290 295 300

Leu Lys His Ala Gly Ile Glu Asp Ile Asn Ser Val Asp Tyr Phe Glu 305 310 315 320

Phe Asn Glu Ala Phe Ser Val Val Gly Leu Val Asn Thr Lys Ile Leu 325 330 335

Lys Leu Asp Pro Ser Lys Val Asn Val Tyr Gly Gly Ala Val Ala Leu 340 345 350

Gly His Pro Leu Gly Cys Ser Gly Ala Arg Val Val Val Thr Leu Leu 355 360 365

Ser Ile Leu Gln Gln Glu Gly Gly Lys Ile Gly Val Ala Ala Ile Cys 370 375 380

Asn Gly Gly Gly Gly Ala Ser Ser Ile Val Ile Glu Lys Ile 385 390 395 <2105 4 <211> 1821 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(1821) <400> 4 atg ggt tcc tcc cca gaa gac gaa att caa cag ttg gaa gaa gaa att 48

Met Gly Ser Ser Pro Glu Asp Glu Ile Gln Gln Leu Glu Glu Glu Ile 1 5 10 15 gcc caa tta gaa caa aag aac gct gct ttg aaa gaa aag aac caa gcc 96

Ala Gln Leu Glu Gln Lys Asn Ala Ala Leu Lys Glu Lys Asn Gln Ala ttg aag tac ggt tcc ggt cgt atg aag caa ttg gaa gac aag att gaa 144

Leu Lys Tyr Gly Ser Gly Arg Met Lys Gln Leu Glu Asp Lys Ile Glu

Pagina 7

SEQLTXT gaa cta cta tct aag atc tac cat ttg gaa aac gaa atc gcc aga ttg 192

Glu Leu Leu Ser Lys Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu 60 aag aag ttg atc ggt gaa cgt tcc ggt tcc cca gaa gat aag atc gcc 240

Lys Lys Leu Ile Gly Glu Arg Ser Gly Ser Pro Glu Asp Lys Ile Ala 65 70 75 80 caa ttg aaa caa aag att caa gct ttg aaa caa gaa aac caa caa cta 288

Gln Leu Lys Gln Lys Ile Gln Ala Leu Lys Gln Glu Asn Gln Gln Leu 85 90 95 gaa gaa gaa aac gcc gct cta gaa tac ggt ggt tct ggt ggt tct ggt 336

Glu Glu Glu Asn Ala Ala Leu Glu Tyr Gly Gly Ser Gly Gly Ser Gly 100 105 110 tct ggt gga tct aaa ttg tct acc aaa ttg tgt tgg tgt ggt atc aag 384

Ser Gly Gly Ser Lys Leu Ser Thr Lys Leu Cys Trp Cys Gly Ile Lys 115 120 125 ggt aga tta aga cca caa aag caa caa caa ttg cac aac acc aac ttg 432

Gly Arg Leu Arg Pro Gln Lys Gln Gln Gln Leu His Asn Thr Asn Leu 130 135 140 caa atg act gaa ttg aag aag caa aag act gct gaa caa aag acc aga 480

Gln Met Thr Glu Leu Lys Lys Gln Lys Thr Ala Glu Gln Lys Thr Arg 145 150 155 160 cct caa aat gtc ggt att aag ggt atc caa atc tac att cca act caa 528

Pro Gln Asn Val Gly Ile Lys Gly Ile Gln Ile Tyr Ile Pro Thr Gln 165 170 175 tgt gtc aac caa tct gaa ttg gaa aag ttc gac ggt gtc tct caa ggt 576

Cys Val Asn Gln Ser Glu Leu Glu Lys Phe Asp Gly Val Ser Gln Gly 180 185 190 aaa tac acc att ggt ttg ggt caa acc aac atg tcc ttt gtt aac gat 624

Lys Tyr Thr Ile Gly Leu Gly Gln Thr Asn Met Ser Phe Val Asn Asp 195 200 205 cgt gaa gac atc tac tcc atg tct ttg act gtt ttg tcc aag ttg atc 672

Arg Glu Asp Ile Tyr Ser Met Ser Leu Thr Val Leu Ser Lys Leu Ile 210 215 220 aaa tcc tac aac att gac acc aac aag atc ggt cgt ttg gaa gtc ggt 720

Lys Ser Tyr Asn Ile Asp Thr Asn Lys Ile Gly Arg Leu Glu Val Gly 225 230 235 240 act gaa act ttg att gac aag tcc aag tct gtc aaa tcc gtc ttg atg 768

Thr Glu Thr Leu Ile Asp Lys Ser Lys Ser Val Lys Ser Val Leu Met 245 250 255

Pagina 8

SEQLTXT caa tta ttc ggt gaa aac acc gat gtc gaa ggt att gac act ttg aac 816

Gln Leu Phe Gly Glu Asn Thr Asp Val Glu Gly Ile Asp Thr Leu Asn 260 265 270 gct tgt tac ggt ggt acc aac gct tta ttc aac tct ttg aac tgg atc 864

Ala Cys Tyr Gly Gly Thr Asn Ala Leu Phe Asn Ser Leu Asn Trp Ile 275 280 285 gaa tcc aac gct tgg gac ggt aga gat gcc att gtc gtt tgt ggt gac 912

Glu Ser Asn Ala Trp Asp Gly Arg Asp Ala Ile Val Val Cys Gly Asp 290 295 300 att gct atc tac gac aag ggt gct gcc aga cca act ggt ggt gct ggt 960

Ile Ala Ile Tyr Asp Lys Gly Ala Ala Arg Pro Thr Gly Gly Ala Gly 305 310 315 320 act gtt gcc atg tgg atc ggt cca gat gct cca att gtt ttc gat tcc 1008

Thr Val Ala Met Trp Ile Gly Pro Asp Ala Pro Ile Val Phe Asp Ser 325 330 335 gtc aga gct tct tac atg gaa cac gct tac gat ttc tac aag cca gac 1056

Val Arg Ala Ser Tyr Met Glu His Ala Tyr Asp Phe Tyr Lys Pro Asp 340 345 350 ttc acc tct gaa tac cca tac gtt gac ggt cac ttc tcc ttg acc tgt 1104

Phe Thr Ser Glu Tyr Pro Tyr Val Asp Gly His Phe Ser Leu Thr Cys 355 360 365 tac gtc aag gct ttg gac caa gtc tac aag tct tac tcc aag aag gct 1152

Tyr Val Lys Ala Leu Asp Gln Val Tyr Lys Ser Tyr Ser Lys Lys Ala 370 375 380 atc tcc aag ggt ttg gtt tct gac cca gct ggt tct gat gct ttg aat 1200

Ile Ser Lys Gly Leu Val Ser Asp Pro Ala Gly Ser Asp Ala Leu Asn 385 390 395 400 gtc ttg aag tat ttc gac tac aac gtt ttc cac gtt cca acc tgt aaa 1248

Val Leu Lys Tyr Phe Asp Tyr Asn Val Phe His Val Pro Thr Cys Lys 405 410 415 ttg gtt act aag tct tac ggt aga tta cta tac aac gat ttc aga gct 1296

Leu Val Thr Lys Ser Tyr Gly Arg Leu Leu Tyr Asn Asp Phe Arg Ala 420 425 430 aac cca caa tta ttc cca gaa gtc gat gct gaa ttg gct acc aga gac 1344

Asn Pro Gln Leu Phe Pro Glu Val Asp Ala Glu Leu Ala Thr Arg Asp 435 440 445 tac gac gaa tcc ttg act gac aag aac att gaa aag acc ttt gtc aac 1392

Tyr Asp Glu Ser Leu Thr Asp Lys Asn Ile Glu Lys Thr Phe Val Asn 450 455 460

Pagina 9

SEQLTXT gtt gcc aag cca ttc cac aag gaa cgt gtt gct caa tct ttg att gtt 14409

Val Ala Lys Pro Phe His Lys Glu Arg Val Ala Gln Ser Leu Ile Val 465 470 475 480 cca acc aac act ggt aac atg tac act gct tct gtt tac gcc gct ttc 1488

Pro Thr Asn Thr Gly Asn Met Tyr Thr Ala Ser Val Tyr Ala Ala Phe 485 490 495 gct tct ttg ttg aac tac gtt ggt tcc gat gac ttg caa ggt aag aga 1536

Ala Ser Leu Leu Asn Tyr Val Gly Ser Asp Asp Leu Gln Gly Lys Arg 500 505 510 gtt ggt tta ttc tct tac ggt tct ggt ttg gct gcc tct cta tac tct 1584

Val Gly Leu Phe Ser Tyr Gly Ser Gly Leu Ala Ala Ser Leu Tyr Ser 515 520 525 tgt aag atc gtt ggt gat gtt caa cac atc atc aag gaa ttg gat atc 1632

Cys Lys Ile Val Gly Asp Val Gln His Ile Ile Lys Glu Leu Asp Ile 530 535 540 acc aac aaa ttg gcc aag aga atc act gaa act cca aag gac tac gaa 1680

Thr Asn Lys Leu Ala Lys Arg Ile Thr Glu Thr Pro Lys Asp Tyr Glu 545 550 555 560 gct gcc att gaa ttg aga gaa aac gct cat ttg aag aag aac ttc aag 1728

Ala Ala Ile Glu Leu Arg Glu Asn Ala His Leu Lys Lys Asn Phe Lys 565 570 575 cca caa ggt tcc atc gaa cac ttg caa tct ggt gtt tac tac ttg acc 1776

Pro Gln Gly Ser Ile Glu His Leu Gln Ser Gly Val Tyr Tyr Leu Thr 580 585 590 aac atc gac gac aaa ttc aga aga tct tac gat gtc aag aag taa 1821

Asn Ile Asp Asp Lys Phe Arg Arg Ser Tyr Asp Val Lys Lys 595 600 605 <210> 5 <211> 606 <212> PRT <213> Saccharomyces cerevisiae <400> 5

Met Gly Ser Ser Pro Glu Asp Glu Ile Gln Gln Leu Glu Glu Glu Ile 1 5 10 15

Ala Gln Leu Glu Gln Lys Asn Ala Ala Leu Lys Glu Lys Asn Gln Ala

Pagina 10

SEQLTXT

Leu Lys Tyr Gly Ser Gly Arg Met Lys Gln Leu Glu Asp Lys Ile Glu

Glu Leu Leu Ser Lys Ile Tyr His Leu Glu Asn Glu Ile Ala Arg Leu 60

Lys Lys Leu Ile Gly Glu Arg Ser Gly Ser Pro Glu Asp Lys Ile Ala 65 70 75 80

Gln Leu Lys Gln Lys Ile Gln Ala Leu Lys Gln Glu Asn Gln Gln Leu 85 90 95

Glu Glu Glu Asn Ala Ala Leu Glu Tyr Gly Gly Ser Gly Gly Ser Gly 100 105 110

Ser Gly Gly Ser Lys Leu Ser Thr Lys Leu Cys Trp Cys Gly Ile Lys 115 120 125

Gly Arg Leu Arg Pro Gln Lys Gln Gln Gln Leu His Asn Thr Asn Leu 130 135 140

Gln Met Thr Glu Leu Lys Lys Gln Lys Thr Ala Glu Gln Lys Thr Arg 145 150 155 160

Pro Gln Asn Val Gly Ile Lys Gly Ile Gln Ile Tyr Ile Pro Thr Gln 165 170 175

Cys Val Asn Gln Ser Glu Leu Glu Lys Phe Asp Gly Val Ser Gln Gly 180 185 190

Lys Tyr Thr Ile Gly Leu Gly Gln Thr Asn Met Ser Phe Val Asn Asp 195 200 205

Arg Glu Asp Ile Tyr Ser Met Ser Leu Thr Val Leu Ser Lys Leu Ile 210 215 220

Lys Ser Tyr Asn Ile Asp Thr Asn Lys Ile Gly Arg Leu Glu Val Gly 225 230 235 240

Pagina 11

SEQLTXT

Thr Glu Thr Leu Ile Asp Lys Ser Lys Ser Val Lys Ser Val Leu Met 245 250 255

Gln Leu Phe Gly Glu Asn Thr Asp Val Glu Gly Ile Asp Thr Leu Asn 260 265 270

Ala Cys Tyr Gly Gly Thr Asn Ala Leu Phe Asn Ser Leu Asn Trp Ile 275 280 285

Glu Ser Asn Ala Trp Asp Gly Arg Asp Ala Ile Val Val Cys Gly Asp 290 295 300

Ile Ala Ile Tyr Asp Lys Gly Ala Ala Arg Pro Thr Gly Gly Ala Gly 305 310 315 320

Thr Val Ala Met Trp Ile Gly Pro Asp Ala Pro Ile Val Phe Asp Ser 325 330 335

Val Arg Ala Ser Tyr Met Glu His Ala Tyr Asp Phe Tyr Lys Pro Asp 340 345 350

Phe Thr Ser Glu Tyr Pro Tyr Val Asp Gly His Phe Ser Leu Thr Cys 355 360 365

Tyr Val Lys Ala Leu Asp Gln Val Tyr Lys Ser Tyr Ser Lys Lys Ala 370 375 380

Ile Ser Lys Gly Leu Val Ser Asp Pro Ala Gly Ser Asp Ala Leu Asn 385 390 395 400

Val Leu Lys Tyr Phe Asp Tyr Asn Val Phe His Val Pro Thr Cys Lys 405 410 415

Leu Val Thr Lys Ser Tyr Gly Arg Leu Leu Tyr Asn Asp Phe Arg Ala 420 425 430

Asn Pro Gln Leu Phe Pro Glu Val Asp Ala Glu Leu Ala Thr Arg Asp 435 440 445

Pagina 12

SEQLTXT

Tyr Asp Glu Ser Leu Thr Asp Lys Asn Ile Glu Lys Thr Phe Val Asn 450 455 460

Val Ala Lys Pro Phe His Lys Glu Arg Val Ala Gln Ser Leu Ile Val 465 470 475 480

Pro Thr Asn Thr Gly Asn Met Tyr Thr Ala Ser Val Tyr Ala Ala Phe 485 490 495

Ala Ser Leu Leu Asn Tyr Val Gly Ser Asp Asp Leu Gln Gly Lys Arg 500 505 510

Val Gly Leu Phe Ser Tyr Gly Ser Gly Leu Ala Ala Ser Leu Tyr Ser 515 520 525

Cys Lys Ile Val Gly Asp Val Gln His Ile Ile Lys Glu Leu Asp Ile 530 535 540

Thr Asn Lys Leu Ala Lys Arg Ile Thr Glu Thr Pro Lys Asp Tyr Glu 545 550 555 560

Ala Ala Ile Glu Leu Arg Glu Asn Ala His Leu Lys Lys Asn Phe Lys 565 570 575

Pro Gln Gly Ser Ile Glu His Leu Gln Ser Gly Val Tyr Tyr Leu Thr 580 585 590

Asn Ile Asp Asp Lys Phe Arg Arg Ser Tyr Asp Val Lys Lys 595 600 605 <210> 6 <211> 3165 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(3165) <400> 6

Pagina 13

SEQLTXT atg ccg ctg cta ttc aag gga ctg aaa cag atg gca aag cca att gcc 48

Met Pro Leu Leu Phe Lys Gly Leu Lys Gln Met Ala Lys Pro Ile Ala 1 5 10 15 tat gtt tca aga ttt tcg gcg aaa cga cca att cat ata ata ctt ttt 96

Tyr Val Ser Arg Phe Ser Ala Lys Arg Pro Ile His Ile Ile Leu Phe tct cta atc ata tcc gca ttc gct tat cta tcc gtc att cag tat tac 144

Ser Leu Ile Ile Ser Ala Phe Ala Tyr Leu Ser Val Ile Gln Tyr Tyr ttc aat ggt tgg caa cta gat tca aat agt gtt ttt gaa act gct cca 192

Phe Asn Gly Trp Gln Leu Asp Ser Asn Ser Val Phe Glu Thr Ala Pro 60 aat aaa gac tcc aac act cta ttt caa gaa tgt tcc cat tac tac aga 240

Asn Lys Asp Ser Asn Thr Leu Phe Gln Glu Cys Ser His Tyr Tyr Arg 65 70 75 80 gat tcc tct cta gat ggt tgg gta tca atc acc gcg cat gaa gct agt 288

Asp Ser Ser Leu Asp Gly Trp Val Ser Ile Thr Ala His Glu Ala Ser 85 90 95 gag tta cca gcc cca cac cat tac tat cta tta aac ctg aac ttc aat 336

Glu Leu Pro Ala Pro His His Tyr Tyr Leu Leu Asn Leu Asn Phe Asn 100 105 110 agt cct aat gaa act gac tcc att cca gaa cta gct aac acg gtt ttt 384

Ser Pro Asn Glu Thr Asp Ser Ile Pro Glu Leu Ala Asn Thr Val Phe 115 120 125 gag aaa gat aat aca aaa tat att ctg caa gaa gat ctc agc gtt tcc 432

Glu Lys Asp Asn Thr Lys Tyr Ile Leu Gln Glu Asp Leu Ser Val Ser 130 135 140 aaa gaa att tct tct act gat gga acg aaa tgg agg tta aga agt gac 480

Lys Glu Ile Ser Ser Thr Asp Gly Thr Lys Trp Arg Leu Arg Ser Asp 145 150 155 160 aga aaa agt ctt ttc gac gta aag acg tta gca tat tct ctc tac gat 528

Arg Lys Ser Leu Phe Asp Val Lys Thr Leu Ala Tyr Ser Leu Tyr Asp 165 170 175 gta ttt tca gaa aat gta acc caa gca gac ccg ttt gac gtc ctt att 576

Val Phe Ser Glu Asn Val Thr Gln Ala Asp Pro Phe Asp Val Leu Ile 180 185 190 atg gtt act gcc tac cta atg atg ttc tac acc ata ttc ggc ctc ttc 624

Met Val Thr Ala Tyr Leu Met Met Phe Tyr Thr Ile Phe Gly Leu Phe 195 200 205

Pagina 14

SEQLTXT aat gac atg agg aag acc ggg tca aat ttt tgg ttg agc gcc tct aca 672

Asn Asp Met Arg Lys Thr Gly Ser Asn Phe Trp Leu Ser Ala Ser Thr 210 215 220 gtg gtc aat tct gca tca tca ctt ttc tta gca ttg tat ttc acc caa 720

Val Val Asn Ser Ala Ser Ser Leu Phe Leu Ala Leu Tyr Phe Thr Gln 225 230 235 240 tgt att cta ggc aaa gaa gtt tcc gca tta act ctt ttt gaa ggt ttg 768

Cys Ile Leu Gly Lys Glu Val Ser Ala Leu Thr Leu Phe Glu Gly Leu 245 250 255 cct ttc att gta gtt gtt gtt ggt ttc aag cac aaa atc aag att gcc 816

Pro Phe Ile Val Val Val Val Gly Phe Lys His Lys Ile Lys Ile Ala 260 265 270 cag tat gcc ctg gag aaa ttt gaa aga gtc ggt tta tct aaa agg att 864

Gln Tyr Ala Leu Glu Lys Phe Glu Arg Val Gly Leu Ser Lys Arg Ile 275 280 285 act acc gat gaa atc gtt ttt gaa tcc gtg agc gaa gag ggt ggt cgt 912

Thr Thr Asp Glu Ile Val Phe Glu Ser Val Ser Glu Glu Gly Gly Arg 290 295 300 ttg att caa gac cat ttg ctt tgt att ttt gcc ttt atc gga tgc tct 960

Leu Ile Gln Asp His Leu Leu Cys Ile Phe Ala Phe Ile Gly Cys Ser 305 310 315 320 atg tat gct cac caa ttg aag act ttg aca aac ttc tgc ata tta tca 1008

Met Tyr Ala His Gln Leu Lys Thr Leu Thr Asn Phe Cys Ile Leu Ser 325 330 335 gca ttt atc cta att ttt gaa ttg att tta act cct aca ttt tat tct 1056

Ala Phe Ile Leu Ile Phe Glu Leu Ile Leu Thr Pro Thr Phe Tyr Ser 340 345 350 gct atc tta gcg ctt aga ctg gaa atg aat gtt atc cac aga tct act 1104

Ala Ile Leu Ala Leu Arg Leu Glu Met Asn Val Ile His Arg Ser Thr 355 360 365 att atc aag caa aca tta gaa gaa gac ggt gtt gtt cca tct aca gca 1152

Ile Ile Lys Gln Thr Leu Glu Glu Asp Gly Val Val Pro Ser Thr Ala 370 375 380 aga atc att tct aaa gca gaa aag aaa tcc gta tct tct ttc tta aat 1200

Arg Ile Ile Ser Lys Ala Glu Lys Lys Ser Val Ser Ser Phe Leu Asn 385 390 395 400 ctc agt gtg gtt gtc att atc atg aaa ctc tct gtc ata cta ttg ttc 1248

Leu Ser Val Val Val Ile Ile Met Lys Leu Ser Val Ile Leu Leu Phe 405 410 415

Pagina 15

SEQLTXT gtc ttc atc aac ttt tat aac ttt ggt gca aat tgg gtc aat gat gcc 1296

Val Phe Ile Asn Phe Tyr Asn Phe Gly Ala Asn Trp Val Asn Asp Ala 420 425 430 ttc aat tca ttg tac ttc gat aag gaa cgt gtt tct cta cca gat ttt 1344

Phe Asn Ser Leu Tyr Phe Asp Lys Glu Arg Val Ser Leu Pro Asp Phe 435 440 445 att acc tcg aat gcc tct gaa aac ttt aaa gag caa gct att gtt agt 1392

Ile Thr Ser Asn Ala Ser Glu Asn Phe Lys Glu Gln Ala Ile Val Ser 450 455 460 gtc acc cca tta tta tat tac aaa ccc att aag tcc tac caa cgc att 14409

Val Thr Pro Leu Leu Tyr Tyr Lys Pro Ile Lys Ser Tyr Gln Arg Ile 465 470 475 480 gag gat atg gtt ctt cta ttg ctt cgt aat gtc agt gtt gcc att cgt 1488

Glu Asp Met Val Leu Leu Leu Leu Arg Asn Val Ser Val Ala Ile Arg 485 490 495 gat agg ttc gtc agt aaa tta gtt ctt tcc gcc tta gta tgc agt gct 1536

Asp Arg Phe Val Ser Lys Leu Val Leu Ser Ala Leu Val Cys Ser Ala 500 505 510 gtc atc aat gtg tat tta tta aat gct gct aga att cat acc agt tat 1584

Val Ile Asn Val Tyr Leu Leu Asn Ala Ala Arg Ile His Thr Ser Tyr 515 520 525 act gca gac caa ttg gtg aag act gaa gtc acc aag aag tct ttt act 1632

Thr Ala Asp Gln Leu Val Lys Thr Glu Val Thr Lys Lys Ser Phe Thr 530 535 540 gct cct gta caa aag gct tct aca cca gtt tta acc aat aaa aca gtc 1680

Ala Pro Val Gln Lys Ala Ser Thr Pro Val Leu Thr Asn Lys Thr Val 545 550 555 560 att tct gga tcg aaa gtc aaa agt tta tca tct gcg caa tcg agc tca 1728

Ile Ser Gly Ser Lys Val Lys Ser Leu Ser Ser Ala Gln Ser Ser Ser 565 570 575 tca gga cct tca tca tct agt gag gaa gat gat tcc cgc gat att gaa 1776

Ser Gly Pro Ser Ser Ser Ser Glu Glu Asp Asp Ser Arg Asp Ile Glu 580 585 590 agc ttg gat aag aaa ata cgt cct tta gaa gaa tta gaa gca tta tta 1824

Ser Leu Asp Lys Lys Ile Arg Pro Leu Glu Glu Leu Glu Ala Leu Leu 595 600 605 agt agt gga aat aca aaa caa ttg aag aac aaa gag gtc gct gcc ttg 1872

Ser Ser Gly Asn Thr Lys Gln Leu Lys Asn Lys Glu Val Ala Ala Leu 610 615 620

Pagina 16

SEQLTXT gtt att cac ggt aag tta cct ttg tac gct ttg gag aaa aaa tta ggt 1920

Val Ile His Gly Lys Leu Pro Leu Tyr Ala Leu Glu Lys Lys Leu Gly 625 630 635 640 gat act acg aga gcg gtt gcg gta cgt agg aaa gct ctt tca att ttg 1968

Asp Thr Thr Arg Ala Val Ala Val Arg Arg Lys Ala Leu Ser Ile Leu 645 650 655 gca gaa gct cct gta tta gca tct gat cgt tta cca tat aaa aat tat 2016

Ala Glu Ala Pro Val Leu Ala Ser Asp Arg Leu Pro Tyr Lys Asn Tyr 660 665 670 gac tac gac cgc gta ttt ggc gct tgt tgt gaa aat gtt ata ggt tac 2064

Asp Tyr Asp Arg Val Phe Gly Ala Cys Cys Glu Asn Val Ile Gly Tyr 675 680 685 atg cct ttg ccc gtt ggt gtt ata ggc ccc ttg gtt atc gat ggt aca 2112

Met Pro Leu Pro Val Gly Val Ile Gly Pro Leu Val Ile Asp Gly Thr 690 695 700 tct tat cat ata cca atg gca act aca gag ggt tgt ttg gta gct tct 2160

Ser Tyr His Ile Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala Ser 705 710 715 720 gcc atg cgt ggc tgt aag gca atc aat gct ggc ggt ggt gca aca act 2208

Ala Met Arg Gly Cys Lys Ala Ile Asn Ala Gly Gly Gly Ala Thr Thr 725 730 735 gtt tta act aag gat ggt atg aca aga ggc cca gta gtc cgt ttc cca 2256

Val Leu Thr Lys Asp Gly Met Thr Arg Gly Pro Val Val Arg Phe Pro 740 745 750 act ttg aaa aga tct ggt gcc tgt aag ata tgg tta gac tca gaa gag 2304

Thr Leu Lys Arg Ser Gly Ala Cys Lys Ile Trp Leu Asp Ser Glu Glu 755 760 765 gga caa aac gca att aaa aaa gct ttt aac tct aca tca aga ttt gca 2352

Gly Gln Asn Ala Ile Lys Lys Ala Phe Asn Ser Thr Ser Arg Phe Ala 770 775 780 cgt ctg caa cat att caa act tgt cta gca gga gat tta ctc ttc atg 2400

Arg Leu Gln His Ile Gln Thr Cys Leu Ala Gly Asp Leu Leu Phe Met 785 790 795 800 aga ttt aga aca act act ggt gac gca atg ggt atg aat atg att tct 2448

Arg Phe Arg Thr Thr Thr Gly Asp Ala Met Gly Met Asn Met Ile Ser 805 810 815 aag ggt gtc gaa tac tca tta aag caa atg gta gaa gag tat ggc tgg 2496

Lys Gly Val Glu Tyr Ser Leu Lys Gln Met Val Glu Glu Tyr Gly Trp 820 825 830

Pagina 17

SEQLTXT gaa gat atg gag gtt gtc tcc gtt tct ggt aac tac tgt acc gac aaa 2544

Glu Asp Met Glu Val Val Ser Val Ser Gly Asn Tyr Cys Thr Asp Lys 835 840 845 aaa cca gct gcc atc aac tgg atc gaa ggt cgt ggt aag agt gtc gtc 2592

Lys Pro Ala Ala Ile Asn Trp Ile Glu Gly Arg Gly Lys Ser Val Val 850 855 860 gca gaa gct act att cct ggt gat gtt gtc aga aaa gtg tta aaa agt 2640

Ala Glu Ala Thr Ile Pro Gly Asp Val Val Arg Lys Val Leu Lys Ser 865 870 875 880 gat gtt tcc gca ttg gtt gag ttg aac att gct aag aat ttg gtt gga 2688

Asp Val Ser Ala Leu Val Glu Leu Asn Ile Ala Lys Asn Leu Val Gly 885 890 895 tct gca atg gct ggg tct gtt ggt gga ttt aac gca cat gca gct aat 2736

Ser Ala Met Ala Gly Ser Val Gly Gly Phe Asn Ala His Ala Ala Asn 900 905 910 tta gtg aca gct gtt ttc ttg gca tta gga caa gat cct gca caa aat 2784

Leu Val Thr Ala Val Phe Leu Ala Leu Gly Gln Asp Pro Ala Gln Asn 915 920 925 gtc gaa agt tcc aac tgt ata aca ttg atg aaa gaa gtg gac ggt gat 2832

Val Glu Ser Ser Asn Cys Ile Thr Leu Met Lys Glu Val Asp Gly Asp 930 935 940 ttg aga att tcc gta tcc atg cca tcc atc gaa gta ggt acc atc ggt 2880

Leu Arg Ile Ser Val Ser Met Pro Ser Ile Glu Val Gly Thr Ile Gly 945 950 955 960 ggt ggt act gtt cta gaa cca caa ggt gcc atg ttg gac tta tta ggt 2928

Gly Gly Thr Val Leu Glu Pro Gln Gly Ala Met Leu Asp Leu Leu Gly 965 970 975 gta aga ggc cca cat gct acc gct cct ggt acc aac gca cgt caa tta 2976

Val Arg Gly Pro His Ala Thr Ala Pro Gly Thr Asn Ala Arg Gln Leu 980 985 990 gca aga ata gtt gcc tgt gcc gtc ttg gca ggt gaa tta tcc tta tgt 3024

Ala Arg Ile Val Ala Cys Ala Val Leu Ala Gly Glu Leu Ser Leu Cys 995 1000 1005 gct gcc cta gca gcc ggc cat ttg gtt caa agt cat atg acc cac 3069

Ala Ala Leu Ala Ala Gly His Leu Val Gln Ser His Met Thr His 1010 1015 1020 aac agg aaa cct gct gaa cca aca aaa cct aac aat ttg gac gcc 3114

Asn Arg Lys Pro Ala Glu Pro Thr Lys Pro Asn Asn Leu Asp Ala 1025 1030 1035

Pagina 18

SEQLTXT act gat ata aat cgt ttg aaa gat ggg tcc gtc acc tgc att aaa 3159

Thr Asp Ile Asn Arg Leu Lys Asp Gly Ser Val Thr Cys Ile Lys 1040 1045 1050 tcc taa 3165

Ser <210> 7 <211> 1054 <212> PRT <213> Saccharomyces cerevisiae <400> 7

Met Pro Leu Leu Phe Lys Gly Leu Lys Gln Met Ala Lys Pro Ile Ala 1 5 10 15

Tyr Val Ser Arg Phe Ser Ala Lys Arg Pro Ile His Ile Ile Leu Phe

Ser Leu Ile Ile Ser Ala Phe Ala Tyr Leu Ser Val Ile Gln Tyr Tyr

Phe Asn Gly Trp Gln Leu Asp Ser Asn Ser Val Phe Glu Thr Ala Pro 60

Asn Lys Asp Ser Asn Thr Leu Phe Gln Glu Cys Ser His Tyr Tyr Arg 65 70 75 80

Asp Ser Ser Leu Asp Gly Trp Val Ser Ile Thr Ala His Glu Ala Ser 85 90 95

Glu Leu Pro Ala Pro His His Tyr Tyr Leu Leu Asn Leu Asn Phe Asn 100 105 119

Ser Pro Asn Glu Thr Asp Ser Ile Pro Glu Leu Ala Asn Thr Val Phe 115 120 125

Glu Lys Asp Asn Thr Lys Tyr Ile Leu Gln Glu Asp Leu Ser Val Ser 130 135 140

Pagina 19

SEQLTXT

Lys Glu Ile Ser Ser Thr Asp Gly Thr Lys Trp Arg Leu Arg Ser Asp 145 150 155 160

Arg Lys Ser Leu Phe Asp Val Lys Thr Leu Ala Tyr Ser Leu Tyr Asp 165 170 175

Val Phe Ser Glu Asn Val Thr Gln Ala Asp Pro Phe Asp Val Leu Ile 180 185 190

Met Val Thr Ala Tyr Leu Met Met Phe Tyr Thr Ile Phe Gly Leu Phe 195 200 205

Asn Asp Met Arg Lys Thr Gly Ser Asn Phe Trp Leu Ser Ala Ser Thr 210 215 220

Val Val Asn Ser Ala Ser Ser Leu Phe Leu Ala Leu Tyr Phe Thr Gln 225 230 235 240

Cys Ile Leu Gly Lys Glu Val Ser Ala Leu Thr Leu Phe Glu Gly Leu 245 250 255

Pro Phe Ile Val Val Val Val Gly Phe Lys His Lys Ile Lys Ile Ala 260 265 270

Gln Tyr Ala Leu Glu Lys Phe Glu Arg Val Gly Leu Ser Lys Arg Ile 275 280 285

Thr Thr Asp Glu Ile Val Phe Glu Ser Val Ser Glu Glu Gly Gly Arg 290 295 300

Leu Ile Gln Asp His Leu Leu Cys Ile Phe Ala Phe Ile Gly Cys Ser 305 310 315 320

Met Tyr Ala His Gln Leu Lys Thr Leu Thr Asn Phe Cys Ile Leu Ser 325 330 335

Ala Phe Ile Leu Ile Phe Glu Leu Ile Leu Thr Pro Thr Phe Tyr Ser 340 345 350

Pagina 20

SEQLTXT

Ala Ile Leu Ala Leu Arg Leu Glu Met Asn Val Ile His Arg Ser Thr 355 360 365

Ile Ile Lys Gln Thr Leu Glu Glu Asp Gly Val Val Pro Ser Thr Ala 370 375 380

Arg Ile Ile Ser Lys Ala Glu Lys Lys Ser Val Ser Ser Phe Leu Asn 385 390 395 400

Leu Ser Val Val Val Ile Ile Met Lys Leu Ser Val Ile Leu Leu Phe 405 410 415

Val Phe Ile Asn Phe Tyr Asn Phe Gly Ala Asn Trp Val Asn Asp Ala 420 425 430

Phe Asn Ser Leu Tyr Phe Asp Lys Glu Arg Val Ser Leu Pro Asp Phe 435 440 445

Ile Thr Ser Asn Ala Ser Glu Asn Phe Lys Glu Gln Ala Ile Val Ser 450 455 460

Val Thr Pro Leu Leu Tyr Tyr Lys Pro Ile Lys Ser Tyr Gln Arg Ile 465 470 475 480

Glu Asp Met Val Leu Leu Leu Leu Arg Asn Val Ser Val Ala Ile Arg 485 490 495

Asp Arg Phe Val Ser Lys Leu Val Leu Ser Ala Leu Val Cys Ser Ala 500 505 510

Val Ile Asn Val Tyr Leu Leu Asn Ala Ala Arg Ile His Thr Ser Tyr 515 520 525

Thr Ala Asp Gln Leu Val Lys Thr Glu Val Thr Lys Lys Ser Phe Thr 530 535 540

Ala Pro Val Gln Lys Ala Ser Thr Pro Val Leu Thr Asn Lys Thr Val 545 550 555 560

Pagina 21

SEQLTXT

Ile Ser Gly Ser Lys Val Lys Ser Leu Ser Ser Ala Gln Ser Ser Ser 565 570 575

Ser Gly Pro Ser Ser Ser Ser Glu Glu Asp Asp Ser Arg Asp Ile Glu 580 585 590

Ser Leu Asp Lys Lys Ile Arg Pro Leu Glu Glu Leu Glu Ala Leu Leu 595 600 605

Ser Ser Gly Asn Thr Lys Gln Leu Lys Asn Lys Glu Val Ala Ala Leu 610 615 620

Val Ile His Gly Lys Leu Pro Leu Tyr Ala Leu Glu Lys Lys Leu Gly 625 630 635 640

Asp Thr Thr Arg Ala Val Ala Val Arg Arg Lys Ala Leu Ser Ile Leu 645 650 655

Ala Glu Ala Pro Val Leu Ala Ser Asp Arg Leu Pro Tyr Lys Asn Tyr 660 665 670

Asp Tyr Asp Arg Val Phe Gly Ala Cys Cys Glu Asn Val Ile Gly Tyr 675 680 685

Met Pro Leu Pro Val Gly Val Ile Gly Pro Leu Val Ile Asp Gly Thr 690 695 700

Ser Tyr His Ile Pro Met Ala Thr Thr Glu Gly Cys Leu Val Ala Ser 705 710 715 720

Ala Met Arg Gly Cys Lys Ala Ile Asn Ala Gly Gly Gly Ala Thr Thr 725 730 735

Val Leu Thr Lys Asp Gly Met Thr Arg Gly Pro Val Val Arg Phe Pro 740 745 750

Thr Leu Lys Arg Ser Gly Ala Cys Lys Ile Trp Leu Asp Ser Glu Glu 755 760 765

Pagina 22

SEQLTXT

Gly Gln Asn Ala Ile Lys Lys Ala Phe Asn Ser Thr Ser Arg Phe Ala 770 775 780

Arg Leu Gln His Ile Gln Thr Cys Leu Ala Gly Asp Leu Leu Phe Met 785 790 795 800

Arg Phe Arg Thr Thr Thr Gly Asp Ala Met Gly Met Asn Met Ile Ser 805 810 815

Lys Gly Val Glu Tyr Ser Leu Lys Gln Met Val Glu Glu Tyr Gly Trp 820 825 830

Glu Asp Met Glu Val Val Ser Val Ser Gly Asn Tyr Cys Thr Asp Lys 835 840 845

Lys Pro Ala Ala Ile Asn Trp Ile Glu Gly Arg Gly Lys Ser Val Val 850 855 860

Ala Glu Ala Thr Ile Pro Gly Asp Val Val Arg Lys Val Leu Lys Ser 865 870 875 880

Asp Val Ser Ala Leu Val Glu Leu Asn Ile Ala Lys Asn Leu Val Gly 885 890 895

Ser Ala Met Ala Gly Ser Val Gly Gly Phe Asn Ala His Ala Ala Asn 900 905 910

Leu Val Thr Ala Val Phe Leu Ala Leu Gly Gln Asp Pro Ala Gln Asn 915 920 925

Val Glu Ser Ser Asn Cys Ile Thr Leu Met Lys Glu Val Asp Gly Asp 930 935 940

Leu Arg Ile Ser Val Ser Met Pro Ser Ile Glu Val Gly Thr Ile Gly 945 950 955 960

Gly Gly Thr Val Leu Glu Pro Gln Gly Ala Met Leu Asp Leu Leu Gly 965 970 975

Pagina 23

SEQLTXT

Val Arg Gly Pro His Ala Thr Ala Pro Gly Thr Asn Ala Arg Gln Leu 980 985 990

Ala Arg Ile Val Ala Cys Ala Val Leu Ala Gly Glu Leu Ser Leu Cys 995 1000 1005

Ala Ala Leu Ala Ala Gly His Leu Val Gln Ser His Met Thr His 1010 1015 1020

Asn Arg Lys Pro Ala Glu Pro Thr Lys Pro Asn Asn Leu Asp Ala 1025 1030 1035

Thr Asp Ile Asn Arg Leu Lys Asp Gly Ser Val Thr Cys Ile Lys 1040 1045 1050

Ser <2105 8 <211> 502 <212> PRT <213> Artificial Sequence <220> <223> Truncated HMGR <400> 8

Met Val Leu Thr Asn Lys Thr Val Ile Ser Gly Ser Lys Val Lys Ser 1 5 10 15

Leu Ser Ser Ala Gln Ser Ser Ser Ser Gly Pro Ser Ser Ser Ser Glu

Glu Asp Asp Ser Arg Asp Ile Glu Ser Leu Asp Lys Lys Ile Arg Pro

Leu Glu Glu Leu Glu Ala Leu Leu Ser Ser Gly Asn Thr Lys Gln Leu 60

Lys Asn Lys Glu Val Ala Ala Leu Val Ile His Gly Lys Leu Pro Leu 65 70 75 80

Pagina 24

SEQLTXT

Tyr Ala Leu Glu Lys Lys Leu Gly Asp Thr Thr Arg Ala Val Ala Val 85 90 95

Arg Arg Lys Ala Leu Ser Ile Leu Ala Glu Ala Pro Val Leu Ala Ser 100 105 110

Asp Arg Leu Pro Tyr Lys Asn Tyr Asp Tyr Asp Arg Val Phe Gly Ala 115 120 125

Cys Cys Glu Asn Val Ile Gly Tyr Met Pro Leu Pro Val Gly Val Ile 130 135 140

Gly Pro Leu Val Ile Asp Gly Thr Ser Tyr His Ile Pro Met Ala Thr 145 150 155 160

Thr Glu Gly Cys Leu Val Ala Ser Ala Met Arg Gly Cys Lys Ala Ile 165 170 175

Asn Ala Gly Gly Gly Ala Thr Thr Val Leu Thr Lys Asp Gly Met Thr 180 185 190

Arg Gly Pro Val Val Arg Phe Pro Thr Leu Lys Arg Ser Gly Ala Cys 195 200 205

Lys Ile Trp Leu Asp Ser Glu Glu Gly Gln Asn Ala Ile Lys Lys Ala 210 215 220

Phe Asn Ser Thr Ser Arg Phe Ala Arg Leu Gln His Ile Gln Thr Cys 225 230 235 240

Leu Ala Gly Asp Leu Leu Phe Met Arg Phe Arg Thr Thr Thr Gly Asp 245 250 255

Ala Met Gly Met Asn Met Ile Ser Lys Gly Val Glu Tyr Ser Leu Lys 260 265 270

Gln Met Val Glu Glu Tyr Gly Trp Glu Asp Met Glu Val Val Ser Val 275 280 285

Pagina 25

SEQLTXT

Ser Gly Asn Tyr Cys Thr Asp Lys Lys Pro Ala Ala Ile Asn Trp Ile 290 295 300

Glu Gly Arg Gly Lys Ser Val Val Ala Glu Ala Thr Ile Pro Gly Asp 305 310 315 320

Val Val Arg Lys Val Leu Lys Ser Asp Val Ser Ala Leu Val Glu Leu 325 330 335

Asn Ile Ala Lys Asn Leu Val Gly Ser Ala Met Ala Gly Ser Val Gly 340 345 350

Gly Phe Asn Ala His Ala Ala Asn Leu Val Thr Ala Val Phe Leu Ala 355 360 365

Leu Gly Gln Asp Pro Ala Gln Asn Val Glu Ser Ser Asn Cys Ile Thr 370 375 380

Leu Met Lys Glu Val Asp Gly Asp Leu Arg Ile Ser Val Ser Met Pro 385 390 395 400

Ser Ile Glu Val Gly Thr Ile Gly Gly Gly Thr Val Leu Glu Pro Gln 405 410 415

Gly Ala Met Leu Asp Leu Leu Gly Val Arg Gly Pro His Ala Thr Ala 420 425 430

Pro Gly Thr Asn Ala Arg Gln Leu Ala Arg Ile Val Ala Cys Ala Val 435 440 445

Leu Ala Gly Glu Leu Ser Leu Cys Ala Ala Leu Ala Ala Gly His Leu 450 455 460

Val Gln Ser His Met Thr His Asn Arg Lys Pro Ala Glu Pro Thr Lys 465 470 475 480

Pro Asn Asn Leu Asp Ala Thr Asp Ile Asn Arg Leu Lys Asp Gly Ser 485 490 495

Pagina 26

SEQLTXT

Val Thr Cys Ile Lys Ser 500 <210> 9 <211> 1332 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(1332) <400> 9 atg tca tta ccg ttc tta act tct gca ccg gga aag gtt att att ttt 48

Met Ser Leu Pro Phe Leu Thr Ser Ala Pro Gly Lys Val Ile Ile Phe 1 5 10 15 ggt gaa cac tct gct gtg tac aac aag cct gcc gtc get get agt gtg 96

Gly Glu His Ser Ala Val Tyr Asn Lys Pro Ala Val Ala Ala Ser Val tct gcg ttg aga acc tac ctg cta ata agc gag tca tct gca cca gat 144

Ser Ala Leu Arg Thr Tyr Leu Leu Ile Ser Glu Ser Ser Ala Pro Asp act att gaa ttg gac ttc ccg gac att agc ttt aat cat aag tgg tcc 192

Thr Ile Glu Leu Asp Phe Pro Asp Ile Ser Phe Asn His Lys Trp Ser 60 atc aat gat ttc aat gcc atc acc gag gat caa gta aat tct caa aaa 240

Ile Asn Asp Phe Asn Ala Ile Thr Glu Asp Gln Val Asn Ser Gln Lys 65 70 75 80 ttg gcc aag gct caa caa gcc acc gat ggc ttg tct cag gaa ctc gtt 288

Leu Ala Lys Ala Gln Gln Ala Thr Asp Gly Leu Ser Gln Glu Leu Val 85 90 95 agt ctt ttg gat ccg ttg tta gct caa cta tcc gaa tcc ttc cac tac 336

Ser Leu Leu Asp Pro Leu Leu Ala Gln Leu Ser Glu Ser Phe His Tyr 100 105 110 cat gca gcg ttt tgt ttc ctg tat atg ttt gtt tgc cta tgc ccc cat 384

His Ala Ala Phe Cys Phe Leu Tyr Met Phe Val Cys Leu Cys Pro His 115 120 125 gcc aag aat att aag ttt tct tta aag tct act tta ccc atc ggt gct 432

Ala Lys Asn Ile Lys Phe Ser Leu Lys Ser Thr Leu Pro Ile Gly Ala 130 135 140

Pagina 27

SEQLTXT ggg ttg ggc tca agc gcc tct att tct gta tca ctg gcc tta get atg 480

Gly Leu Gly Ser Ser Ala Ser Ile Ser Val Ser Leu Ala Leu Ala Met 145 150 155 160 gcc tac ttg ggg ggg tta ata gga tct aat gac ttg gaa aag ctg tca 528

Ala Tyr Leu Gly Gly Leu Ile Gly Ser Asn Asp Leu Glu Lys Leu Ser 165 170 175 gaa aac gat aag cat ata gtg aat caa tgg gcc ttc ata ggt gaa aag 576

Glu Asn Asp Lys His Ile Val Asn Gln Trp Ala Phe Ile Gly Glu Lys 180 185 190 tgt att cac ggt acc cct tca gga ata gat aac gct gtg gcc act tat 624

Cys Ile His Gly Thr Pro Ser Gly Ile Asp Asn Ala Val Ala Thr Tyr 195 200 205 ggt aat gcc ctg cta ttt gaa aaa gac tca cat aat gga aca ata aac 672

Gly Asn Ala Leu Leu Phe Glu Lys Asp Ser His Asn Gly Thr Ile Asn 210 215 220 aca aac aat ttt aag ttc tta gat gat ttc cca gcc att cca atg atc 720

Thr Asn Asn Phe Lys Phe Leu Asp Asp Phe Pro Ala Ile Pro Met Ile 225 230 235 240 cta acc tat act aga att cca agg tct aca aaa gat ctt gtt gct cgc 768

Leu Thr Tyr Thr Arg Ile Pro Arg Ser Thr Lys Asp Leu Val Ala Arg 245 250 255 gtt cgt gtg ttg gtc acc gag aaa ttt cct gaa gtt atg aag cca gtt 816

Val Arg Val Leu Val Thr Glu Lys Phe Pro Glu Val Met Lys Pro Val 260 265 270 cta gat gcc atg ggt gaa tgt gcc cta caa ggc tta gag atc atg act 864

Leu Asp Ala Met Gly Glu Cys Ala Leu Gln Gly Leu Glu Ile Met Thr 275 280 285 aag tta agt aaa tgt aaa ggc acc gat gac gag gct gta gaa act aat 912

Lys Leu Ser Lys Cys Lys Gly Thr Asp Asp Glu Ala Val Glu Thr Asn 290 295 300 aat gaa ctg tat gaa caa cta ttg gaa ttg ata aga ata aat cat gga 960

Asn Glu Leu Tyr Glu Gln Leu Leu Glu Leu Ile Arg Ile Asn His Gly 305 310 315 320 ctg ctt gtc tca atc ggt gtt tct cat cct gga tta gaa ctt att aaa 1008

Leu Leu Val Ser Ile Gly Val Ser His Pro Gly Leu Glu Leu Ile Lys 325 330 335 aat ctg agc gat gat ttg aga att ggc tcc aca aaa ctt acc ggt gct 1056

Asn Leu Ser Asp Asp Leu Arg Ile Gly Ser Thr Lys Leu Thr Gly Ala 340 345 350

Pagina 28

SEQLTXT ggt ggc ggc ggt tgc tct ttg act ttg tta cga aga gac att act caa 1104

Gly Gly Gly Gly Cys Ser Leu Thr Leu Leu Arg Arg Asp Ile Thr Gln 355 360 365 gag caa att gac agt ttc aaa aag aaa ttg caa gat gat ttt agt tac 1152

Glu Gln Ile Asp Ser Phe Lys Lys Lys Leu Gln Asp Asp Phe Ser Tyr 370 375 380 gag aca ttt gaa aca gac ttg ggt ggg act ggc tgc tgt ttg tta agc 1200

Glu Thr Phe Glu Thr Asp Leu Gly Gly Thr Gly Cys Cys Leu Leu Ser 385 390 395 400 gca aaa aat ttg aat aaa gat ctt aaa atc aaa tcc cta gta ttc caa 1248

Ala Lys Asn Leu Asn Lys Asp Leu Lys Ile Lys Ser Leu Val Phe Gln 405 410 415 tta ttt gaa aat aaa act acc aca aag caa caa att gac gat cta tta 1296

Leu Phe Glu Asn Lys Thr Thr Thr Lys Gln Gln Ile Asp Asp Leu Leu 420 425 430 ttg cca gga aat acg aat tta cca tgg act tca taa 1332

Leu Pro Gly Asn Thr Asn Leu Pro Trp Thr Ser 435 440 <210> 10 <211> 443 <212> PRT <213> Saccharomyces cerevisiae <400> 10

Met Ser Leu Pro Phe Leu Thr Ser Ala Pro Gly Lys Val Ile Ile Phe 1 5 10 15

Gly Glu His Ser Ala Val Tyr Asn Lys Pro Ala Val Ala Ala Ser Val

Ser Ala Leu Arg Thr Tyr Leu Leu Ile Ser Glu Ser Ser Ala Pro Asp

Thr Ile Glu Leu Asp Phe Pro Asp Ile Ser Phe Asn His Lys Trp Ser 60

Ile Asn Asp Phe Asn Ala Ile Thr Glu Asp Gln Val Asn Ser Gln Lys 65 70 75 80

Pagina 29

SEQLTXT

Leu Ala Lys Ala Gln Gln Ala Thr Asp Gly Leu Ser Gln Glu Leu Val 85 90 95

Ser Leu Leu Asp Pro Leu Leu Ala Gln Leu Ser Glu Ser Phe His Tyr 100 105 110

His Ala Ala Phe Cys Phe Leu Tyr Met Phe Val Cys Leu Cys Pro His 115 120 125

Ala Lys Asn Ile Lys Phe Ser Leu Lys Ser Thr Leu Pro Ile Gly Ala 130 135 140

Gly Leu Gly Ser Ser Ala Ser Ile Ser Val Ser Leu Ala Leu Ala Met 145 150 155 160

Ala Tyr Leu Gly Gly Leu Ile Gly Ser Asn Asp Leu Glu Lys Leu Ser 165 170 175

Glu Asn Asp Lys His Ile Val Asn Gln Trp Ala Phe Ile Gly Glu Lys 180 185 190

Cys Ile His Gly Thr Pro Ser Gly Ile Asp Asn Ala Val Ala Thr Tyr 195 200 205

Gly Asn Ala Leu Leu Phe Glu Lys Asp Ser His Asn Gly Thr Ile Asn 210 215 220

Thr Asn Asn Phe Lys Phe Leu Asp Asp Phe Pro Ala Ile Pro Met Ile 225 230 235 240

Leu Thr Tyr Thr Arg Ile Pro Arg Ser Thr Lys Asp Leu Val Ala Arg 245 250 255

Val Arg Val Leu Val Thr Glu Lys Phe Pro Glu Val Met Lys Pro Val 260 265 270

Leu Asp Ala Met Gly Glu Cys Ala Leu Gln Gly Leu Glu Ile Met Thr 275 280 285

Pagina 30

SEQLTXT

Lys Leu Ser Lys Cys Lys Gly Thr Asp Asp Glu Ala Val Glu Thr Asn 290 295 300

Asn Glu Leu Tyr Glu Gln Leu Leu Glu Leu Ile Arg Ile Asn His Gly 305 310 315 320

Leu Leu Val Ser Ile Gly Val Ser His Pro Gly Leu Glu Leu Ile Lys 325 330 335

Asn Leu Ser Asp Asp Leu Arg Ile Gly Ser Thr Lys Leu Thr Gly Ala 340 345 350

Gly Gly Gly Gly Cys Ser Leu Thr Leu Leu Arg Arg Asp Ile Thr Gln 355 360 365

Glu Gln Ile Asp Ser Phe Lys Lys Lys Leu Gln Asp Asp Phe Ser Tyr 370 375 380

Glu Thr Phe Glu Thr Asp Leu Gly Gly Thr Gly Cys Cys Leu Leu Ser 385 390 395 400

Ala Lys Asn Leu Asn Lys Asp Leu Lys Ile Lys Ser Leu Val Phe Gln 405 410 415

Leu Phe Glu Asn Lys Thr Thr Thr Lys Gln Gln Ile Asp Asp Leu Leu 420 425 430

Leu Pro Gly Asn Thr Asn Leu Pro Trp Thr Ser 435 440 <210> 11 <211> 1356 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(1356) <400> 11

Pagina 31

SEQLTXT atg tca gag ttg aga gcc ttc agt gcc cca ggg aaa gcg tta cta gct 48

Met Ser Glu Leu Arg Ala Phe Ser Ala Pro Gly Lys Ala Leu Leu Ala 1 5 10 15 ggt gga tat tta gtt tta gat aca aaa tat gaa gca ttt gta gtc gga 96

Gly Gly Tyr Leu Val Leu Asp Thr Lys Tyr Glu Ala Phe Val Val Gly tta tcg gca aga atg cat gct gta gcc cat cct tac ggt tca ttg caa 144

Leu Ser Ala Arg Met His Ala Val Ala His Pro Tyr Gly Ser Leu Gln gag tct gat aag ttt gaa gtg cgt gtg aaa agt aaa caa ttt aaa gat 192

Glu Ser Asp Lys Phe Glu Val Arg Val Lys Ser Lys Gln Phe Lys Asp 60 ggg gag tgg ctg tac cat ata agt cct aaa act ggc ttc att cct gtt 240

Gly Glu Trp Leu Tyr His Ile Ser Pro Lys Thr Gly Phe Ile Pro Val 65 70 75 80 tcg ata ggc gga tct aag aac cct ttc att gaa aaa gtt atc gct aac 288

Ser Ile Gly Gly Ser Lys Asn Pro Phe Ile Glu Lys Val Ile Ala Asn 85 90 95 gta ttt agc tac ttt aaa cct aac atg gac gac tac tgc aat aga aac 336

Val Phe Ser Tyr Phe Lys Pro Asn Met Asp Asp Tyr Cys Asn Arg Asn 100 105 110 ttg ttc gtt att gat att ttc tct gat gat gcc tac cat tct cag gag 384

Leu Phe Val Ile Asp Ile Phe Ser Asp Asp Ala Tyr His Ser Gln Glu 115 120 125 gat agc gtt acc gaa cat cgt ggc aac aga aga ttg agt ttt cat tcg 432

Asp Ser Val Thr Glu His Arg Gly Asn Arg Arg Leu Ser Phe His Ser 130 135 140 cac aga att gaa gaa gtt ccc aaa aca ggg ctg ggc tcc tcg gca ggt 480

His Arg Ile Glu Glu Val Pro Lys Thr Gly Leu Gly Ser Ser Ala Gly 145 150 155 160 tta gtc aca gtt tta act aca gct ttg gcc tcc ttt ttt gta tcg gac 528

Leu Val Thr Val Leu Thr Thr Ala Leu Ala Ser Phe Phe Val Ser Asp 165 170 175 ctg gaa aat aat gta gac aaa tat aga gaa gtt att cat aat tta tca 576

Leu Glu Asn Asn Val Asp Lys Tyr Arg Glu Val Ile His Asn Leu Ser 180 185 190 caa gtt gct cat tgt caa gct cag ggt aaa att gga agc ggg ttt gat 624

Gln Val Ala His Cys Gln Ala Gln Gly Lys Ile Gly Ser Gly Phe Asp 195 200 205

Pagina 32

SEQLTXT gta gcg gcg gca gca tat gga tct atc aga tat aga aga ttc cca ccc 672

Val Ala Ala Ala Ala Tyr Gly Ser Ile Arg Tyr Arg Arg Phe Pro Pro 210 215 220 gca tta atc tct aat ttg cca gat att gga agt gct act tac ggc agt 720

Ala Leu Ile Ser Asn Leu Pro Asp Ile Gly Ser Ala Thr Tyr Gly Ser 225 230 235 240 aaa ctg gcg cat ttg gtt gat gaa gaa gac tgg aat att acg att aaa 768

Lys Leu Ala His Leu Val Asp Glu Glu Asp Trp Asn Ile Thr Ile Lys 245 250 255 agt aac cat tta cct tcg gga tta act tta tgg atg ggc gat att aag 816

Ser Asn His Leu Pro Ser Gly Leu Thr Leu Trp Met Gly Asp Ile Lys 260 265 270 aat ggt tca gaa aca gta aaa ctg gtc cag aag gta aaa aat tgg tat 864

Asn Gly Ser Glu Thr Val Lys Leu Val Gln Lys Val Lys Asn Trp Tyr 275 280 285 gat tcg cat atg cca gaa agc ttg aaa ata tat aca gaa ctc gat cat 912

Asp Ser His Met Pro Glu Ser Leu Lys Ile Tyr Thr Glu Leu Asp His 290 295 300 gca aat tct aga ttt atg gat gga cta tct aaa cta gat cgc tta cac 960

Ala Asn Ser Arg Phe Met Asp Gly Leu Ser Lys Leu Asp Arg Leu His 305 310 315 320 gag act cat gac gat tac agc gat cag ata ttt gag tct ctt gag agg 1008

Glu Thr His Asp Asp Tyr Ser Asp Gln Ile Phe Glu Ser Leu Glu Arg 325 330 335 aat gac tgt acc tgt caa aag tat cct gaa atc aca gaa gtt aga gat 1056

Asn Asp Cys Thr Cys Gln Lys Tyr Pro Glu Ile Thr Glu Val Arg Asp 340 345 350 gca gtt gcc aca att aga cgt tcc ttt aga aaa ata act aaa gaa tct 1104

Ala Val Ala Thr Ile Arg Arg Ser Phe Arg Lys Ile Thr Lys Glu Ser 355 360 365 ggt gcc gat atc gaa cct ccc gta caa act agc tta ttg gat gat tgc 1152

Gly Ala Asp Ile Glu Pro Pro Val Gln Thr Ser Leu Leu Asp Asp Cys 370 375 380 cag acc tta aaa gga gtt ctt act tgc tta ata cct ggt gct ggt ggt 1200

Gln Thr Leu Lys Gly Val Leu Thr Cys Leu Ile Pro Gly Ala Gly Gly 385 390 395 400 tat gac gcc att gca gtg att act aag caa gat gtt gat ctt agg gct 1248

Tyr Asp Ala Ile Ala Val Ile Thr Lys Gln Asp Val Asp Leu Arg Ala 405 410 415

Pagina 33

SEQLTXT caa acc gct aat gac aaa aga ttt tct aag gtt caa tgg ctg gat gta 1296

Gln Thr Ala Asn Asp Lys Arg Phe Ser Lys Val Gln Trp Leu Asp Val 420 425 430 act cag gct gac tgg ggt gtt agg aaa gaa aaa gat ccg gaa act tat 1344

Thr Gln Ala Asp Trp Gly Val Arg Lys Glu Lys Asp Pro Glu Thr Tyr 435 440 445 ctt gat aaa taa 1356

Leu Asp Lys 450 <21e> 12 <211> 451 <212> PRT <213> Saccharomyces cerevisiae <400> 12

Met Ser Glu Leu Arg Ala Phe Ser Ala Pro Gly Lys Ala Leu Leu Ala 1 5 10 15

Gly Gly Tyr Leu Val Leu Asp Thr Lys Tyr Glu Ala Phe Val Val Gly

Leu Ser Ala Arg Met His Ala Val Ala His Pro Tyr Gly Ser Leu Gln

Glu Ser Asp Lys Phe Glu Val Arg Val Lys Ser Lys Gln Phe Lys Asp 60

Gly Glu Trp Leu Tyr His Ile Ser Pro Lys Thr Gly Phe Ile Pro Val 65 70 75 80

Ser Ile Gly Gly Ser Lys Asn Pro Phe Ile Glu Lys Val Ile Ala Asn 85 90 95

Val Phe Ser Tyr Phe Lys Pro Asn Met Asp Asp Tyr Cys Asn Arg Asn 100 105 119

Leu Phe Val Ile Asp Ile Phe Ser Asp Asp Ala Tyr His Ser Gln Glu 115 120 125

Pagina 34

SEQLTXT

Asp Ser Val Thr Glu His Arg Gly Asn Arg Arg Leu Ser Phe His Ser 130 135 140

His Arg Ile Glu Glu Val Pro Lys Thr Gly Leu Gly Ser Ser Ala Gly 145 150 155 160

Leu Val Thr Val Leu Thr Thr Ala Leu Ala Ser Phe Phe Val Ser Asp 165 170 175

Leu Glu Asn Asn Val Asp Lys Tyr Arg Glu Val Ile His Asn Leu Ser 180 185 190

Gln Val Ala His Cys Gln Ala Gln Gly Lys Ile Gly Ser Gly Phe Asp 195 200 205

Val Ala Ala Ala Ala Tyr Gly Ser Ile Arg Tyr Arg Arg Phe Pro Pro 210 215 220

Ala Leu Ile Ser Asn Leu Pro Asp Ile Gly Ser Ala Thr Tyr Gly Ser 225 230 235 240

Lys Leu Ala His Leu Val Asp Glu Glu Asp Trp Asn Ile Thr Ile Lys 245 250 255

Ser Asn His Leu Pro Ser Gly Leu Thr Leu Trp Met Gly Asp Ile Lys 260 265 270

Asn Gly Ser Glu Thr Val Lys Leu Val Gln Lys Val Lys Asn Trp Tyr 275 280 285

Asp Ser His Met Pro Glu Ser Leu Lys Ile Tyr Thr Glu Leu Asp His 290 295 300

Ala Asn Ser Arg Phe Met Asp Gly Leu Ser Lys Leu Asp Arg Leu His 305 310 315 320

Glu Thr His Asp Asp Tyr Ser Asp Gln Ile Phe Glu Ser Leu Glu Arg 325 330 335

Pagina 35

SEQLTXT

Asn Asp Cys Thr Cys Gln Lys Tyr Pro Glu Ile Thr Glu Val Arg Asp 340 345 350

Ala Val Ala Thr Ile Arg Arg Ser Phe Arg Lys Ile Thr Lys Glu Ser 355 360 365

Gly Ala Asp Ile Glu Pro Pro Val Gln Thr Ser Leu Leu Asp Asp Cys 370 375 380

Gln Thr Leu Lys Gly Val Leu Thr Cys Leu Ile Pro Gly Ala Gly Gly 385 390 395 400

Tyr Asp Ala Ile Ala Val Ile Thr Lys Gln Asp Val Asp Leu Arg Ala 405 410 415

Gln Thr Ala Asn Asp Lys Arg Phe Ser Lys Val Gln Trp Leu Asp Val 420 425 430

Thr Gln Ala Asp Trp Gly Val Arg Lys Glu Lys Asp Pro Glu Thr Tyr 435 440 445

Leu Asp Lys 450 <21e> 13 <211> 1191 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(1191) <400> 13 atg acc gtt tac aca gca tcc gtt acc gca ccc gtc aac atc gca acc 48

Met Thr Val Tyr Thr Ala Ser Val Thr Ala Pro Val Asn Ile Ala Thr 1 5 10 15 ctt aag tat tgg ggg aaa agg gac acg aag ttg aat ctg ccc acc aat 96

Leu Lys Tyr Trp Gly Lys Arg Asp Thr Lys Leu Asn Leu Pro Thr Asn tcg tcc ata tca gtg act tta tcg caa gat gac ctc aga acg tta acc 144

Pagina 36

SEQLTXT

Ser Ser Ile Ser Val Thr Leu Ser Gln Asp Asp Leu Arg Thr Leu Thr tct gcg gct act gca cct gag ttt gaa cgc gac act ttg tgg tta aat 192

Ser Ala Ala Thr Ala Pro Glu Phe Glu Arg Asp Thr Leu Trp Leu Asn 60 gga gaa cca cac agc atc gac aat gaa aga act caa aat tgt ctg cgc 240

Gly Glu Pro His Ser Ile Asp Asn Glu Arg Thr Gln Asn Cys Leu Arg 65 70 75 80 gac cta cgc caa tta aga aag gaa atg gaa tcg aag gac gcc tca ttg 288

Asp Leu Arg Gln Leu Arg Lys Glu Met Glu Ser Lys Asp Ala Ser Leu 85 90 95 ccc aca tta tct caa tgg aaa ctt cac att gtc tcc gaa aat aac ttt 336

Pro Thr Leu Ser Gln Trp Lys Leu His Ile Val Ser Glu Asn Asn Phe 100 105 110 cct aca gca gct ggt tta gct tcc tcc get get gge ttt get gca ttg 384

Pro Thr Ala Ala Gly Leu Ala Ser Ser Ala Ala Gly Phe Ala Ala Leu 115 120 125 gtc tct gca att gct aag tta tac caa tta cca cag tca act tca gaa 432

Val Ser Ala Ile Ala Lys Leu Tyr Gln Leu Pro Gln Ser Thr Ser Glu 130 135 140 ata tct aga ata gca aga aag ggg tct ggt tca gct tgt aga tcg ttg 480

Ile Ser Arg Ile Ala Arg Lys Gly Ser Gly Ser Ala Cys Arg Ser Leu 145 150 155 160 ttt ggc gga tac gtg gcc tgg gaa atg ggc aaa gct gaa gat ggt cat 528

Phe Gly Gly Tyr Val Ala Trp Glu Met Gly Lys Ala Glu Asp Gly His 165 170 175 gat tcc atg gca gta caa atc gca gac agc tct gac tgg cct cag atg 576

Asp Ser Met Ala Val Gln Ile Ala Asp Ser Ser Asp Trp Pro Gln Met 180 185 190 aaa gct tgt gtc cta gtt gtc agc gat att aaa aag gat gtg agt tcc 624

Lys Ala Cys Val Leu Val Val Ser Asp Ile Lys Lys Asp Val Ser Ser 195 200 205 act cag ggt atg caa ttg acc gtg gca acc tcc gaa cta ttt aaa gaa 672

Thr Gln Gly Met Gln Leu Thr Val Ala Thr Ser Glu Leu Phe Lys Glu 210 215 220 aga att gaa cat gtc gta cca aag aga ttt gaa gtc atg cgt aaa gcc 720

Arg Ile Glu His Val Val Pro Lys Arg Phe Glu Val Met Arg Lys Ala 225 230 235 240 att gtt gaa aaa gat ttc gcc acc ttt gca aag gaa aca atg atg gat 768

Pagina 37

SEQLTXT

Ile Val Glu Lys Asp Phe Ala Thr Phe Ala Lys Glu Thr Met Met Asp 245 250 255 tcc aac tct ttc cat gcc aca tgt ttg gac tct ttc cct cca ata ttc 816

Ser Asn Ser Phe His Ala Thr Cys Leu Asp Ser Phe Pro Pro Ile Phe 260 265 270 tac atg aac gac act tcc aag cgt atc atc agt tgg tgc cac acc att 864

Tyr Met Asn Asp Thr Ser Lys Arg Ile Ile Ser Trp Cys His Thr Ile 275 280 285 aat cag ttt tac gga gaa aca atc gtt gca tac acg ttt gat gca ggt 912

Asn Gln Phe Tyr Gly Glu Thr Ile Val Ala Tyr Thr Phe Asp Ala Gly 290 295 300 cca aat gct gtg ttg tac tac tta gct gaa aat gag tcg aaa atc ttt 960

Pro Asn Ala Val Leu Tyr Tyr Leu Ala Glu Asn Glu Ser Lys Ile Phe 305 310 315 320 gca ttt atc tat aaa ttg ttt ggc tct gtt cct gga tgg gac aag aaa 1008

Ala Phe Ile Tyr Lys Leu Phe Gly Ser Val Pro Gly Trp Asp Lys Lys 325 330 335 ttt act gct gag cag ctt gag gct ttc aac cat caa ttt gaa tct tct 1056

Phe Thr Ala Glu Gln Leu Glu Ala Phe Asn His Gln Phe Glu Ser Ser 340 345 350 aac ttt act gca cgt gaa ttg gat ctt gag ttg caa aag ggt gtt gcc 1104

Asn Phe Thr Ala Arg Glu Leu Asp Leu Glu Leu Gln Lys Gly Val Ala 355 360 365 aga gtg att tta act caa gtc ggt tca ggc cca caa gaa aca aat gaa 1152

Arg Val Ile Leu Thr Gln Val Gly Ser Gly Pro Gln Glu Thr Asn Glu 370 375 380 tct ttg att gac gca aag act ggt cta cca aag aaa taa 1191

Ser Leu Ile Asp Ala Lys Thr Gly Leu Pro Lys Lys 385 390 395 <210> 14 <211> 396 <212> PRT <213> Saccharomyces cerevisiae <400> 14

Met Thr Val Tyr Thr Ala Ser Val Thr Ala Pro Val Asn Ile Ala Thr 1 5 10 15

Leu Lys Tyr Trp Gly Lys Arg Asp Thr Lys Leu Asn Leu Pro Thr Asn

Pagina 38

SEQLTXT

Ser Ser Ile Ser Val Thr Leu Ser Gln Asp Asp Leu Arg Thr Leu Thr

Ser Ala Ala Thr Ala Pro Glu Phe Glu Arg Asp Thr Leu Trp Leu Asn 60

Gly Glu Pro His Ser Ile Asp Asn Glu Arg Thr Gln Asn Cys Leu Arg 65 70 75 80

Asp Leu Arg Gln Leu Arg Lys Glu Met Glu Ser Lys Asp Ala Ser Leu 85 90 95

Pro Thr Leu Ser Gln Trp Lys Leu His Ile Val Ser Glu Asn Asn Phe 100 105 110

Pro Thr Ala Ala Gly Leu Ala Ser Ser Ala Ala Gly Phe Ala Ala Leu 115 120 125

Val Ser Ala Ile Ala Lys Leu Tyr Gln Leu Pro Gln Ser Thr Ser Glu 130 135 140

Ile Ser Arg Ile Ala Arg Lys Gly Ser Gly Ser Ala Cys Arg Ser Leu 145 150 155 160

Phe Gly Gly Tyr Val Ala Trp Glu Met Gly Lys Ala Glu Asp Gly His 165 170 175

Asp Ser Met Ala Val Gln Ile Ala Asp Ser Ser Asp Trp Pro Gln Met 180 185 190

Lys Ala Cys Val Leu Val Val Ser Asp Ile Lys Lys Asp Val Ser Ser 195 200 205

Thr Gln Gly Met Gln Leu Thr Val Ala Thr Ser Glu Leu Phe Lys Glu 210 215 220

Arg Ile Glu His Val Val Pro Lys Arg Phe Glu Val Met Arg Lys Ala

Pagina 39

SEQLTXT

225 230 235 240

Ile Val Glu Lys Asp Phe Ala Thr Phe Ala Lys Glu Thr Met Met Asp 245 250 255

Ser Asn Ser Phe His Ala Thr Cys Leu Asp Ser Phe Pro Pro Ile Phe 260 265 270

Tyr Met Asn Asp Thr Ser Lys Arg Ile Ile Ser Trp Cys His Thr Ile 275 280 285

Asn Gln Phe Tyr Gly Glu Thr Ile Val Ala Tyr Thr Phe Asp Ala Gly 290 295 300

Pro Asn Ala Val Leu Tyr Tyr Leu Ala Glu Asn Glu Ser Lys Ile Phe 305 310 315 320

Ala Phe Ile Tyr Lys Leu Phe Gly Ser Val Pro Gly Trp Asp Lys Lys 325 330 335

Phe Thr Ala Glu Gln Leu Glu Ala Phe Asn His Gln Phe Glu Ser Ser 340 345 350

Asn Phe Thr Ala Arg Glu Leu Asp Leu Glu Leu Gln Lys Gly Val Ala 355 360 365

Arg Val Ile Leu Thr Gln Val Gly Ser Gly Pro Gln Glu Thr Asn Glu 370 375 380

Ser Leu Ile Asp Ala Lys Thr Gly Leu Pro Lys Lys 385 390 395 <210> 15 <211> 867 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(867)

Pagina 40

SEQLTXT

<400> 15 atg act gcc gac aac aat agt atg ccc cat ggt gca gta tct agt tac 48

Met Thr Ala Asp Asn Asn Ser Met Pro His Gly Ala Val Ser Ser Tyr 1 5 10 15 gcc aaa tta gtg caa aac caa aca cct gaa gac att ttg gaa gag ttt 96

Ala Lys Leu Val Gln Asn Gln Thr Pro Glu Asp Ile Leu Glu Glu Phe cct gaa att att cca tta caa caa aga cct aat acc cga tct agt gag 144

Pro Glu Ile Ile Pro Leu Gln Gln Arg Pro Asn Thr Arg Ser Ser Glu acg tca aat gac gaa agc gga gaa aca tgt ttt tct ggt cat gat gag 192

Thr Ser Asn Asp Glu Ser Gly Glu Thr Cys Phe Ser Gly His Asp Glu 60 gag caa att aag tta atg aat gaa aat tgt att gtt ttg gat tgg gac 240

Glu Gln Ile Lys Leu Met Asn Glu Asn Cys Ile Val Leu Asp Trp Asp 65 70 75 80 gat aat gct att ggt gcc ggt acc aag aaa gtt tgt cat tta atg gaa 288

Asp Asn Ala Ile Gly Ala Gly Thr Lys Lys Val Cys His Leu Met Glu 85 90 95 aat att gaa aag ggt tta cta cat cgt gca ttc tcc gtc ttt att ttc 336

Asn Ile Glu Lys Gly Leu Leu His Arg Ala Phe Ser Val Phe Ile Phe 100 105 110 aat gaa caa ggt gaa tta ctt tta caa caa aga gcc act gaa aaa ata 384

Asn Glu Gln Gly Glu Leu Leu Leu Gln Gln Arg Ala Thr Glu Lys Ile 115 120 125 act ttc cct gat ctt tgg act aac aca tgc tgc tct cat cca cta tgt 432

Thr Phe Pro Asp Leu Trp Thr Asn Thr Cys Cys Ser His Pro Leu Cys 130 135 140 att gat gac gaa tta ggt ttg aag ggt aag cta gac gat aag att aag 480

Ile Asp Asp Glu Leu Gly Leu Lys Gly Lys Leu Asp Asp Lys Ile Lys 145 150 155 160 ggc gct att act gcg gcg gtg aga aaa cta gat cat gaa tta ggt att 528

Gly Ala Ile Thr Ala Ala Val Arg Lys Leu Asp His Glu Leu Gly Ile 165 170 175 cca gaa gat gaa act aag aca agg ggt aag ttt cac ttt tta aac aga 576

Pro Glu Asp Glu Thr Lys Thr Arg Gly Lys Phe His Phe Leu Asn Arg 180 185 190 atc cat tac atg gca cca agc aat gaa cca tgg ggt gaa cat gaa att 624

Ile His Tyr Met Ala Pro Ser Asn Glu Pro Trp Gly Glu His Glu Ile

Pagina 41

SEQLTXT

195 200 205 gat tac atc cta ttt tat aag atc aac gct aaa gaa aac ttg act gtc 672

Asp Tyr Ile Leu Phe Tyr Lys Ile Asn Ala Lys Glu Asn Leu Thr Val 210 215 220 aac cca aac gtc aat gaa gtt aga gac ttc aaa tgg gtt tca cca aat 720

Asn Pro Asn Val Asn Glu Val Arg Asp Phe Lys Trp Val Ser Pro Asn 225 230 235 240 gat ttg aaa act atg ttt gct gac cca agt tac aag ttt acg cct tgg 768

Asp Leu Lys Thr Met Phe Ala Asp Pro Ser Tyr Lys Phe Thr Pro Trp 245 250 255 ttt aag att att tgc gag aat tac tta ttc aac tgg tgg gag caa tta 816

Phe Lys Ile Ile Cys Glu Asn Tyr Leu Phe Asn Trp Trp Glu Gln Leu 260 265 270 gat gac ctt tct gaa gtg gaa aat gac agg caa att cat aga atg cta 864

Asp Asp Leu Ser Glu Val Glu Asn Asp Arg Gln Ile His Arg Met Leu 275 280 285 taa 867 <210> 16 <211> 288 <212> PRT <213> Saccharomyces cerevisiae <400> 16

Met Thr Ala Asp Asn Asn Ser Met Pro His Gly Ala Val Ser Ser Tyr 1 5 10 15

Ala Lys Leu Val Gln Asn Gln Thr Pro Glu Asp Ile Leu Glu Glu Phe

Pro Glu Ile Ile Pro Leu Gln Gln Arg Pro Asn Thr Arg Ser Ser Glu

Thr Ser Asn Asp Glu Ser Gly Glu Thr Cys Phe Ser Gly His Asp Glu 60

Glu Gln Ile Lys Leu Met Asn Glu Asn Cys Ile Val Leu Asp Trp Asp 65 70 75 80

Pagina 42

SEQLTXT

Asp Asn Ala Ile Gly Ala Gly Thr Lys Lys Val Cys His Leu Met Glu 85 90 95

Asn Ile Glu Lys Gly Leu Leu His Arg Ala Phe Ser Val Phe Ile Phe 100 105 110

Asn Glu Gln Gly Glu Leu Leu Leu Gln Gln Arg Ala Thr Glu Lys Ile 115 120 125

Thr Phe Pro Asp Leu Trp Thr Asn Thr Cys Cys Ser His Pro Leu Cys 130 135 140

Ile Asp Asp Glu Leu Gly Leu Lys Gly Lys Leu Asp Asp Lys Ile Lys 145 150 155 160

Gly Ala Ile Thr Ala Ala Val Arg Lys Leu Asp His Glu Leu Gly Ile 165 170 175

Pro Glu Asp Glu Thr Lys Thr Arg Gly Lys Phe His Phe Leu Asn Arg 180 185 190

Ile His Tyr Met Ala Pro Ser Asn Glu Pro Trp Gly Glu His Glu Ile 195 200 205

Asp Tyr Ile Leu Phe Tyr Lys Ile Asn Ala Lys Glu Asn Leu Thr Val 210 215 220

Asn Pro Asn Val Asn Glu Val Arg Asp Phe Lys Trp Val Ser Pro Asn 225 230 235 240

Asp Leu Lys Thr Met Phe Ala Asp Pro Ser Tyr Lys Phe Thr Pro Trp 245 250 255

Phe Lys Ile Ile Cys Glu Asn Tyr Leu Phe Asn Trp Trp Glu Gln Leu 260 265 270

Asp Asp Leu Ser Glu Val Glu Asn Asp Arg Gln Ile His Arg Met Leu 275 280 285

Pagina 43

SEQLTXT

<210> 17 <211> 1059 <212> DNA <213> Saccharomyces cerevisiae <220> <221> CDS <222> (1)..(1959) <400> 17 atg gct tca gaa aaa gaa att agg aga gag aga ttc ttg aac gtt ttc 48

Met Ala Ser Glu Lys Glu Ile Arg Arg Glu Arg Phe Leu Asn Val Phe 1 5 10 15 cct aaa tta gta gag gaa ttg aac gca tcg ctt ttg gct tac ggt atg 96

Pro Lys Leu Val Glu Glu Leu Asn Ala Ser Leu Leu Ala Tyr Gly Met cct aag gaa gca tgt gac tgg tat gcc cac tca ttg aac tac aac act 144

Pro Lys Glu Ala Cys Asp Trp Tyr Ala His Ser Leu Asn Tyr Asn Thr cca ggc ggt aag cta aat aga ggt ttg tcc gtt gtg gac acg tat gct 192

Pro Gly Gly Lys Leu Asn Arg Gly Leu Ser Val Val Asp Thr Tyr Ala 60 att ctc tcc aac aag acc gtt gaa caa ttg ggg caa gaa gaa tac gaa 240

Ile Leu Ser Asn Lys Thr Val Glu Gln Leu Gly Gln Glu Glu Tyr Glu 65 70 75 80 aag gtt gcc att cta ggt tgg tgc att gag ttg ttg cag gct tac ttc 288

Lys Val Ala Ile Leu Gly Trp Cys Ile Glu Leu Leu Gln Ala Tyr Phe 85 90 95 ttg gtc gcc gat gat atg atg gac aag tcc att acc aga aga ggc caa 336

Leu Val Ala Asp Asp Met Met Asp Lys Ser Ile Thr Arg Arg Gly Gln 100 105 110 cca tgt tgg tac aag gtt cct gaa gtt ggg gaa att gcc atc aat gac 384

Pro Cys Trp Tyr Lys Val Pro Glu Val Gly Glu Ile Ala Ile Asn Asp 115 120 125 gca ttc atg tta gag gct gct atc tac aag ctt ttg aaa tct cac ttc 432

Ala Phe Met Leu Glu Ala Ala Ile Tyr Lys Leu Leu Lys Ser His Phe 130 135 140 aga aac gaa aaa tac tac ata gat atc acc gaa ttg ttc cat gag gtc 480

Arg Asn Glu Lys Tyr Tyr Ile Asp Ile Thr Glu Leu Phe His Glu Val 145 150 155 160 acc ttc caa acc gaa ttg ggc caa ttg atg gac tta atc act gca cct 528

Pagina 44

SEQLTXT

Thr Phe Gln Thr Glu Leu Gly Gln Leu Met Asp Leu Ile Thr Ala Pro 165 170 175 gaa gac aaa gtc gac ttg agt aag ttc tcc cta aag aag cac tcc ttc 576

Glu Asp Lys Val Asp Leu Ser Lys Phe Ser Leu Lys Lys His Ser Phe 180 185 190 ata gtt act ttc aag act gct tac tat tct ttc tac ttg cct gtc gca 624

Ile Val Thr Phe Lys Thr Ala Tyr Tyr Ser Phe Tyr Leu Pro Val Ala 195 200 205 ttg gcc atg tac gtt gcc ggt atc acg gat gaa aag gat ttg aaa caa 672

Leu Ala Met Tyr Val Ala Gly Ile Thr Asp Glu Lys Asp Leu Lys Gln 210 215 220 gcc aga gat gtc ttg att cca ttg ggt gaa tac ttc caa att caa gat 720

Ala Arg Asp Val Leu Ile Pro Leu Gly Glu Tyr Phe Gln Ile Gln Asp 225 230 235 240 gac tac tta gac tgc ttc ggt acc cca gaa cag atc ggt aag atc ggt 768

Asp Tyr Leu Asp Cys Phe Gly Thr Pro Glu Gln Ile Gly Lys Ile Gly 245 250 255 aca gat atc caa gat aac aaa tgt tct tgg gta atc aac aag gca ttg 816

Thr Asp Ile Gln Asp Asn Lys Cys Ser Trp Val Ile Asn Lys Ala Leu 260 265 270 gaa ctt gct tcc gca gaa caa aga aag act tta gac gaa aat tac ggt 864

Glu Leu Ala Ser Ala Glu Gln Arg Lys Thr Leu Asp Glu Asn Tyr Gly 275 280 285 aag aag gac tca gtc gca gaa gcc aaa tgc aaa aag att ttc aat gac 912

Lys Lys Asp Ser Val Ala Glu Ala Lys Cys Lys Lys Ile Phe Asn Asp 290 295 300 ttg aaa att gaa cag cta tac cac gaa tat gaa gag tct att gcc aag 960

Leu Lys Ile Glu Gln Leu Tyr His Glu Tyr Glu Glu Ser Ile Ala Lys 305 310 315 320 gat ttg aag gcc aaa att tct cag gtc gat gag tct cgt ggc ttc aaa 1008

Asp Leu Lys Ala Lys Ile Ser Gln Val Asp Glu Ser Arg Gly Phe Lys 325 330 335 gct gat gtc tta act gcg ttc ttg aac aaa gtt tac aag aga agc aaa 1056

Ala Asp Val Leu Thr Ala Phe Leu Asn Lys Val Tyr Lys Arg Ser Lys 340 345 350 tag 1059 <210> 18 <211> 352

Pagina 45

SEQLTXT

<212> PRT <213> Saccharomyces cerevisiae <400> 18

Met Ala Ser Glu Lys Glu Ile Arg Arg Glu Arg Phe Leu Asn Val Phe 1 5 10 15

Pro Lys Leu Val Glu Glu Leu Asn Ala Ser Leu Leu Ala Tyr Gly Met

Pro Lys Glu Ala Cys Asp Trp Tyr Ala His Ser Leu Asn Tyr Asn Thr

Pro Gly Gly Lys Leu Asn Arg Gly Leu Ser Val Val Asp Thr Tyr Ala 60

Ile Leu Ser Asn Lys Thr Val Glu Gln Leu Gly Gln Glu Glu Tyr Glu 65 70 75 80

Lys Val Ala Ile Leu Gly Trp Cys Ile Glu Leu Leu Gln Ala Tyr Phe 85 90 95

Leu Val Ala Asp Asp Met Met Asp Lys Ser Ile Thr Arg Arg Gly Gln 100 105 110

Pro Cys Trp Tyr Lys Val Pro Glu Val Gly Glu Ile Ala Ile Asn Asp 115 120 125

Ala Phe Met Leu Glu Ala Ala Ile Tyr Lys Leu Leu Lys Ser His Phe 130 135 140

Arg Asn Glu Lys Tyr Tyr Ile Asp Ile Thr Glu Leu Phe His Glu Val 145 150 155 160

Thr Phe Gln Thr Glu Leu Gly Gln Leu Met Asp Leu Ile Thr Ala Pro 165 170 175

Glu Asp Lys Val Asp Leu Ser Lys Phe Ser Leu Lys Lys His Ser Phe 180 185 190

Pagina 46

SEQLTXT

Ile Val Thr Phe Lys Thr Ala Tyr Tyr Ser Phe Tyr Leu Pro Val Ala 195 200 205

Leu Ala Met Tyr Val Ala Gly Ile Thr Asp Glu Lys Asp Leu Lys Gln 210 215 220

Ala Arg Asp Val Leu Ile Pro Leu Gly Glu Tyr Phe Gln Ile Gln Asp 225 230 235 240

Asp Tyr Leu Asp Cys Phe Gly Thr Pro Glu Gln Ile Gly Lys Ile Gly 245 250 255

Thr Asp Ile Gln Asp Asn Lys Cys Ser Trp Val Ile Asn Lys Ala Leu 260 265 270

Glu Leu Ala Ser Ala Glu Gln Arg Lys Thr Leu Asp Glu Asn Tyr Gly 275 280 285

Lys Lys Asp Ser Val Ala Glu Ala Lys Cys Lys Lys Ile Phe Asn Asp 290 295 300

Leu Lys Ile Glu Gln Leu Tyr His Glu Tyr Glu Glu Ser Ile Ala Lys 305 310 315 320

Asp Leu Lys Ala Lys Ile Ser Gln Val Asp Glu Ser Arg Gly Phe Lys 325 330 335

Ala Asp Val Leu Thr Ala Phe Leu Asn Lys Val Tyr Lys Arg Ser Lys 340 345 350 <210> 19 <211> 1683 <212> DNA <213> Citrus junos <220> <221> CDS <222> (1)..(1683) <400> 19

Pagina 47

SEQLTXT atg aag gat atg tca ata ccg tta ttg gcc gcc gtc agt age tct acc 48

Met Lys Asp Met Ser Ile Pro Leu Leu Ala Ala Val Ser Ser Ser Thr 1 5 10 15 gaa gaa acg gtt cgt cca atc gct gat ttt cat cca acg cta tgg ggt 96

Glu Glu Thr Val Arg Pro Ile Ala Asp Phe His Pro Thr Leu Trp Gly aat cat ttt ttg aaa agc gct gct gac gta gag act ata gat gcc gcc 144

Asn His Phe Leu Lys Ser Ala Ala Asp Val Glu Thr Ile Asp Ala Ala aca cag gaa caa cac gcc gcc ttg aaa caa gaa gtc aga cgt atg ata 192

Thr Gln Glu Gln His Ala Ala Leu Lys Gln Glu Val Arg Arg Met Ile 60 aca act act gct aat aaa cta gca cag aaa tta cac atg ata gat gca 240

Thr Thr Thr Ala Asn Lys Leu Ala Gln Lys Leu His Met Ile Asp Ala 65 70 75 80 gtc caa aga tta ggt gtc gct tat cat ttt gaa aaa gaa att gaa gac 288

Val Gln Arg Leu Gly Val Ala Tyr His Phe Glu Lys Glu Ile Glu Asp 85 90 95 gaa ttg ggt aag gtg agt cat gat ttg gat tca gac gat tta tat gtg 336

Glu Leu Gly Lys Val Ser His Asp Leu Asp Ser Asp Asp Leu Tyr Val 100 105 110 gtg agt ttg aga ttc agg ctg ttt aga cag caa ggc gtt aaa att tcg 384

Val Ser Leu Arg Phe Arg Leu Phe Arg Gln Gln Gly Val Lys Ile Ser 115 120 125 tgc gat gtt ttc gac aag ttt aag gat gac gag ggt aaa ttc aaa gag 432

Cys Asp Val Phe Asp Lys Phe Lys Asp Asp Glu Gly Lys Phe Lys Glu 130 135 140 tca cta atc aat gat atc aga ggc atg tta tcc tta tat gaa gcc gct 480

Ser Leu Ile Asn Asp Ile Arg Gly Met Leu Ser Leu Tyr Glu Ala Ala 145 150 155 160 tat tta gca att agg ggg gaa gat ata ttg gac gaa gca ata gtg ttc 528

Tyr Leu Ala Ile Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Val Phe 165 170 175 aca aca aca cat ctg aaa tcc gta ata tca att tca gat cat agt cat 576

Thr Thr Thr His Leu Lys Ser Val Ile Ser Ile Ser Asp His Ser His 180 185 190 gcc aat tct aac ttg gct gaa caa ata cgt cac tct ttg caa att cca 624

Ala Asn Ser Asn Leu Ala Glu Gln Ile Arg His Ser Leu Gln Ile Pro 195 200 205

Pagina 48

SEQLTXT ttg aga aag gca gct gct aga ttg gaa gct aga tat ttc ctg gac att 672

Leu Arg Lys Ala Ala Ala Arg Leu Glu Ala Arg Tyr Phe Leu Asp Ile 210 215 220 tac agt aga gac gat ttg cac gat gag acc ctt ctg aaa ttt gct aag 720

Tyr Ser Arg Asp Asp Leu His Asp Glu Thr Leu Leu Lys Phe Ala Lys 225 230 235 240 cta gat ttc aat ata ctt caa gcc gca cat cag aag gaa gct agt atc 768

Leu Asp Phe Asn Ile Leu Gln Ala Ala His Gln Lys Glu Ala Ser Ile 245 250 255 atg act aga tgg tgg aat gac ttg ggt ttt cct aaa aaa gtt cct tac 816

Met Thr Arg Trp Trp Asn Asp Leu Gly Phe Pro Lys Lys Val Pro Tyr 260 265 270 gcc aga gat aga atc att gag aca tat ata tgg atg tta tta ggt gtt 864

Ala Arg Asp Arg Ile Ile Glu Thr Tyr Ile Trp Met Leu Leu Gly Val 275 280 285 tct tat gaa cca aat ttg gct ttc ggc agg att ttc gct tcc aaa gtc 912

Ser Tyr Glu Pro Asn Leu Ala Phe Gly Arg Ile Phe Ala Ser Lys Val 290 295 300 gta tgt atg att aca act att gat gat act ttc gac gct tat ggt aca 960

Val Cys Met Ile Thr Thr Ile Asp Asp Thr Phe Asp Ala Tyr Gly Thr 305 310 315 320 ttt gag gaa ctg act ttg ttt act gaa gca gtc aca aga tgg gat att 1008

Phe Glu Glu Leu Thr Leu Phe Thr Glu Ala Val Thr Arg Trp Asp Ile 325 330 335 ggc ttg ata gat act ctg cca gaa tac atg aag ttt att gta aaa gcg 1056

Gly Leu Ile Asp Thr Leu Pro Glu Tyr Met Lys Phe Ile Val Lys Ala 340 345 350 tta ttg gat ata tat aga gaa gca gaa gaa gaa ttg gca aaa gaa ggc 1104

Leu Leu Asp Ile Tyr Arg Glu Ala Glu Glu Glu Leu Ala Lys Glu Gly 355 360 365 cgt tcg tat ggc ata cct tac gct aag cag atg atg caa gag cta ata 1152

Arg Ser Tyr Gly Ile Pro Tyr Ala Lys Gln Met Met Gln Glu Leu Ile 370 375 380 att ctt tac ttc aca gaa gca aaa tgg cta tac aag ggt tac gta cca 1200

Ile Leu Tyr Phe Thr Glu Ala Lys Trp Leu Tyr Lys Gly Tyr Val Pro 385 390 395 400 acc ttc gac gaa tac aaa tct gtt gct cta aga tca atc ggc tta agg 1248

Thr Phe Asp Glu Tyr Lys Ser Val Ala Leu Arg Ser Ile Gly Leu Arg 405 410 415

Pagina 49

SEQLTXT acc ttg gcg gtt gct tct ttc gtt gac cta ggt gat ttt att gca aca 1296

Thr Leu Ala Val Ala Ser Phe Val Asp Leu Gly Asp Phe Ile Ala Thr 420 425 430 aaa gac aac ttc gaa tgc ata ttg aag aac gcg aag agt tta aaa gct 1344

Lys Asp Asn Phe Glu Cys Ile Leu Lys Asn Ala Lys Ser Leu Lys Ala 435 440 445 aca gag acc att gga aga tta atg gac gac atc gcg ggg tat aaa ttc 1392

Thr Glu Thr Ile Gly Arg Leu Met Asp Asp Ile Ala Gly Tyr Lys Phe 450 455 460 gaa caa aaa agg ggt cac aat ccg tca gca gtt gaa tgc tac aaa aat 14409

Glu Gln Lys Arg Gly His Asn Pro Ser Ala Val Glu Cys Tyr Lys Asn 465 470 475 480 caa cat gga gta tcc gaa gaa gag gct gtg aag gaa ctt tta cta gaa 1488

Gln His Gly Val Ser Glu Glu Glu Ala Val Lys Glu Leu Leu Leu Glu 485 490 495 gtt gca aac tca tgg aag gac atc aac gaa gaa ctg cta aat cca aca 1536

Val Ala Asn Ser Trp Lys Asp Ile Asn Glu Glu Leu Leu Asn Pro Thr 500 505 510 aca gta cca cta ccc atg cta cag aga ctt cta tac ttc gcg cgt tct 1584

Thr Val Pro Leu Pro Met Leu Gln Arg Leu Leu Tyr Phe Ala Arg Ser 515 520 525 ggt cac ttt att tat gat gac ggg cat gac agg tac act cat tca ctg 1632

Gly His Phe Ile Tyr Asp Asp Gly His Asp Arg Tyr Thr His Ser Leu 530 535 540 atg atg aaa cgt caa gtt gcg ctt cta cta aca gaa cct ctg gct att 1680

Met Met Lys Arg Gln Val Ala Leu Leu Leu Thr Glu Pro Leu Ala Ile 545 550 555 560 tga 1683 <210> 20 <211> 560 <212> PRT <213> Citrus junos <400> 20

Met Lys Asp Met Ser Ile Pro Leu Leu Ala Ala Val Ser Ser Ser Thr 1 5 10 15

Glu Glu Thr Val Arg Pro Ile Ala Asp Phe His Pro Thr Leu Trp Gly

Pagina 50

SEQLTXT

Asn His Phe Leu Lys Ser Ala Ala Asp Val Glu Thr Ile Asp Ala Ala

Thr Gln Glu Gln His Ala Ala Leu Lys Gln Glu Val Arg Arg Met Ile 60

Thr Thr Thr Ala Asn Lys Leu Ala Gln Lys Leu His Met Ile Asp Ala 65 70 75 80

Val Gln Arg Leu Gly Val Ala Tyr His Phe Glu Lys Glu Ile Glu Asp 85 90 95

Glu Leu Gly Lys Val Ser His Asp Leu Asp Ser Asp Asp Leu Tyr Val 100 105 110

Val Ser Leu Arg Phe Arg Leu Phe Arg Gln Gln Gly Val Lys Ile Ser 115 120 125

Cys Asp Val Phe Asp Lys Phe Lys Asp Asp Glu Gly Lys Phe Lys Glu 130 135 140

Ser Leu Ile Asn Asp Ile Arg Gly Met Leu Ser Leu Tyr Glu Ala Ala 145 150 155 160

Tyr Leu Ala Ile Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Val Phe 165 170 175

Thr Thr Thr His Leu Lys Ser Val Ile Ser Ile Ser Asp His Ser His 180 185 190

Ala Asn Ser Asn Leu Ala Glu Gln Ile Arg His Ser Leu Gln Ile Pro 195 200 205

Leu Arg Lys Ala Ala Ala Arg Leu Glu Ala Arg Tyr Phe Leu Asp Ile 210 215 220

Tyr Ser Arg Asp Asp Leu His Asp Glu Thr Leu Leu Lys Phe Ala Lys 225 230 235 240

Pagina 51

SEQLTXT

Leu Asp Phe Asn Ile Leu Gln Ala Ala His Gln Lys Glu Ala Ser Ile 245 250 255

Met Thr Arg Trp Trp Asn Asp Leu Gly Phe Pro Lys Lys Val Pro Tyr 260 265 270

Ala Arg Asp Arg Ile Ile Glu Thr Tyr Ile Trp Met Leu Leu Gly Val 275 280 285

Ser Tyr Glu Pro Asn Leu Ala Phe Gly Arg Ile Phe Ala Ser Lys Val 290 295 300

Val Cys Met Ile Thr Thr Ile Asp Asp Thr Phe Asp Ala Tyr Gly Thr 305 310 315 320

Phe Glu Glu Leu Thr Leu Phe Thr Glu Ala Val Thr Arg Trp Asp Ile 325 330 335

Gly Leu Ile Asp Thr Leu Pro Glu Tyr Met Lys Phe Ile Val Lys Ala 340 345 350

Leu Leu Asp Ile Tyr Arg Glu Ala Glu Glu Glu Leu Ala Lys Glu Gly 355 360 365

Arg Ser Tyr Gly Ile Pro Tyr Ala Lys Gln Met Met Gln Glu Leu Ile 370 375 380

Ile Leu Tyr Phe Thr Glu Ala Lys Trp Leu Tyr Lys Gly Tyr Val Pro 385 390 395 400

Thr Phe Asp Glu Tyr Lys Ser Val Ala Leu Arg Ser Ile Gly Leu Arg 405 410 415

Thr Leu Ala Val Ala Ser Phe Val Asp Leu Gly Asp Phe Ile Ala Thr 420 425 430

Lys Asp Asn Phe Glu Cys Ile Leu Lys Asn Ala Lys Ser Leu Lys Ala 435 440 445

Pagina 52

SEQLTXT

Thr Glu Thr Ile Gly Arg Leu Met Asp Asp Ile Ala Gly Tyr Lys Phe 450 455 460

Glu Gln Lys Arg Gly His Asn Pro Ser Ala Val Glu Cys Tyr Lys Asn 465 470 475 480

Gln His Gly Val Ser Glu Glu Glu Ala Val Lys Glu Leu Leu Leu Glu 485 490 495

Val Ala Asn Ser Trp Lys Asp Ile Asn Glu Glu Leu Leu Asn Pro Thr 500 505 510

Thr Val Pro Leu Pro Met Leu Gln Arg Leu Leu Tyr Phe Ala Arg Ser 515 520 525

Gly His Phe Ile Tyr Asp Asp Gly His Asp Arg Tyr Thr His Ser Leu 530 535 540

Met Met Lys Arg Gln Val Ala Leu Leu Leu Thr Glu Pro Leu Ala Ile 545 550 555 560 <21e> 21 <211> 1683 <212> DNA <213> Citrus junos <220> <221> CDS <222> (1)..(1683) <400> 21 atg aag gac atg tct att cct tta ttg gca gct gtt agt tca tct act 48

Met Lys Asp Met Ser Ile Pro Leu Leu Ala Ala Val Ser Ser Ser Thr 1 5 10 15 gaa gaa acc gtc aga cca att gct gac ttc cac cct acc tta tgg ggt 96

Glu Glu Thr Val Arg Pro Ile Ala Asp Phe His Pro Thr Leu Trp Gly aac cac ttt ttg aag agt gcc gct gat gtt gaa aca att gac gct gct 144

Asn His Phe Leu Lys Ser Ala Ala Asp Val Glu Thr Ile Asp Ala Ala

Pagina 53

SEQLTXT act caa gaa caa cac gca gct tta aag caa gaa gtc aga cgt atg atc 192

Thr Gln Glu Gln His Ala Ala Leu Lys Gln Glu Val Arg Arg Met Ile 60 act acc acc gcc aat aag ttg gct caa aag tta cat atg att gat gcc 240

Thr Thr Thr Ala Asn Lys Leu Ala Gln Lys Leu His Met Ile Asp Ala 65 70 75 80 gtt caa aga tta ggt gtc gct tat cat ttc gaa aag gag att gaa gat 288

Val Gln Arg Leu Gly Val Ala Tyr His Phe Glu Lys Glu Ile Glu Asp 85 90 95 gaa ttg ggt aag gtt tct cac gac cta gac tct gac gat tta tac gtc 336

Glu Leu Gly Lys Val Ser His Asp Leu Asp Ser Asp Asp Leu Tyr Val 100 105 110 gtc tct tta aga ttt aga ttg ttc aga caa caa ggt gtt aaa att agt 384

Val Ser Leu Arg Phe Arg Leu Phe Arg Gln Gln Gly Val Lys Ile Ser 115 120 125 tgt gat gtc ttt gat aag ttc aag gac gat gag ggt aag ttc aag gaa 432

Cys Asp Val Phe Asp Lys Phe Lys Asp Asp Glu Gly Lys Phe Lys Glu 130 135 140 tct ttg att aat gac atc aga ggc atg tta tcc ttg tac gaa gcc gcc 480

Ser Leu Ile Asn Asp Ile Arg Gly Met Leu Ser Leu Tyr Glu Ala Ala 145 150 155 160 tac cta gcc att aga ggt gaa gat atc tta gat gaa gct att gtt ttc 528

Tyr Leu Ala Ile Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Val Phe 165 170 175 act act act cac tta aaa tct gtc att agt att tct gac cac tct cat 576

Thr Thr Thr His Leu Lys Ser Val Ile Ser Ile Ser Asp His Ser His 180 185 190 gct aat tct aac tta gct gaa caa atc cgt cat tct cta caa att cca 624

Ala Asn Ser Asn Leu Ala Glu Gln Ile Arg His Ser Leu Gln Ile Pro 195 200 205 tta aga aag gct gca gct aga ttg gaa gct aga tat ttc cta gac atc 672

Leu Arg Lys Ala Ala Ala Arg Leu Glu Ala Arg Tyr Phe Leu Asp Ile 210 215 220 tac tcc aga gat gat tta cac gat gaa act ttg ttg aag ttc gct aaa 720

Tyr Ser Arg Asp Asp Leu His Asp Glu Thr Leu Leu Lys Phe Ala Lys 225 230 235 240 ttg gat ttt aac att ttg caa gct gct cac caa aag gag gca tcc att 768

Leu Asp Phe Asn Ile Leu Gln Ala Ala His Gln Lys Glu Ala Ser Ile 245 250 255

Pagina 54

SEQLTXT atg act aga tgg tgg aac gat ttg ggt ttc cca aaa aag gtt cca tat 816

Met Thr Arg Trp Trp Asn Asp Leu Gly Phe Pro Lys Lys Val Pro Tyr 260 265 270 gct aga gat aga att atc gaa acc tac atc tgg atg ttg ttg ggt gtc 864

Ala Arg Asp Arg Ile Ile Glu Thr Tyr Ile Trp Met Leu Leu Gly Val 275 280 285 agt tat gaa cca aac ttg gcc ttt ggt aga att ttc gct tca aag gtt 912

Ser Tyr Glu Pro Asn Leu Ala Phe Gly Arg Ile Phe Ala Ser Lys Val 290 295 300 gtc tgt atg att act acc att gac gat act ttt gat gcc tac ggt act 960

Val Cys Met Ile Thr Thr Ile Asp Asp Thr Phe Asp Ala Tyr Gly Thr 305 310 315 320 ttc gaa gaa ttg acc tta ttc acc gaa gct gtt acc aga tgg gat att 1008

Phe Glu Glu Leu Thr Leu Phe Thr Glu Ala Val Thr Arg Trp Asp Ile 325 330 335 ggc cta att gac acc ttg cca gaa tac atg aaa ttt atc gtt aaa gct 1056

Gly Leu Ile Asp Thr Leu Pro Glu Tyr Met Lys Phe Ile Val Lys Ala 340 345 350 ttg ttg gac att tac aga gaa gcc gaa gaa gaa ttg gca aag gaa ggt 1104

Leu Leu Asp Ile Tyr Arg Glu Ala Glu Glu Glu Leu Ala Lys Glu Gly 355 360 365 cgt agt tat ggt atc cca tac gct aaa caa atg atg caa gaa ttg att 1152

Arg Ser Tyr Gly Ile Pro Tyr Ala Lys Gln Met Met Gln Glu Leu Ile 370 375 380 att ttg tac ttc act gaa gct aag tgg ttg tac aaa ggt tac gtt cca 1200

Ile Leu Tyr Phe Thr Glu Ala Lys Trp Leu Tyr Lys Gly Tyr Val Pro 385 390 395 400 acc ttc gat gaa tat aag tcc gtc gcc tta aga tct att ggt tta aga 1248

Thr Phe Asp Glu Tyr Lys Ser Val Ala Leu Arg Ser Ile Gly Leu Arg 405 410 415 act cta gct gtt gct tcc ttc gtc gat ttg ggt gat ttt att gcc act 1296

Thr Leu Ala Val Ala Ser Phe Val Asp Leu Gly Asp Phe Ile Ala Thr 420 425 430 aag gac aat ttc gaa tgc atc ttg aaa aac gct aag tcc cta aag gca 1344

Lys Asp Asn Phe Glu Cys Ile Leu Lys Asn Ala Lys Ser Leu Lys Ala 435 440 445 act gaa act atc ggt aga tta atg gac gat att gct ggt tac aaa ttt 1392

Thr Glu Thr Ile Gly Arg Leu Met Asp Asp Ile Ala Gly Tyr Lys Phe 450 455 460

Pagina 55

SEQLTXT gaa caa aag aga ggt cat aat cca tcc gca gtt gaa tgt tac aaa aat 14409

Glu Gln Lys Arg Gly His Asn Pro Ser Ala Val Glu Cys Tyr Lys Asn 465 470 475 480 caa cat ggt gtc tcc gaa gaa gaa gct gtc aag gaa tta ttg ttg gaa 1488

Gln His Gly Val Ser Glu Glu Glu Ala Val Lys Glu Leu Leu Leu Glu 485 490 495 gtc gct aat tcc tgg aag gac atc aac gaa gaa ttg ttg aac cca acc 1536

Val Ala Asn Ser Trp Lys Asp Ile Asn Glu Glu Leu Leu Asn Pro Thr 500 505 510 act gtc cct ttg cct atg ttg caa aga ttg cta tac ttt gct aga tct 1584

Thr Val Pro Leu Pro Met Leu Gln Arg Leu Leu Tyr Phe Ala Arg Ser 515 520 525 ggc cac ttc atc tac gac gac ggt cat gat aga tac aca cac agt ttg 1632

Gly His Phe Ile Tyr Asp Asp Gly His Asp Arg Tyr Thr His Ser Leu 530 535 540 atg atg aaa aga caa gtt gct ttg tta tta acc gaa cca cta gct atc 1680

Met Met Lys Arg Gln Val Ala Leu Leu Leu Thr Glu Pro Leu Ala Ile 545 550 555 560 taa 1683 <210> 22 <211> 560 <212> PRT <213> Citrus junos <400> 22

Met Lys Asp Met Ser Ile Pro Leu Leu Ala Ala Val Ser Ser Ser Thr 1 5 10 15

Glu Glu Thr Val Arg Pro Ile Ala Asp Phe His Pro Thr Leu Trp Gly

Asn His Phe Leu Lys Ser Ala Ala Asp Val Glu Thr Ile Asp Ala Ala

Thr Gln Glu Gln His Ala Ala Leu Lys Gln Glu Val Arg Arg Met Ile 60

Thr Thr Thr Ala Asn Lys Leu Ala Gln Lys Leu His Met Ile Asp Ala

Pagina 56

SEQLTXT

65 70 75 80

Val Gln Arg Leu Gly Val Ala Tyr His Phe Glu Lys Glu Ile Glu Asp 85 90 95

Glu Leu Gly Lys Val Ser His Asp Leu Asp Ser Asp Asp Leu Tyr Val 100 105 110

Val Ser Leu Arg Phe Arg Leu Phe Arg Gln Gln Gly Val Lys Ile Ser 115 120 125

Cys Asp Val Phe Asp Lys Phe Lys Asp Asp Glu Gly Lys Phe Lys Glu 130 135 140

Ser Leu Ile Asn Asp Ile Arg Gly Met Leu Ser Leu Tyr Glu Ala Ala 145 150 155 160

Tyr Leu Ala Ile Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Val Phe 165 170 175

Thr Thr Thr His Leu Lys Ser Val Ile Ser Ile Ser Asp His Ser His 180 185 190

Ala Asn Ser Asn Leu Ala Glu Gln Ile Arg His Ser Leu Gln Ile Pro 195 200 205

Leu Arg Lys Ala Ala Ala Arg Leu Glu Ala Arg Tyr Phe Leu Asp Ile 210 215 220

Tyr Ser Arg Asp Asp Leu His Asp Glu Thr Leu Leu Lys Phe Ala Lys 225 230 235 240

Leu Asp Phe Asn Ile Leu Gln Ala Ala His Gln Lys Glu Ala Ser Ile 245 250 255

Met Thr Arg Trp Trp Asn Asp Leu Gly Phe Pro Lys Lys Val Pro Tyr 260 265 270

Ala Arg Asp Arg Ile Ile Glu Thr Tyr Ile Trp Met Leu Leu Gly Val

Pagina 57

SEQLTXT

275 280 285

Ser Tyr Glu Pro Asn Leu Ala Phe Gly Arg Ile Phe Ala Ser Lys Val 290 295 300

Val Cys Met Ile Thr Thr Ile Asp Asp Thr Phe Asp Ala Tyr Gly Thr 305 310 315 320

Phe Glu Glu Leu Thr Leu Phe Thr Glu Ala Val Thr Arg Trp Asp Ile 325 330 335

Gly Leu Ile Asp Thr Leu Pro Glu Tyr Met Lys Phe Ile Val Lys Ala 340 345 350

Leu Leu Asp Ile Tyr Arg Glu Ala Glu Glu Glu Leu Ala Lys Glu Gly 355 360 365

Arg Ser Tyr Gly Ile Pro Tyr Ala Lys Gln Met Met Gln Glu Leu Ile 370 375 380

Ile Leu Tyr Phe Thr Glu Ala Lys Trp Leu Tyr Lys Gly Tyr Val Pro 385 390 395 400

Thr Phe Asp Glu Tyr Lys Ser Val Ala Leu Arg Ser Ile Gly Leu Arg 405 410 415

Thr Leu Ala Val Ala Ser Phe Val Asp Leu Gly Asp Phe Ile Ala Thr 420 425 430

Lys Asp Asn Phe Glu Cys Ile Leu Lys Asn Ala Lys Ser Leu Lys Ala 435 440 445

Thr Glu Thr Ile Gly Arg Leu Met Asp Asp Ile Ala Gly Tyr Lys Phe 450 455 460

Glu Gln Lys Arg Gly His Asn Pro Ser Ala Val Glu Cys Tyr Lys Asn 465 470 475 480

Gln His Gly Val Ser Glu Glu Glu Ala Val Lys Glu Leu Leu Leu Glu

Pagina 58

SEQLTXT

485 490 495

Val Ala Asn Ser Trp Lys Asp Ile Asn Glu Glu Leu Leu Asn Pro Thr 500 505 510

Thr Val Pro Leu Pro Met Leu Gln Arg Leu Leu Tyr Phe Ala Arg Ser 515 520 525

Gly His Phe Ile Tyr Asp Asp Gly His Asp Arg Tyr Thr His Ser Leu 530 535 540

Met Met Lys Arg Gln Val Ala Leu Leu Leu Thr Glu Pro Leu Ala Ile 545 550 555 560 <21e> 23 <211> 1683 <212> DNA <213> Citrus junos <220> <221> CDS <222> (1)..(1683) <400> 23 atg aaa gat atg tca ata cct ttg ctt gca gct gtc tct tect tca acc 48

Met Lys Asp Met Ser Ile Pro Leu Leu Ala Ala Val Ser Ser Ser Thr 1 5 10 15 gaa gaa aca gtt aga cca ata gct gac ttc cat cct act ttg tgg gga 96

Glu Glu Thr Val Arg Pro Ile Ala Asp Phe His Pro Thr Leu Trp Gly aat cat ttt ctt aag agc gct gcc gat gtc gag aca ata gac gct gct 144

Asn His Phe Leu Lys Ser Ala Ala Asp Val Glu Thr Ile Asp Ala Ala aca Caa gaa caa cac gca gca ctt aaa cag gaa gtt aga cgt atg atc 192

Thr Gln Glu Gln His Ala Ala Leu Lys Gln Glu Val Arg Arg Met Ile 60 act aca act gcg aat aaa ttg gcc caa aaa tta cat atg atc gac gcc 240

Thr Thr Thr Ala Asn Lys Leu Ala Gln Lys Leu His Met Ile Asp Ala 65 70 75 80 gtt cag aga cta ggg gta gct tac cac ttc gag aaa gag ata gaa gat 288

Val Gln Arg Leu Gly Val Ala Tyr His Phe Glu Lys Glu Ile Glu Asp

Pagina 59

SEQLTXT

85 90 95 gag ttg ggc aaa gta tca cac gat ctt gac agc gat gat ctg tac gtg 336

Glu Leu Gly Lys Val Ser His Asp Leu Asp Ser Asp Asp Leu Tyr Val 100 105 110 gtt tcc ctt agg ttc aga ctg ttt aga cag cag gga gtt aaa atc agc 384

Val Ser Leu Arg Phe Arg Leu Phe Arg Gln Gln Gly Val Lys Ile Ser 115 120 125 tgt gat gtt ttt gat aaa ttt aaa gat gat gaa ggc aaa ttt aaa gaa 432

Cys Asp Val Phe Asp Lys Phe Lys Asp Asp Glu Gly Lys Phe Lys Glu 130 135 140 tct ctt atc aat gac atc aga ggc atg cta agt ttg tat gag gcg gcc 480

Ser Leu Ile Asn Asp Ile Arg Gly Met Leu Ser Leu Tyr Glu Ala Ala 145 150 155 160 tat ctg gca att aga ggc gag gat att ttg gat gaa gca ata gtt ttc 528

Tyr Leu Ala Ile Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Val Phe 165 170 175 aca acc aca cat tta aag tcc gtt att agc att tct gat cat agc cat 576

Thr Thr Thr His Leu Lys Ser Val Ile Ser Ile Ser Asp His Ser His 180 185 190 gca aat tcg aac tta gct gaa cag atc agg cat agc cta caa att cca 624

Ala Asn Ser Asn Leu Ala Glu Gln Ile Arg His Ser Leu Gln Ile Pro 195 200 205 ttg agg aag gct gcc gcc aga tta gag gct agg tat ttt tta gat att 672

Leu Arg Lys Ala Ala Ala Arg Leu Glu Ala Arg Tyr Phe Leu Asp Ile 210 215 220 tac tca aga gat gat ctg cat gat gaa act tta ctg aaa ttc gca aag 720

Tyr Ser Arg Asp Asp Leu His Asp Glu Thr Leu Leu Lys Phe Ala Lys 225 230 235 240 tta gac ttt aat att ttg caa gcc gcg cat caa aaa gaa gct tcg att 768

Leu Asp Phe Asn Ile Leu Gln Ala Ala His Gln Lys Glu Ala Ser Ile 245 250 255 atg aca aga tgg tgg aat gat tta ggt ttt ccg aaa aaa gtc cca tac 816

Met Thr Arg Trp Trp Asn Asp Leu Gly Phe Pro Lys Lys Val Pro Tyr 260 265 270 gcg aga gac agg atc atc gag act tat att tgg atg tta tta ggg gtt 864

Ala Arg Asp Arg Ile Ile Glu Thr Tyr Ile Trp Met Leu Leu Gly Val 275 280 285 tca tac gag cct aac ctt gct ttt ggt aga atc ttc gct tca aaa gtt 912

Ser Tyr Glu Pro Asn Leu Ala Phe Gly Arg Ile Phe Ala Ser Lys Val

Pagina 60

SEQLTXT

290 295 300 gtt tgt atg ata acc act ata gac gat aca ttc gat gca tac ggt act 960

Val Cys Met Ile Thr Thr Ile Asp Asp Thr Phe Asp Ala Tyr Gly Thr 305 310 315 320 ttc gaa gaa cta aca cta ttc acg gaa gcc gtg aca agg tgg gat ata 1008

Phe Glu Glu Leu Thr Leu Phe Thr Glu Ala Val Thr Arg Trp Asp Ile 325 330 335 gga ttg ata gat acc cta cct gaa tat atg aag ttt att gtg aaa gcc 1056

Gly Leu Ile Asp Thr Leu Pro Glu Tyr Met Lys Phe Ile Val Lys Ala 340 345 350 tta ttg gac att tat aga gaa gca gaa gaa gaa ttg gcc aaa gag ggc 1104

Leu Leu Asp Ile Tyr Arg Glu Ala Glu Glu Glu Leu Ala Lys Glu Gly 355 360 365 aga tct tac ggg att ccc tac gct aaa caa atg atg caa gag ctt ata 1152

Arg Ser Tyr Gly Ile Pro Tyr Ala Lys Gln Met Met Gln Glu Leu Ile 370 375 380 att ttg tac ttc aca gag gct aaa tgg cta tat aag gga tac gtc ccg 1200

Ile Leu Tyr Phe Thr Glu Ala Lys Trp Leu Tyr Lys Gly Tyr Val Pro 385 390 395 400 acc ttt gac gaa tac aaa tcg gtc gca ctt cgt agc att ggg ttg aga 1248

Thr Phe Asp Glu Tyr Lys Ser Val Ala Leu Arg Ser Ile Gly Leu Arg 405 410 415 aca tta gcg gtg gca tct ttt gtt gac tta ggg gat ttt att gct aca 1296

Thr Leu Ala Val Ala Ser Phe Val Asp Leu Gly Asp Phe Ile Ala Thr 420 425 430 aaa gac aac ttc gaa tgt att tta aaa aat gcc aaa agc ttg aaa gcg 1344

Lys Asp Asn Phe Glu Cys Ile Leu Lys Asn Ala Lys Ser Leu Lys Ala 435 440 445 act gag acc att ggt agg cta atg gat gat att gca ggt tat aaa ttt 1392

Thr Glu Thr Ile Gly Arg Leu Met Asp Asp Ile Ala Gly Tyr Lys Phe 450 455 460 gaa caa aaa aga ggt cat aac cct tct gct gtt gaa tgt tac aag aat 14409

Glu Gln Lys Arg Gly His Asn Pro Ser Ala Val Glu Cys Tyr Lys Asn 465 470 475 480 cag cat ggt gtt agt gaa gag gag gca gtg aaa gaa ttg ctt ttg gaa 1488

Gln His Gly Val Ser Glu Glu Glu Ala Val Lys Glu Leu Leu Leu Glu 485 490 495 gtc gcg aat tct tgg aag gac att aat gag gag cta ctt aac cca acc 1536

Val Ala Asn Ser Trp Lys Asp Ile Asn Glu Glu Leu Leu Asn Pro Thr

Pagina 61

SEQLTXT

500 505 510 acg gtt cct ttg ccg atg ttg caa aga tta ttg tat ttc gct aga agc 1584

Thr Val Pro Leu Pro Met Leu Gln Arg Leu Leu Tyr Phe Ala Arg Ser 515 520 525 ggt cat ttc ata tat gat gat gga cat gat aga tat act cat agt tta 1632

Gly His Phe Ile Tyr Asp Asp Gly His Asp Arg Tyr Thr His Ser Leu 530 535 540 atg atg aag agg cag gta gcc cta ctt ttg acg gaa cct cta gca att 1680

Met Met Lys Arg Gln Val Ala Leu Leu Leu Thr Glu Pro Leu Ala Ile 545 550 555 560 tga 1683 <210> 24 <211> 560 <212> PRT <213> Citrus junos <400> 24

Met Lys Asp Met Ser Ile Pro Leu Leu Ala Ala Val Ser Ser Ser Thr 1 5 10 15

Glu Glu Thr Val Arg Pro Ile Ala Asp Phe His Pro Thr Leu Trp Gly

Asn His Phe Leu Lys Ser Ala Ala Asp Val Glu Thr Ile Asp Ala Ala

Thr Gln Glu Gln His Ala Ala Leu Lys Gln Glu Val Arg Arg Met Ile 60

Thr Thr Thr Ala Asn Lys Leu Ala Gln Lys Leu His Met Ile Asp Ala 65 70 75 80

Val Gln Arg Leu Gly Val Ala Tyr His Phe Glu Lys Glu Ile Glu Asp 85 90 95

Glu Leu Gly Lys Val Ser His Asp Leu Asp Ser Asp Asp Leu Tyr Val 100 105 119

Pagina 62

SEQLTXT

Val Ser Leu Arg Phe Arg Leu Phe Arg Gln Gln Gly Val Lys Ile Ser 115 120 125

Cys Asp Val Phe Asp Lys Phe Lys Asp Asp Glu Gly Lys Phe Lys Glu 130 135 140

Ser Leu Ile Asn Asp Ile Arg Gly Met Leu Ser Leu Tyr Glu Ala Ala 145 150 155 160

Tyr Leu Ala Ile Arg Gly Glu Asp Ile Leu Asp Glu Ala Ile Val Phe 165 170 175

Thr Thr Thr His Leu Lys Ser Val Ile Ser Ile Ser Asp His Ser His 180 185 190

Ala Asn Ser Asn Leu Ala Glu Gln Ile Arg His Ser Leu Gln Ile Pro 195 200 205

Leu Arg Lys Ala Ala Ala Arg Leu Glu Ala Arg Tyr Phe Leu Asp Ile 210 215 220

Tyr Ser Arg Asp Asp Leu His Asp Glu Thr Leu Leu Lys Phe Ala Lys 225 230 235 240

Leu Asp Phe Asn Ile Leu Gln Ala Ala His Gln Lys Glu Ala Ser Ile 245 250 255

Met Thr Arg Trp Trp Asn Asp Leu Gly Phe Pro Lys Lys Val Pro Tyr 260 265 270

Ala Arg Asp Arg Ile Ile Glu Thr Tyr Ile Trp Met Leu Leu Gly Val 275 280 285

Ser Tyr Glu Pro Asn Leu Ala Phe Gly Arg Ile Phe Ala Ser Lys Val 290 295 300

Val Cys Met Ile Thr Thr Ile Asp Asp Thr Phe Asp Ala Tyr Gly Thr 305 310 315 320

Pagina 63

SEQLTXT

Phe Glu Glu Leu Thr Leu Phe Thr Glu Ala Val Thr Arg Trp Asp Ile 325 330 335

Gly Leu Ile Asp Thr Leu Pro Glu Tyr Met Lys Phe Ile Val Lys Ala 340 345 350

Leu Leu Asp Ile Tyr Arg Glu Ala Glu Glu Glu Leu Ala Lys Glu Gly 355 360 365

Arg Ser Tyr Gly Ile Pro Tyr Ala Lys Gln Met Met Gln Glu Leu Ile 370 375 380

Ile Leu Tyr Phe Thr Glu Ala Lys Trp Leu Tyr Lys Gly Tyr Val Pro 385 390 395 400

Thr Phe Asp Glu Tyr Lys Ser Val Ala Leu Arg Ser Ile Gly Leu Arg 405 410 415

Thr Leu Ala Val Ala Ser Phe Val Asp Leu Gly Asp Phe Ile Ala Thr 420 425 430

Lys Asp Asn Phe Glu Cys Ile Leu Lys Asn Ala Lys Ser Leu Lys Ala 435 440 445

Thr Glu Thr Ile Gly Arg Leu Met Asp Asp Ile Ala Gly Tyr Lys Phe 450 455 460

Glu Gln Lys Arg Gly His Asn Pro Ser Ala Val Glu Cys Tyr Lys Asn 465 470 475 480

Gln His Gly Val Ser Glu Glu Glu Ala Val Lys Glu Leu Leu Leu Glu 485 490 495

Val Ala Asn Ser Trp Lys Asp Ile Asn Glu Glu Leu Leu Asn Pro Thr 500 505 510

Thr Val Pro Leu Pro Met Leu Gln Arg Leu Leu Tyr Phe Ala Arg Ser 515 520 525

Pagina 64

SEQLTXT

Gly His Phe Ile Tyr Asp Asp Gly His Asp Arg Tyr Thr His Ser Leu 530 535 540

Met Met Lys Arg Gln Val Ala Leu Leu Leu Thr Glu Pro Leu Ala Ile 545 550 555 560 <210> 25 <211> 1725 <212> DNA <213> Matricaria chamomilla var. recutita <220> <221> CDS <222> (1)..(1725) <400> 25 atg agc acg att cca gtg tca tca gta tcg ttt tca tca tcc gcc agt 48

Met Ser Thr Ile Pro Val Ser Ser Val Ser Phe Ser Ser Ser Ala Ser 1 5 10 15 ccg ctt gtt ctt gat gac aaa ctt tcg acg aag cag gac gtc gtc aga 96

Pro Leu Val Leu Asp Asp Lys Leu Ser Thr Lys Gln Asp Val Val Arg cat acg atg aac ttc tct gct agt att tgg ggt gac caa ttt ttg act 144

His Thr Met Asn Phe Ser Ala Ser Ile Trp Gly Asp Gln Phe Leu Thr tac cat gag cca gaa gac tta gta att aaa aaa caa cag gtt gag cag 192

Tyr His Glu Pro Glu Asp Leu Val Ile Lys Lys Gln Gln Val Glu Gln 60 ctg aaa gag gaa gtt aaa aag gaa ttg atg gcc ata aaa ggc tcc aac 240

Leu Lys Glu Glu Val Lys Lys Glu Leu Met Ala Ile Lys Gly Ser Asn 65 70 75 80 gac cct caa cag cac att aaa ctt atg gaa ttg att gac tca gtt caa 288

Asp Pro Gln Gln His Ile Lys Leu Met Glu Leu Ile Asp Ser Val Gln 85 90 95 agg tta ggt att gca tat cat ttt gag gaa gaa att gaa gaa gcc ctt 336

Arg Leu Gly Ile Ala Tyr His Phe Glu Glu Glu Ile Glu Glu Ala Leu 100 105 110 caa cac att cat gtg act tat ggg gaa cat tgg gtt gac aaa gaa aac 384

Gln His Ile His Val Thr Tyr Gly Glu His Trp Val Asp Lys Glu Asn 115 120 125 tta cag agc gtg tcg tta tgg ttt aga ttg ctg aga cag caa ggc ttt 432

Pagina 65

SEQLTXT

Leu Gln Ser Val Ser Leu Trp Phe Arg Leu Leu Arg Gln Gln Gly Phe 130 135 140 aat gtc tct tcc ggt gta ttt aag gat tat atg gat gaa aag ggt aac 480

Asn Val Ser Ser Gly Val Phe Lys Asp Tyr Met Asp Glu Lys Gly Asn 145 150 155 160 ttc aaa gag tcc ctt tgc aac gac gcc cag ggt att ctg get ttg tac 528

Phe Lys Glu Ser Leu Cys Asn Asp Ala Gln Gly Ile Leu Ala Leu Tyr 165 170 175 gaa gct gca tat atg agg gtc gaa gga gaa act agg ttg gat aag gca 576

Glu Ala Ala Tyr Met Arg Val Glu Gly Glu Thr Arg Leu Asp Lys Ala 180 185 190 ttg gaa ttt aca aaa gtc cac tta gac att att tca aag gat ccg tcc 624

Leu Glu Phe Thr Lys Val His Leu Asp Ile Ile Ser Lys Asp Pro Ser 195 200 205 tgt gac tct tac tta cgt aca agg gtc cat caa gca ttg aag caa cca 672

Cys Asp Ser Tyr Leu Arg Thr Arg Val His Gln Ala Leu Lys Gln Pro 210 215 220 ttg agg aga agg cta gct aga att gag gct cta cat tat atg cca gta 720

Leu Arg Arg Arg Leu Ala Arg Ile Glu Ala Leu His Tyr Met Pro Val 225 230 235 240 tac caa caa gat tct agt cat aat gaa gtt tta tta aaa ttg gcg aag 768

Tyr Gln Gln Asp Ser Ser His Asn Glu Val Leu Leu Lys Leu Ala Lys 245 250 255 gta gac ttc agc gtc ctg cag tct atg cat aaa aag gaa ctt agc cac 816

Val Asp Phe Ser Val Leu Gln Ser Met His Lys Lys Glu Leu Ser His 260 265 270 att tgt aag tgg tgg aaa gat ctt gac tta caa aac aaa tta cct tat 864

Ile Cys Lys Trp Trp Lys Asp Leu Asp Leu Gln Asn Lys Leu Pro Tyr 275 280 285 gta agg gat aga gtc gtt gaa ggc tat ttt tgg atc tta tca att tat 912

Val Arg Asp Arg Val Val Glu Gly Tyr Phe Trp Ile Leu Ser Ile Tyr 290 295 300 tat gaa ccg cag cat gct agg aca aga atg ttt ttg atg aag act tgt 960

Tyr Glu Pro Gln His Ala Arg Thr Arg Met Phe Leu Met Lys Thr Cys 305 310 315 320 atg tgg ttg gta gtt ttg gat gat acg ttc gat aat tat gga aca tat 1008

Met Trp Leu Val Val Leu Asp Asp Thr Phe Asp Asn Tyr Gly Thr Tyr 325 330 335 gaa gaa tta gaa atc ttc gcg caa gcc gtt gaa cgt tgg agt att tcc 1056

Pagina 66

SEQLTXT

Glu Glu Leu Glu Ile Phe Ala Gln Ala Val Glu Arg Trp Ser Ile Ser 340 345 350 tgc atg gat atg ctg cct gag tat atg aaa tta att tat cag gaa ttg 1104

Cys Met Asp Met Leu Pro Glu Tyr Met Lys Leu Ile Tyr Gln Glu Leu 355 360 365 gta aat ctt cat gta gac atg gaa gaa tca tta gag aaa gaa gga aaa 1152

Val Asn Leu His Val Asp Met Glu Glu Ser Leu Glu Lys Glu Gly Lys 370 375 380 acc tat caa att cac tat gtc aaa gaa atg gcc aaa gag tta gtc agg 1200

Thr Tyr Gln Ile His Tyr Val Lys Glu Met Ala Lys Glu Leu Val Arg 385 390 395 400 aac tat tta gta gaa gcg agg tgg cta aag gaa ggg tac atg ccg aca 1248

Asn Tyr Leu Val Glu Ala Arg Trp Leu Lys Glu Gly Tyr Met Pro Thr 405 410 415 ttg gaa gag tac atg tcc gtt tct atg gtt aca ggg acc tat ggc ctt 1296

Leu Glu Glu Tyr Met Ser Val Ser Met Val Thr Gly Thr Tyr Gly Leu 420 425 430 atg att gcc agg tca tat gta gga aga gat gat att gtg aca gag gat 1344

Met Ile Ala Arg Ser Tyr Val Gly Arg Asp Asp Ile Val Thr Glu Asp 435 440 445 aca ttt aag tgg gtt tca tcc tat cct ccc att att aaa gcg tcc tgt 1392

Thr Phe Lys Trp Val Ser Ser Tyr Pro Pro Ile Ile Lys Ala Ser Cys 450 455 460 gtt atc gtt cgt cta atg gat gac att gtt tct cac aaa gaa gaa cag 14409

Val Ile Val Arg Leu Met Asp Asp Ile Val Ser His Lys Glu Glu Gln 465 470 475 480 gaa aga gga cat gtt gca agt tca att gaa tgc tat tcg aaa gag tcc 1488

Glu Arg Gly His Val Ala Ser Ser Ile Glu Cys Tyr Ser Lys Glu Ser 485 490 495 ggt gca aca gag gaa gaa gct tgt gaa tac att tct tca aag gtt gaa 1536

Gly Ala Thr Glu Glu Glu Ala Cys Glu Tyr Ile Ser Ser Lys Val Glu 500 505 510 gat gcc tgg aag gtc ata aat aga gaa tct ctg agg ccg acc gct gtc 1584

Asp Ala Trp Lys Val Ile Asn Arg Glu Ser Leu Arg Pro Thr Ala Val 515 520 525 cca ttt ccc ttg ctt atg cct gcg ata aat cta gca aga atg tgt gag 1632

Pro Phe Pro Leu Leu Met Pro Ala Ile Asn Leu Ala Arg Met Cys Glu 530 535 540 gtc cta tat agt gtg aac gac ggt ttt aca cat gca gaa ggt gat atg 1680

Pagina 67

SEQLTXT

Val Leu Tyr Ser Val Asn Asp Gly Phe Thr His Ala Glu Gly Asp Met 545 550 555 560 aaa tcg tac atg aaa agt tat ttc gtc cac cct atg gtg att taa 1725

Lys Ser Tyr Met Lys Ser Tyr Phe Val His Pro Met Val Ile 565 570 <210> 26 <211> 574 <212> PRT <213> Matricaria chamomilla var. recutita <400> 26

Met Ser Thr Ile Pro Val Ser Ser Val Ser Phe Ser Ser Ser Ala Ser 1 5 10 15

Pro Leu Val Leu Asp Asp Lys Leu Ser Thr Lys Gln Asp Val Val Arg

His Thr Met Asn Phe Ser Ala Ser Ile Trp Gly Asp Gln Phe Leu Thr

Tyr His Glu Pro Glu Asp Leu Val Ile Lys Lys Gln Gln Val Glu Gln 60

Leu Lys Glu Glu Val Lys Lys Glu Leu Met Ala Ile Lys Gly Ser Asn 65 70 75 80

Asp Pro Gln Gln His Ile Lys Leu Met Glu Leu Ile Asp Ser Val Gln 85 90 95

Arg Leu Gly Ile Ala Tyr His Phe Glu Glu Glu Ile Glu Glu Ala Leu 100 105 119

Gln His Ile His Val Thr Tyr Gly Glu His Trp Val Asp Lys Glu Asn 115 120 125

Leu Gln Ser Val Ser Leu Trp Phe Arg Leu Leu Arg Gln Gln Gly Phe 130 135 140

Asn Val Ser Ser Gly Val Phe Lys Asp Tyr Met Asp Glu Lys Gly Asn

Pagina 68

SEQLTXT

145 150 155 160

Phe Lys Glu Ser Leu Cys Asn Asp Ala Gln Gly Ile Leu Ala Leu Tyr 165 170 175

Glu Ala Ala Tyr Met Arg Val Glu Gly Glu Thr Arg Leu Asp Lys Ala 180 185 190

Leu Glu Phe Thr Lys Val His Leu Asp Ile Ile Ser Lys Asp Pro Ser 195 200 205

Cys Asp Ser Tyr Leu Arg Thr Arg Val His Gln Ala Leu Lys Gln Pro 210 215 220

Leu Arg Arg Arg Leu Ala Arg Ile Glu Ala Leu His Tyr Met Pro Val 225 230 235 240

Tyr Gln Gln Asp Ser Ser His Asn Glu Val Leu Leu Lys Leu Ala Lys 245 250 255

Val Asp Phe Ser Val Leu Gln Ser Met His Lys Lys Glu Leu Ser His 260 265 270

Ile Cys Lys Trp Trp Lys Asp Leu Asp Leu Gln Asn Lys Leu Pro Tyr 275 280 285

Val Arg Asp Arg Val Val Glu Gly Tyr Phe Trp Ile Leu Ser Ile Tyr 290 295 300

Tyr Glu Pro Gln His Ala Arg Thr Arg Met Phe Leu Met Lys Thr Cys 305 310 315 320

Met Trp Leu Val Val Leu Asp Asp Thr Phe Asp Asn Tyr Gly Thr Tyr 325 330 335

Glu Glu Leu Glu Ile Phe Ala Gln Ala Val Glu Arg Trp Ser Ile Ser 340 345 350

Cys Met Asp Met Leu Pro Glu Tyr Met Lys Leu Ile Tyr Gln Glu Leu

Pagina 69

SEQLTXT

355 360 365

Val Asn Leu His Val Asp Met Glu Glu Ser Leu Glu Lys Glu Gly Lys 370 375 380

Thr Tyr Gln Ile His Tyr Val Lys Glu Met Ala Lys Glu Leu Val Arg 385 390 395 400

Asn Tyr Leu Val Glu Ala Arg Trp Leu Lys Glu Gly Tyr Met Pro Thr 405 410 415

Leu Glu Glu Tyr Met Ser Val Ser Met Val Thr Gly Thr Tyr Gly Leu 420 425 430

Met Ile Ala Arg Ser Tyr Val Gly Arg Asp Asp Ile Val Thr Glu Asp 435 440 445

Thr Phe Lys Trp Val Ser Ser Tyr Pro Pro Ile Ile Lys Ala Ser Cys 450 455 460

Val Ile Val Arg Leu Met Asp Asp Ile Val Ser His Lys Glu Glu Gln 465 470 475 480

Glu Arg Gly His Val Ala Ser Ser Ile Glu Cys Tyr Ser Lys Glu Ser 485 490 495

Gly Ala Thr Glu Glu Glu Ala Cys Glu Tyr Ile Ser Ser Lys Val Glu 500 505 510

Asp Ala Trp Lys Val Ile Asn Arg Glu Ser Leu Arg Pro Thr Ala Val 515 520 525

Pro Phe Pro Leu Leu Met Pro Ala Ile Asn Leu Ala Arg Met Cys Glu 530 535 540

Val Leu Tyr Ser Val Asn Asp Gly Phe Thr His Ala Glu Gly Asp Met 545 550 555 560

Lys Ser Tyr Met Lys Ser Tyr Phe Val His Pro Met Val Ile

Pagina 70

SEQLTXT

565 570 <210> 27 <211> 1653 <212> DNA <213> Mentha piperita <220> <221> CDS <222> (1)..(1653) <400> 27 atg gca act aat ggc gtc gtg atc tca tgt cta agg gag gtc aga cca 48

Met Ala Thr Asn Gly Val Val Ile Ser Cys Leu Arg Glu Val Arg Pro 1 5 10 15 cct atg aca aaa cat gcc ccc agc atg tgg aca gat acc ttt tcg aac 96

Pro Met Thr Lys His Ala Pro Ser Met Trp Thr Asp Thr Phe Ser Asn ttc tcg tta gat gat aag gag caa caa aaa tgc tct gaa acc att gag 144

Phe Ser Leu Asp Asp Lys Glu Gln Gln Lys Cys Ser Glu Thr Ile Glu gct tta aaa caa gaa gct aga gga atg tta atg gct gca aca aca ccc 192

Ala Leu Lys Gln Glu Ala Arg Gly Met Leu Met Ala Ala Thr Thr Pro 60 ctt caa caa atg acg tta ata gat aca ctg gaa aga ctg ggg tta tct 240

Leu Gln Gln Met Thr Leu Ile Asp Thr Leu Glu Arg Leu Gly Leu Ser 65 70 75 80 ttt cat ttt gaa acc gaa att gag tat aag att gag ctg att aac gca 288

Phe His Phe Glu Thr Glu Ile Glu Tyr Lys Ile Glu Leu Ile Asn Ala 85 90 95 gct gaa gac gat ggc ttc gat ttg ttt gcg aca gct ttg cgt ttc aga 336

Ala Glu Asp Asp Gly Phe Asp Leu Phe Ala Thr Ala Leu Arg Phe Arg 100 105 110 ttg ctt agg caa cac cag aga cat gta tcc tgt gac gtt ttc gat aag 384

Leu Leu Arg Gln His Gln Arg His Val Ser Cys Asp Val Phe Asp Lys 115 120 125 ttt att gac aaa gat gga aaa ttt gaa gag agc ctt tct aat aat gta 432

Phe Ile Asp Lys Asp Gly Lys Phe Glu Glu Ser Leu Ser Asn Asn Val 130 135 140 gaa ggt ttg cta agc tta tat gaa gct gca cac gtc gga ttt aga gag 480

Glu Gly Leu Leu Ser Leu Tyr Glu Ala Ala His Val Gly Phe Arg Glu

Pagina 71

SEQLTXT

145 150 155 160 gag aga att ttg caa gaa gct gta aat ttt aca cgt cac cac ttg gag 528

Glu Arg Ile Leu Gln Glu Ala Val Asn Phe Thr Arg His His Leu Glu 165 170 175 ggt gca gag tta gac cag tcg cca ttg ctg ata aga gag aaa gtg aag 576

Gly Ala Glu Leu Asp Gln Ser Pro Leu Leu Ile Arg Glu Lys Val Lys 180 185 190 aga gcc tta gaa cat cca ctg cat agg gat ttc cca att gtt tac gct 624

Arg Ala Leu Glu His Pro Leu His Arg Asp Phe Pro Ile Val Tyr Ala 195 200 205 aga ttg ttt att agt att tac gaa aaa gac gat tct cgt gat gaa ctt 672

Arg Leu Phe Ile Ser Ile Tyr Glu Lys Asp Asp Ser Arg Asp Glu Leu 210 215 220 ctt tta aag ttg agc aaa gtc aat ttt aaa ttt atg caa aat ctt tac 720

Leu Leu Lys Leu Ser Lys Val Asn Phe Lys Phe Met Gln Asn Leu Tyr 225 230 235 240 aaa gaa gag ctg tct caa ttg tct agg tgg tgg aac act tgg aat tta 768

Lys Glu Glu Leu Ser Gln Leu Ser Arg Trp Trp Asn Thr Trp Asn Leu 245 250 255 aag agc aag tta ccc tat gct cgt gat agg gta gtc gaa gct tat gtt 816

Lys Ser Lys Leu Pro Tyr Ala Arg Asp Arg Val Val Glu Ala Tyr Val 260 265 270 tgg ggt gtt ggc tat cac tac gag cct cag tac tca tac gtt aga atg 864

Trp Gly Val Gly Tyr His Tyr Glu Pro Gln Tyr Ser Tyr Val Arg Met 275 280 285 ggt cta gct aag ggt gta tta att tgt ggg ata atg gat gat acc tat 912

Gly Leu Ala Lys Gly Val Leu Ile Cys Gly Ile Met Asp Asp Thr Tyr 290 295 300 gac aac tat gcc aca tta aac gag gct caa ttg ttc aca caa gtc tta 960

Asp Asn Tyr Ala Thr Leu Asn Glu Ala Gln Leu Phe Thr Gln Val Leu 305 310 315 320 gat aag tgg gat aga gat gag gca gaa aga ttg ccg gaa tac atg aaa 1008

Asp Lys Trp Asp Arg Asp Glu Ala Glu Arg Leu Pro Glu Tyr Met Lys 325 330 335 atc gta tat cgt ttt att tta tct att tat gag aac tac gaa cgt gac 1056

Ile Val Tyr Arg Phe Ile Leu Ser Ile Tyr Glu Asn Tyr Glu Arg Asp 340 345 350 gcc gct aaa tta ggc aag tca ttt gcg gcc cct tac ttt aaa gaa act 1104

Ala Ala Lys Leu Gly Lys Ser Phe Ala Ala Pro Tyr Phe Lys Glu Thr

Pagina 72

SEQLTXT

355 360 365 gtc aaa caa cta gct aga gca ttt aac gag gag caa aag tgg gtt atg 1152

Val Lys Gln Leu Ala Arg Ala Phe Asn Glu Glu Gln Lys Trp Val Met 370 375 380 gag aga cag tta cca tcg ttt caa gat tat gtc aag aac tcc gag aag 1200

Glu Arg Gln Leu Pro Ser Phe Gln Asp Tyr Val Lys Asn Ser Glu Lys 385 390 395 400 act tcc tgc atc tac aca atg ttt gct tct ata att cct ggc ttg aaa 1248

Thr Ser Cys Ile Tyr Thr Met Phe Ala Ser Ile Ile Pro Gly Leu Lys 405 410 415 agc gtg act cag gag aca att gat tgg ata aag tcc gaa cca act ttg 1296

Ser Val Thr Gln Glu Thr Ile Asp Trp Ile Lys Ser Glu Pro Thr Leu 420 425 430 gcg acc tca acg gct atg atc ggc aga tac tgg aat gat act agt tcg 1344

Ala Thr Ser Thr Ala Met Ile Gly Arg Tyr Trp Asn Asp Thr Ser Ser 435 440 445 caa ttg aga gag agt aag gga gga gaa atg ttg act gca ctg gac ttt 1392

Gln Leu Arg Glu Ser Lys Gly Gly Glu Met Leu Thr Ala Leu Asp Phe 450 455 460 cat atg aag gaa tac ggg tta aca aaa gag gag gca gca tca aaa ttt 14409

His Met Lys Glu Tyr Gly Leu Thr Lys Glu Glu Ala Ala Ser Lys Phe 465 470 475 480 gaa ggt ctt gtc gaa gaa act tgg aaa gat atc aac aaa gag ttc ata 1488

Glu Gly Leu Val Glu Glu Thr Trp Lys Asp Ile Asn Lys Glu Phe Ile 485 490 495 gca acg acg aat tat aac gta gga aga gag att gca att act ttt ctt 1536

Ala Thr Thr Asn Tyr Asn Val Gly Arg Glu Ile Ala Ile Thr Phe Leu 500 505 510 aat tat gca agg atc tgc gaa gcc agt tat agt aaa act gac ggg gac 1584

Asn Tyr Ala Arg Ile Cys Glu Ala Ser Tyr Ser Lys Thr Asp Gly Asp 515 520 525 gcc tac tcg gat ccg aat gta gca aaa gct aat gtt gtg gct ttg ttt 1632

Ala Tyr Ser Asp Pro Asn Val Ala Lys Ala Asn Val Val Ala Leu Phe 530 535 540 gtt gac gca atc gta ttt tag 1653

Val Asp Ala Ile Val Phe 545 550 <210> 28

Pagina 73

SEQLTXT

<211> 559 <212> PRT <213> Mentha piperita <400> 28

Met Ala Thr Asn Gly Val Val Ile Ser Cys Leu Arg Glu Val Arg Pro 1 5 10 15

Pro Met Thr Lys His Ala Pro Ser Met Trp Thr Asp Thr Phe Ser Asn

Phe Ser Leu Asp Asp Lys Glu Gln Gln Lys Cys Ser Glu Thr Ile Glu

Ala Leu Lys Gln Glu Ala Arg Gly Met Leu Met Ala Ala Thr Thr Pro 60

Leu Gln Gln Met Thr Leu Ile Asp Thr Leu Glu Arg Leu Gly Leu Ser 65 70 75 80

Phe His Phe Glu Thr Glu Ile Glu Tyr Lys Ile Glu Leu Ile Asn Ala 85 90 95

Ala Glu Asp Asp Gly Phe Asp Leu Phe Ala Thr Ala Leu Arg Phe Arg 100 105 110

Leu Leu Arg Gln His Gln Arg His Val Ser Cys Asp Val Phe Asp Lys 115 120 125

Phe Ile Asp Lys Asp Gly Lys Phe Glu Glu Ser Leu Ser Asn Asn Val 130 135 140

Glu Gly Leu Leu Ser Leu Tyr Glu Ala Ala His Val Gly Phe Arg Glu 145 150 155 160

Glu Arg Ile Leu Gln Glu Ala Val Asn Phe Thr Arg His His Leu Glu 165 170 175

Gly Ala Glu Leu Asp Gln Ser Pro Leu Leu Ile Arg Glu Lys Val Lys 180 185 190

Pagina 74

SEQLTXT

Arg Ala Leu Glu His Pro Leu His Arg Asp Phe Pro Ile Val Tyr Ala 195 200 205

Arg Leu Phe Ile Ser Ile Tyr Glu Lys Asp Asp Ser Arg Asp Glu Leu 210 215 220

Leu Leu Lys Leu Ser Lys Val Asn Phe Lys Phe Met Gln Asn Leu Tyr 225 230 235 240

Lys Glu Glu Leu Ser Gln Leu Ser Arg Trp Trp Asn Thr Trp Asn Leu 245 250 255

Lys Ser Lys Leu Pro Tyr Ala Arg Asp Arg Val Val Glu Ala Tyr Val 260 265 270

Trp Gly Val Gly Tyr His Tyr Glu Pro Gln Tyr Ser Tyr Val Arg Met 275 280 285

Gly Leu Ala Lys Gly Val Leu Ile Cys Gly Ile Met Asp Asp Thr Tyr 290 295 300

Asp Asn Tyr Ala Thr Leu Asn Glu Ala Gln Leu Phe Thr Gln Val Leu 305 310 315 320

Asp Lys Trp Asp Arg Asp Glu Ala Glu Arg Leu Pro Glu Tyr Met Lys 325 330 335

Ile Val Tyr Arg Phe Ile Leu Ser Ile Tyr Glu Asn Tyr Glu Arg Asp 340 345 350

Ala Ala Lys Leu Gly Lys Ser Phe Ala Ala Pro Tyr Phe Lys Glu Thr 355 360 365

Val Lys Gln Leu Ala Arg Ala Phe Asn Glu Glu Gln Lys Trp Val Met 370 375 380

Glu Arg Gln Leu Pro Ser Phe Gln Asp Tyr Val Lys Asn Ser Glu Lys 385 390 395 400

Pagina 75

SEQLTXT

Thr Ser Cys Ile Tyr Thr Met Phe Ala Ser Ile Ile Pro Gly Leu Lys 405 410 415

Ser Val Thr Gln Glu Thr Ile Asp Trp Ile Lys Ser Glu Pro Thr Leu 420 425 430

Ala Thr Ser Thr Ala Met Ile Gly Arg Tyr Trp Asn Asp Thr Ser Ser 435 440 445

Gln Leu Arg Glu Ser Lys Gly Gly Glu Met Leu Thr Ala Leu Asp Phe 450 455 460

His Met Lys Glu Tyr Gly Leu Thr Lys Glu Glu Ala Ala Ser Lys Phe 465 470 475 480

Glu Gly Leu Val Glu Glu Thr Trp Lys Asp Ile Asn Lys Glu Phe Ile 485 490 495

Ala Thr Thr Asn Tyr Asn Val Gly Arg Glu Ile Ala Ile Thr Phe Leu 500 505 510

Asn Tyr Ala Arg Ile Cys Glu Ala Ser Tyr Ser Lys Thr Asp Gly Asp 515 520 525

Ala Tyr Ser Asp Pro Asn Val Ala Lys Ala Asn Val Val Ala Leu Phe 530 535 540

Val Asp Ala Ile Val Phe 545 550 <210> 29 <211> 1653 <212> DNA <213> Mentha piperita <220> <221> CDS <222> (1)..(1653)

Pagina 76

SEQLTXT

<400> 29 atg gct acg aac ggt gtt gtg ata agt tgc cta agg gaa gtt aga ccg 48

Met Ala Thr Asn Gly Val Val Ile Ser Cys Leu Arg Glu Val Arg Pro 1 5 10 15 cct atg acc aaa cac gca cct tca atg tgg acc gat act ttt agc aat 96

Pro Met Thr Lys His Ala Pro Ser Met Trp Thr Asp Thr Phe Ser Asn ttt tcc cta gac gac aag gaa caa caa aaa tgc agt gaa act att gaa 144

Phe Ser Leu Asp Asp Lys Glu Gln Gln Lys Cys Ser Glu Thr Ile Glu gcc ctg aaa cag gaa gca aga ggt atg tta atg gct gcc acc act ccc 192

Ala Leu Lys Gln Glu Ala Arg Gly Met Leu Met Ala Ala Thr Thr Pro 60 cta caa caa atg aca ttg atc gac acg cta gaa aga ctt gga ttg tcc 240

Leu Gln Gln Met Thr Leu Ile Asp Thr Leu Glu Arg Leu Gly Leu Ser 65 70 75 80 ttc cac ttt gaa act gaa att gaa tat aag att gaa ttg ata aac gcc 288

Phe His Phe Glu Thr Glu Ile Glu Tyr Lys Ile Glu Leu Ile Asn Ala 85 90 95 gcc gag gat gat ggc ttt gac ctt ttt gca aca gct ctt aga ttt agg 336

Ala Glu Asp Asp Gly Phe Asp Leu Phe Ala Thr Ala Leu Arg Phe Arg 100 105 110 ttg tta aga cag cat cag aga cac gtc agt tgt gac gtc ttt gat aaa 384

Leu Leu Arg Gln His Gln Arg His Val Ser Cys Asp Val Phe Asp Lys 115 120 125 ttt att gat aaa gat ggt aaa ttc gaa gag agc ctg tca aac aac gtt 432

Phe Ile Asp Lys Asp Gly Lys Phe Glu Glu Ser Leu Ser Asn Asn Val 130 135 140 gaa ggc tta ctg tcg tta tat gaa gct gca cat gtc ggg ttc agg gaa 480

Glu Gly Leu Leu Ser Leu Tyr Glu Ala Ala His Val Gly Phe Arg Glu 145 150 155 160 gaa aga atc tta cag gag gct gta aac ttt act aga cac cat cta gag 528

Glu Arg Ile Leu Gln Glu Ala Val Asn Phe Thr Arg His His Leu Glu 165 170 175 gga gct gaa ttg gat cag tcc ccg ctt ctt att agg gaa aaa gtt aaa 576

Gly Ala Glu Leu Asp Gln Ser Pro Leu Leu Ile Arg Glu Lys Val Lys 180 185 190 aga gct ctt gaa cat cca ttg cac aga gat ttt cca atc gtt tat gca 624

Arg Ala Leu Glu His Pro Leu His Arg Asp Phe Pro Ile Val Tyr Ala 195 200 205

Pagina 77

SEQLTXT cgt ctg ttt att tct att tac gaa aaa gat gac tcc agg gat gaa tta 672

Arg Leu Phe Ile Ser Ile Tyr Glu Lys Asp Asp Ser Arg Asp Glu Leu 210 215 220 ctg cta aaa ttg tcc aaa gtt aac ttt aag ttt atg caa aac ctt tat 720

Leu Leu Lys Leu Ser Lys Val Asn Phe Lys Phe Met Gln Asn Leu Tyr 225 230 235 240 aag gaa gaa tta agt caa ttg agc aga tgg tgg aac act tgg aac ttg 768

Lys Glu Glu Leu Ser Gln Leu Ser Arg Trp Trp Asn Thr Trp Asn Leu 245 250 255 aaa tct aaa ttg cct tac gct aga gat aga gtt gtg gag gct tat gtt 816

Lys Ser Lys Leu Pro Tyr Ala Arg Asp Arg Val Val Glu Ala Tyr Val 260 265 270 tgg ggc gta ggt tat cat tat gaa cct caa tat tct tac gtt aga atg 864

Trp Gly Val Gly Tyr His Tyr Glu Pro Gln Tyr Ser Tyr Val Arg Met 275 280 285 ggc ttg gct aaa ggc gtg ctg atc tgt ggc att atg gat gat aca tat 912

Gly Leu Ala Lys Gly Val Leu Ile Cys Gly Ile Met Asp Asp Thr Tyr 290 295 300 gat aat tat gcc acc ttg aat gag gca cag ctt ttc aca caa gta ctg 960

Asp Asn Tyr Ala Thr Leu Asn Glu Ala Gln Leu Phe Thr Gln Val Leu 305 310 315 320 gat aaa tgg gat agg gat gag gca gaa agg ttg cca gaa tac atg aag 1008

Asp Lys Trp Asp Arg Asp Glu Ala Glu Arg Leu Pro Glu Tyr Met Lys 325 330 335 ata gtt tac aga ttt att ttg agc att tat gag aac tat gaa aga gat 1056

Ile Val Tyr Arg Phe Ile Leu Ser Ile Tyr Glu Asn Tyr Glu Arg Asp 340 345 350 gca gcc aaa ctt ggt aaa tct ttc gcc gcc cca tac ttt aaa gaa acg 1104

Ala Ala Lys Leu Gly Lys Ser Phe Ala Ala Pro Tyr Phe Lys Glu Thr 355 360 365 gta aaa caa ttg gct cgt gcg ttc aac gaa gag caa aag tgg gtt atg 1152

Val Lys Gln Leu Ala Arg Ala Phe Asn Glu Glu Gln Lys Trp Val Met 370 375 380 gaa aga caa ttg cca tca ttt caa gac tac gtg aag aat tca gaa aaa 1200

Glu Arg Gln Leu Pro Ser Phe Gln Asp Tyr Val Lys Asn Ser Glu Lys 385 390 395 400 act tca tgt att tac acc atg ttc gct agt atc att cca gga ttg aaa 1248

Thr Ser Cys Ile Tyr Thr Met Phe Ala Ser Ile Ile Pro Gly Leu Lys 405 410 415

Pagina 78

SEQLTXT tca gta aca caa gaa acg ata gat tgg atc aaa agt gaa ccg acc cta 1296

Ser Val Thr Gln Glu Thr Ile Asp Trp Ile Lys Ser Glu Pro Thr Leu 420 425 430 gct act tct act gca atg ata ggt aga tat tgg aat gac acg tca tcc 1344

Ala Thr Ser Thr Ala Met Ile Gly Arg Tyr Trp Asn Asp Thr Ser Ser 435 440 445 cag cta agg gaa agc aaa ggt ggt gaa atg ttg aca gct tta gat ttt 1392

Gln Leu Arg Glu Ser Lys Gly Gly Glu Met Leu Thr Ala Leu Asp Phe 450 455 460 cac atg aaa gaa tac ggt ctt aca aaa gaa gag gct gct tca aag ttc 14409

His Met Lys Glu Tyr Gly Leu Thr Lys Glu Glu Ala Ala Ser Lys Phe 465 470 475 480 gag ggg cta gtg gag gaa act tgg aaa gat atc aat aag gag ttc att 1488

Glu Gly Leu Val Glu Glu Thr Trp Lys Asp Ile Asn Lys Glu Phe Ile 485 490 495 gct act acc aat tac aac gtt ggt cgt gag ata gct att act ttt ttg 1536

Ala Thr Thr Asn Tyr Asn Val Gly Arg Glu Ile Ala Ile Thr Phe Leu 500 505 510 aac tac gcg aga att tgt gaa gcc tct tat agt aaa acg gac ggt gat 1584

Asn Tyr Ala Arg Ile Cys Glu Ala Ser Tyr Ser Lys Thr Asp Gly Asp 515 520 525 gcc tac tcc gat cca aat gtt gcg aag gct aac gtt gta gca tta ttt 1632

Ala Tyr Ser Asp Pro Asn Val Ala Lys Ala Asn Val Val Ala Leu Phe 530 535 540 gtg gat gcc ata gtg ttc tag 1653

Val Asp Ala Ile Val Phe 545 550 <210> 30 <211> 550 <212> PRT <213> Mentha piperita <400> 30

Met Ala Thr Asn Gly Val Val Ile Ser Cys Leu Arg Glu Val Arg Pro 1 5 10 15

Pro Met Thr Lys His Ala Pro Ser Met Trp Thr Asp Thr Phe Ser Asn

Pagina 79

SEQLTXT

Phe Ser Leu Asp Asp Lys Glu Gln Gln Lys Cys Ser Glu Thr Ile Glu

Ala Leu Lys Gln Glu Ala Arg Gly Met Leu Met Ala Ala Thr Thr Pro 60

Leu Gln Gln Met Thr Leu Ile Asp Thr Leu Glu Arg Leu Gly Leu Ser 65 70 75 80

Phe His Phe Glu Thr Glu Ile Glu Tyr Lys Ile Glu Leu Ile Asn Ala 85 90 95

Ala Glu Asp Asp Gly Phe Asp Leu Phe Ala Thr Ala Leu Arg Phe Arg 100 105 110

Leu Leu Arg Gln His Gln Arg His Val Ser Cys Asp Val Phe Asp Lys 115 120 125

Phe Ile Asp Lys Asp Gly Lys Phe Glu Glu Ser Leu Ser Asn Asn Val 130 135 140

Glu Gly Leu Leu Ser Leu Tyr Glu Ala Ala His Val Gly Phe Arg Glu 145 150 155 160

Glu Arg Ile Leu Gln Glu Ala Val Asn Phe Thr Arg His His Leu Glu 165 170 175

Gly Ala Glu Leu Asp Gln Ser Pro Leu Leu Ile Arg Glu Lys Val Lys 180 185 190

Arg Ala Leu Glu His Pro Leu His Arg Asp Phe Pro Ile Val Tyr Ala 195 200 205

Arg Leu Phe Ile Ser Ile Tyr Glu Lys Asp Asp Ser Arg Asp Glu Leu 210 215 220

Leu Leu Lys Leu Ser Lys Val Asn Phe Lys Phe Met Gln Asn Leu Tyr 225 230 235 240

Pagina 860

SEQLTXT

Lys Glu Glu Leu Ser Gln Leu Ser Arg Trp Trp Asn Thr Trp Asn Leu 245 250 255

Lys Ser Lys Leu Pro Tyr Ala Arg Asp Arg Val Val Glu Ala Tyr Val 260 265 270

Trp Gly Val Gly Tyr His Tyr Glu Pro Gln Tyr Ser Tyr Val Arg Met 275 280 285

Gly Leu Ala Lys Gly Val Leu Ile Cys Gly Ile Met Asp Asp Thr Tyr 290 295 300

Asp Asn Tyr Ala Thr Leu Asn Glu Ala Gln Leu Phe Thr Gln Val Leu 305 310 315 320

Asp Lys Trp Asp Arg Asp Glu Ala Glu Arg Leu Pro Glu Tyr Met Lys 325 330 335

Ile Val Tyr Arg Phe Ile Leu Ser Ile Tyr Glu Asn Tyr Glu Arg Asp 340 345 350

Ala Ala Lys Leu Gly Lys Ser Phe Ala Ala Pro Tyr Phe Lys Glu Thr 355 360 365

Val Lys Gln Leu Ala Arg Ala Phe Asn Glu Glu Gln Lys Trp Val Met 370 375 380

Glu Arg Gln Leu Pro Ser Phe Gln Asp Tyr Val Lys Asn Ser Glu Lys 385 390 395 400

Thr Ser Cys Ile Tyr Thr Met Phe Ala Ser Ile Ile Pro Gly Leu Lys 405 410 415

Ser Val Thr Gln Glu Thr Ile Asp Trp Ile Lys Ser Glu Pro Thr Leu 420 425 430

Ala Thr Ser Thr Ala Met Ile Gly Arg Tyr Trp Asn Asp Thr Ser Ser 435 440 445

Pagina 81

SEQLTXT

Gln Leu Arg Glu Ser Lys Gly Gly Glu Met Leu Thr Ala Leu Asp Phe 450 455 460

His Met Lys Glu Tyr Gly Leu Thr Lys Glu Glu Ala Ala Ser Lys Phe 465 470 475 480

Glu Gly Leu Val Glu Glu Thr Trp Lys Asp Ile Asn Lys Glu Phe Ile 485 490 495

Ala Thr Thr Asn Tyr Asn Val Gly Arg Glu Ile Ala Ile Thr Phe Leu 500 505 510

Asn Tyr Ala Arg Ile Cys Glu Ala Ser Tyr Ser Lys Thr Asp Gly Asp 515 520 525

Ala Tyr Ser Asp Pro Asn Val Ala Lys Ala Asn Val Val Ala Leu Phe 530 535 540

Val Asp Ala Ile Val Phe 545 550 <21e> 31 <211> 1653 <212> DNA <213> Mentha piperita <220> <221> CDS <222> (1)..(1653) <400> 31 atg gcc acc aat ggt gtt gtt att tca tgt ttg cgt gaa gtc aga cca 48

Met Ala Thr Asn Gly Val Val Ile Ser Cys Leu Arg Glu Val Arg Pro 1 5 10 15 cca atg act aag cat gct cca tcc atg tgg act gat acc ttc tct aac 96

Pro Met Thr Lys His Ala Pro Ser Met Trp Thr Asp Thr Phe Ser Asn ttc tcc tta gat gat aag gaa caa caa aaa tgt tca gag act atc gag 144

Phe Ser Leu Asp Asp Lys Glu Gln Gln Lys Cys Ser Glu Thr Ile Glu

Pagina 82

SEQLTXT gct tta aag caa gaa gct aga ggt atg tta atg gcc gct acc acc cca 192

Ala Leu Lys Gln Glu Ala Arg Gly Met Leu Met Ala Ala Thr Thr Pro 60 cta caa caa atg act tta att gac aca ttg gaa cgt cta ggt tta tcc 240

Leu Gln Gln Met Thr Leu Ile Asp Thr Leu Glu Arg Leu Gly Leu Ser 65 70 75 80 ttc cac ttt gaa act gaa att gag tac aaa att gaa ttg atc aat gcc 288

Phe His Phe Glu Thr Glu Ile Glu Tyr Lys Ile Glu Leu Ile Asn Ala 85 90 95 gct gag gac gac ggt ttt gat ttg ttt gcc acc gcc ttg cgt ttt aga 336

Ala Glu Asp Asp Gly Phe Asp Leu Phe Ala Thr Ala Leu Arg Phe Arg 100 105 110 cta cta aga caa cat caa aga cac gtt tcc tgt gat gtt ttt gat aag 384

Leu Leu Arg Gln His Gln Arg His Val Ser Cys Asp Val Phe Asp Lys 115 120 125 ttc atc gat aag gat ggt aag ttt gag gaa tct ttg tct aac aac gtt 432

Phe Ile Asp Lys Asp Gly Lys Phe Glu Glu Ser Leu Ser Asn Asn Val 130 135 140 gag ggt ttg tta tcc ttg tac gaa gca gct cat gtt ggt ttc cgt gag 480

Glu Gly Leu Leu Ser Leu Tyr Glu Ala Ala His Val Gly Phe Arg Glu 145 150 155 160 gaa aga atc tta caa gaa gcc gtc aac ttc act aga cat cat ttg gaa 528

Glu Arg Ile Leu Gln Glu Ala Val Asn Phe Thr Arg His His Leu Glu 165 170 175 ggt gct gaa cta gat caa tcc cca tta tta atc aga gaa aag gtt aag 576

Gly Ala Glu Leu Asp Gln Ser Pro Leu Leu Ile Arg Glu Lys Val Lys 180 185 190 aga gct tta gaa cat cca ttg cat aga gac ttc cca att gtt tat gcc 624

Arg Ala Leu Glu His Pro Leu His Arg Asp Phe Pro Ile Val Tyr Ala 195 200 205 aga ttg ttc atc agt atc tat gaa aag gac gac tct aga gac gaa ttg 672

Arg Leu Phe Ile Ser Ile Tyr Glu Lys Asp Asp Ser Arg Asp Glu Leu 210 215 220 ttg ttg aag ttg tct aag gtt aac ttc aaa ttc atg caa aac tta tac 720

Leu Leu Lys Leu Ser Lys Val Asn Phe Lys Phe Met Gln Asn Leu Tyr 225 230 235 240 aag gaa gaa ttg tcc caa tta tct aga tgg tgg aac acc tgg aac ttg 768

Lys Glu Glu Leu Ser Gln Leu Ser Arg Trp Trp Asn Thr Trp Asn Leu 245 250 255

Pagina 83

SEQLTXT aag tcc aag tta cct tac gcc aga gac aga gtt gtc gaa gct tat gtt 816

Lys Ser Lys Leu Pro Tyr Ala Arg Asp Arg Val Val Glu Ala Tyr Val 260 265 270 tgg ggt gtt ggt tac cat tac gaa cca caa tat tct tat gtc aga atg 864

Trp Gly Val Gly Tyr His Tyr Glu Pro Gln Tyr Ser Tyr Val Arg Met 275 280 285 ggt ttg gct aaa ggt gtc tta att tgt ggt atc atg gac gat acc tac 912

Gly Leu Ala Lys Gly Val Leu Ile Cys Gly Ile Met Asp Asp Thr Tyr 290 295 300 gac aac tac gct acc ttg aac gaa gcc caa cta ttc acc caa gtc tta 960

Asp Asn Tyr Ala Thr Leu Asn Glu Ala Gln Leu Phe Thr Gln Val Leu 305 310 315 320 gac aag tgg gac aga gat gaa gca gaa aga cta cct gaa tac atg aag 1008

Asp Lys Trp Asp Arg Asp Glu Ala Glu Arg Leu Pro Glu Tyr Met Lys 325 330 335 atc gtt tat aga ttc att tta tcc att tac gaa aac tat gaa cgt gat 1056

Ile Val Tyr Arg Phe Ile Leu Ser Ile Tyr Glu Asn Tyr Glu Arg Asp 340 345 350 gct gcc aag ttg ggt aag tcc ttc gcc get cct tac ttc aaa gaa aca 1104

Ala Ala Lys Leu Gly Lys Ser Phe Ala Ala Pro Tyr Phe Lys Glu Thr 355 360 365 gtc aag caa ttg gcc aga gcc ttc aat gaa gaa caa aag tgg gtt atg 1152

Val Lys Gln Leu Ala Arg Ala Phe Asn Glu Glu Gln Lys Trp Val Met 370 375 380 gaa aga caa tta cca tct ttt cag gat tat gtt aag aat tct gaa aag 1200

Glu Arg Gln Leu Pro Ser Phe Gln Asp Tyr Val Lys Asn Ser Glu Lys 385 390 395 400 act tcc tgt atc tac acc atg ttc gct tct att att cct ggc tta aag 1248

Thr Ser Cys Ile Tyr Thr Met Phe Ala Ser Ile Ile Pro Gly Leu Lys 405 410 415 tct gtt act caa gaa act att gac tgg att aag tca gaa cca acc ttg 1296

Ser Val Thr Gln Glu Thr Ile Asp Trp Ile Lys Ser Glu Pro Thr Leu 420 425 430 gcc act agt acc gct atg att ggc cgt tac tgg aac gat acc tct agt 1344

Ala Thr Ser Thr Ala Met Ile Gly Arg Tyr Trp Asn Asp Thr Ser Ser 435 440 445 caa cta aga gaa tct aaa ggt ggt gaa atg ttg acc gcc ttg gac ttt 1392

Gln Leu Arg Glu Ser Lys Gly Gly Glu Met Leu Thr Ala Leu Asp Phe 450 455 460

Pagina 84

SEQLTXT cac atg aaa gaa tat ggt ttg aca aag gaa gag gca gct tct aaa ttc 14409

His Met Lys Glu Tyr Gly Leu Thr Lys Glu Glu Ala Ala Ser Lys Phe 465 470 475 480 gag ggc tta gtc gaa gaa aca tgg aag gac atc aac aag gaa ttc att 1488

Glu Gly Leu Val Glu Glu Thr Trp Lys Asp Ile Asn Lys Glu Phe Ile 485 490 495 gct act act aat tac aac gtc ggt cgt gaa att gct atc acc ttc ttg 1536

Ala Thr Thr Asn Tyr Asn Val Gly Arg Glu Ile Ala Ile Thr Phe Leu 500 505 510 aac tac gct aga atc tgt gaa gct tcc tat tcc aaa acc gac ggt gat 1584

Asn Tyr Ala Arg Ile Cys Glu Ala Ser Tyr Ser Lys Thr Asp Gly Asp 515 520 525 gcc tat tct gat cca aat gtt gcc aaa gct aac gtc gtt get ttg ttt 1632

Ala Tyr Ser Asp Pro Asn Val Ala Lys Ala Asn Val Val Ala Leu Phe 530 535 540 gtc gac gct atc gtt ttc taa 1653

Val Asp Ala Ile Val Phe 545 550 <210> 32 <211> 550 <212> PRT <213> Mentha piperita <400> 32

Met Ala Thr Asn Gly Val Val Ile Ser Cys Leu Arg Glu Val Arg Pro 1 5 10 15

Pro Met Thr Lys His Ala Pro Ser Met Trp Thr Asp Thr Phe Ser Asn

Phe Ser Leu Asp Asp Lys Glu Gln Gln Lys Cys Ser Glu Thr Ile Glu

Ala Leu Lys Gln Glu Ala Arg Gly Met Leu Met Ala Ala Thr Thr Pro 60

Leu Gln Gln Met Thr Leu Ile Asp Thr Leu Glu Arg Leu Gly Leu Ser 65 70 75 80

Pagina 85

SEQLTXT

Phe His Phe Glu Thr Glu Ile Glu Tyr Lys Ile Glu Leu Ile Asn Ala 85 90 95

Ala Glu Asp Asp Gly Phe Asp Leu Phe Ala Thr Ala Leu Arg Phe Arg 100 105 110

Leu Leu Arg Gln His Gln Arg His Val Ser Cys Asp Val Phe Asp Lys 115 120 125

Phe Ile Asp Lys Asp Gly Lys Phe Glu Glu Ser Leu Ser Asn Asn Val 130 135 140

Glu Gly Leu Leu Ser Leu Tyr Glu Ala Ala His Val Gly Phe Arg Glu 145 150 155 160

Glu Arg Ile Leu Gln Glu Ala Val Asn Phe Thr Arg His His Leu Glu 165 170 175

Gly Ala Glu Leu Asp Gln Ser Pro Leu Leu Ile Arg Glu Lys Val Lys 180 185 190

Arg Ala Leu Glu His Pro Leu His Arg Asp Phe Pro Ile Val Tyr Ala 195 200 205

Arg Leu Phe Ile Ser Ile Tyr Glu Lys Asp Asp Ser Arg Asp Glu Leu 210 215 220

Leu Leu Lys Leu Ser Lys Val Asn Phe Lys Phe Met Gln Asn Leu Tyr 225 230 235 240

Lys Glu Glu Leu Ser Gln Leu Ser Arg Trp Trp Asn Thr Trp Asn Leu 245 250 255

Lys Ser Lys Leu Pro Tyr Ala Arg Asp Arg Val Val Glu Ala Tyr Val 260 265 270

Trp Gly Val Gly Tyr His Tyr Glu Pro Gln Tyr Ser Tyr Val Arg Met 275 280 285

Pagina 86

SEQLTXT

Gly Leu Ala Lys Gly Val Leu Ile Cys Gly Ile Met Asp Asp Thr Tyr 290 295 300

Asp Asn Tyr Ala Thr Leu Asn Glu Ala Gln Leu Phe Thr Gln Val Leu 305 310 315 320

Asp Lys Trp Asp Arg Asp Glu Ala Glu Arg Leu Pro Glu Tyr Met Lys 325 330 335

Ile Val Tyr Arg Phe Ile Leu Ser Ile Tyr Glu Asn Tyr Glu Arg Asp 340 345 350

Ala Ala Lys Leu Gly Lys Ser Phe Ala Ala Pro Tyr Phe Lys Glu Thr 355 360 365

Val Lys Gln Leu Ala Arg Ala Phe Asn Glu Glu Gln Lys Trp Val Met 370 375 380

Glu Arg Gln Leu Pro Ser Phe Gln Asp Tyr Val Lys Asn Ser Glu Lys 385 390 395 400

Thr Ser Cys Ile Tyr Thr Met Phe Ala Ser Ile Ile Pro Gly Leu Lys 405 410 415

Ser Val Thr Gln Glu Thr Ile Asp Trp Ile Lys Ser Glu Pro Thr Leu 420 425 430

Ala Thr Ser Thr Ala Met Ile Gly Arg Tyr Trp Asn Asp Thr Ser Ser 435 440 445

Gln Leu Arg Glu Ser Lys Gly Gly Glu Met Leu Thr Ala Leu Asp Phe 450 455 460

His Met Lys Glu Tyr Gly Leu Thr Lys Glu Glu Ala Ala Ser Lys Phe 465 470 475 480

Glu Gly Leu Val Glu Glu Thr Trp Lys Asp Ile Asn Lys Glu Phe Ile 485 490 495

Pagina 87

SEQLTXT

Ala Thr Thr Asn Tyr Asn Val Gly Arg Glu Ile Ala Ile Thr Phe Leu 500 505 510

Asn Tyr Ala Arg Ile Cys Glu Ala Ser Tyr Ser Lys Thr Asp Gly Asp 515 520 525

Ala Tyr Ser Asp Pro Asn Val Ala Lys Ala Asn Val Val Ala Leu Phe 530 535 540

Val Asp Ala Ile Val Phe 545 550 <210> 33 <211> 1614 <212> DNA <213> Salvia miltiorrhiza <220> <221> CDS <222> (1)..(1614) <400> 33 atg gaa gaa gtt aga aga tct gct aac tta ggt gcc agc ata tgg gat 48

Met Glu Glu Val Arg Arg Ser Ala Asn Leu Gly Ala Ser Ile Trp Asp 1 5 10 15 gac gat tac att cag tcc ttg gcc tca ttg tac aca ggg gaa aaa tac 96

Asp Asp Tyr Ile Gln Ser Leu Ala Ser Leu Tyr Thr Gly Glu Lys Tyr gtt gca gag gct gag aaa ttg aaa aat ttg gtc aag ata atg ata gat 144

Val Ala Glu Ala Glu Lys Leu Lys Asn Leu Val Lys Ile Met Ile Asp gaa act gaa gac gaa ttg gac caa tta gaa ttg att gat tct ctt caa 192

Glu Thr Glu Asp Glu Leu Asp Gln Leu Glu Leu Ile Asp Ser Leu Gln 60 cgt ctt ggc ttg tgc aac cat ttc gaa gac aga atc gcc aag aaa tta 240

Arg Leu Gly Leu Cys Asn His Phe Glu Asp Arg Ile Ala Lys Lys Leu 65 70 75 80 gac agc att tat gaa gcc gag aag tgc atg gaa gaa gat ctt cac gta 288

Asp Ser Ile Tyr Glu Ala Glu Lys Cys Met Glu Glu Asp Leu His Val 85 90 95 acc gcc ttg aaa ttt aga ttg tta cgt caa cac ggt tat cat gtg ccg 336

Pagina 88

SEQLTXT

Thr Ala Leu Lys Phe Arg Leu Leu Arg Gln His Gly Tyr His Val Pro 100 105 110 cag gaa gtg ttc tgt tca ttc atg gat gac gtt ggt aat ttt aag gca 384

Gln Glu Val Phe Cys Ser Phe Met Asp Asp Val Gly Asn Phe Lys Ala 115 120 125 tcg ttg tgt gag gat gtc aga ggt gtt gtg agt ttg tac gaa gca tct 432

Ser Leu Cys Glu Asp Val Arg Gly Val Val Ser Leu Tyr Glu Ala Ser 130 135 140 tac tta tct atg gaa ggt gaa tcc atc tta gac ctt gct aag gat ttc 480

Tyr Leu Ser Met Glu Gly Glu Ser Ile Leu Asp Leu Ala Lys Asp Phe 145 150 155 160 tca ctt aat cat tta act aga aga atg gat aaa att aca gaa cca aga 528

Ser Leu Asn His Leu Thr Arg Arg Met Asp Lys Ile Thr Glu Pro Arg 165 170 175 ttg gcc gag caa gtt cgt cac gcc cta gaa gtt cca tta cat tgg aga 576

Leu Ala Glu Gln Val Arg His Ala Leu Glu Val Pro Leu His Trp Arg 180 185 190 gtc caa aga ctg gaa gct agg tgg tac att aag gct tat gag tca agg 624

Val Gln Arg Leu Glu Ala Arg Trp Tyr Ile Lys Ala Tyr Glu Ser Arg 195 200 205 tcg tcc gct aac ttg att ctg gta gaa ttg gct aaa ttg gat ttt aat 672

Ser Ser Ala Asn Leu Ile Leu Val Glu Leu Ala Lys Leu Asp Phe Asn 210 215 220 atg gtg caa gct aca tac caa caa gag tta aaa aga atg tct aga tgg 720

Met Val Gln Ala Thr Tyr Gln Gln Glu Leu Lys Arg Met Ser Arg Trp 225 230 235 240 tat aag gaa acc ggt ttg aca gaa aag tta ggt ttc gca aga cac aga 768

Tyr Lys Glu Thr Gly Leu Thr Glu Lys Leu Gly Phe Ala Arg His Arg 245 250 255 tta gca gaa tgc ttt ttg tgg gcc atg ggg ttt gtt cct gaa cca caa 816

Leu Ala Glu Cys Phe Leu Trp Ala Met Gly Phe Val Pro Glu Pro Gln 260 265 270 ttg ggt tac tcg agg gag att ctt att aag atc gcc gta ttt att acg 864

Leu Gly Tyr Ser Arg Glu Ile Leu Ile Lys Ile Ala Val Phe Ile Thr 275 280 285 atc att gac gac att tac gat gtt tac ggg aca ttg gaa gaa cta cag 912

Ile Ile Asp Asp Ile Tyr Asp Val Tyr Gly Thr Leu Glu Glu Leu Gln 290 295 300 ttg ttc acc aac act ata gaa cgt tgg gac atc aac agc cta gac aac 960

Pagina 89

SEQLTXT

Leu Phe Thr Asn Thr Ile Glu Arg Trp Asp Ile Asn Ser Leu Asp Asn 305 310 315 320 ttg cca gaa tat atg aga atc tgt ttt ctt gca tta ttt aat tct gca 1008

Leu Pro Glu Tyr Met Arg Ile Cys Phe Leu Ala Leu Phe Asn Ser Ala 325 330 335 aat gaa cta gct tac cac att tta aga gat cag ggt ttt aac gtt atc 1056

Asn Glu Leu Ala Tyr His Ile Leu Arg Asp Gln Gly Phe Asn Val Ile 340 345 350 tca aat cta agg aaa tta tgg gcc gaa ttg tgc cgt gca tac tat cta 1104

Ser Asn Leu Arg Lys Leu Trp Ala Glu Leu Cys Arg Ala Tyr Tyr Leu 355 360 365 gaa gcc aga tgg ttt cat tcc ggt tat gtg ccc aca acc gat gaa tat 1152

Glu Ala Arg Trp Phe His Ser Gly Tyr Val Pro Thr Thr Asp Glu Tyr 370 375 380 ttg aat act gct tgg aag tca ata tca gga cca tta tta tta ttt tat 1200

Leu Asn Thr Ala Trp Lys Ser Ile Ser Gly Pro Leu Leu Leu Phe Tyr 385 390 395 400 ggc tat ttt agt act aat cct att aac aag aaa gaa tta cag tct tta 1248

Gly Tyr Phe Ser Thr Asn Pro Ile Asn Lys Lys Glu Leu Gln Ser Leu 405 410 415 gag caa tat cca ggg att att aga tgg cca agc acc gtt tta aga tta 1296

Glu Gln Tyr Pro Gly Ile Ile Arg Trp Pro Ser Thr Val Leu Arg Leu 420 425 430 gct gat gat tta ggt acc tcg tct gag gaa atg aag agg ggt gac gtt 1344

Ala Asp Asp Leu Gly Thr Ser Ser Glu Glu Met Lys Arg Gly Asp Val 435 440 445 cca aag tcc gtc caa tgc tat atg aca gag aca ggt tgt tcg gaa gaa 1392

Pro Lys Ser Val Gln Cys Tyr Met Thr Glu Thr Gly Cys Ser Glu Glu 450 455 460 gac gca agg aag cat att aaa cat cta ata gaa aca gct tta aaa aga 14409

Asp Ala Arg Lys His Ile Lys His Leu Ile Glu Thr Ala Leu Lys Arg 465 470 475 480 atg aat aaa gag ata ctg atg gag aaa ccg cta aag aat ttt gga cac 1488

Met Asn Lys Glu Ile Leu Met Glu Lys Pro Leu Lys Asn Phe Gly His 485 490 495 ata gcc atg aat tta ggt aga ata agt ctg tgt atg tac caa cat ggg 1536

Ile Ala Met Asn Leu Gly Arg Ile Ser Leu Cys Met Tyr Gln His Gly 500 505 510 gat ggg ttt ggc ttg cct cat tca gag aca aag agg aac ttg gta agc 1584

Pagina 90

SEQLTXT

Asp Gly Phe Gly Leu Pro His Ser Glu Thr Lys Arg Asn Leu Val Ser 515 520 525 ttg tta gtt cag cct ttt cct atg ccg tga 1614

Leu Leu Val Gln Pro Phe Pro Met Pro 530 535 <210> 34 <211> 537 <212> PRT <213> Salvia miltiorrhiza <400> 34

Met Glu Glu Val Arg Arg Ser Ala Asn Leu Gly Ala Ser Ile Trp Asp 1 5 10 15

Asp Asp Tyr Ile Gln Ser Leu Ala Ser Leu Tyr Thr Gly Glu Lys Tyr

Val Ala Glu Ala Glu Lys Leu Lys Asn Leu Val Lys Ile Met Ile Asp

Glu Thr Glu Asp Glu Leu Asp Gln Leu Glu Leu Ile Asp Ser Leu Gln 60

Arg Leu Gly Leu Cys Asn His Phe Glu Asp Arg Ile Ala Lys Lys Leu 65 70 75 80

Asp Ser Ile Tyr Glu Ala Glu Lys Cys Met Glu Glu Asp Leu His Val 85 90 95

Thr Ala Leu Lys Phe Arg Leu Leu Arg Gln His Gly Tyr His Val Pro 100 105 119

Gln Glu Val Phe Cys Ser Phe Met Asp Asp Val Gly Asn Phe Lys Ala 115 120 125

Ser Leu Cys Glu Asp Val Arg Gly Val Val Ser Leu Tyr Glu Ala Ser 130 135 140

Tyr Leu Ser Met Glu Gly Glu Ser Ile Leu Asp Leu Ala Lys Asp Phe

Pagina 91

SEQLTXT

145 150 155 160

Ser Leu Asn His Leu Thr Arg Arg Met Asp Lys Ile Thr Glu Pro Arg 165 170 175

Leu Ala Glu Gln Val Arg His Ala Leu Glu Val Pro Leu His Trp Arg 180 185 190

Val Gln Arg Leu Glu Ala Arg Trp Tyr Ile Lys Ala Tyr Glu Ser Arg 195 200 205

Ser Ser Ala Asn Leu Ile Leu Val Glu Leu Ala Lys Leu Asp Phe Asn 210 215 220

Met Val Gln Ala Thr Tyr Gln Gln Glu Leu Lys Arg Met Ser Arg Trp 225 230 235 240

Tyr Lys Glu Thr Gly Leu Thr Glu Lys Leu Gly Phe Ala Arg His Arg 245 250 255

Leu Ala Glu Cys Phe Leu Trp Ala Met Gly Phe Val Pro Glu Pro Gln 260 265 270

Leu Gly Tyr Ser Arg Glu Ile Leu Ile Lys Ile Ala Val Phe Ile Thr 275 280 285

Ile Ile Asp Asp Ile Tyr Asp Val Tyr Gly Thr Leu Glu Glu Leu Gln 290 295 300

Leu Phe Thr Asn Thr Ile Glu Arg Trp Asp Ile Asn Ser Leu Asp Asn 305 310 315 320

Leu Pro Glu Tyr Met Arg Ile Cys Phe Leu Ala Leu Phe Asn Ser Ala 325 330 335

Asn Glu Leu Ala Tyr His Ile Leu Arg Asp Gln Gly Phe Asn Val Ile 340 345 350

Ser Asn Leu Arg Lys Leu Trp Ala Glu Leu Cys Arg Ala Tyr Tyr Leu

Pagina 92

SEQLTXT

355 360 365

Glu Ala Arg Trp Phe His Ser Gly Tyr Val Pro Thr Thr Asp Glu Tyr 370 375 380

Leu Asn Thr Ala Trp Lys Ser Ile Ser Gly Pro Leu Leu Leu Phe Tyr 385 390 395 400

Gly Tyr Phe Ser Thr Asn Pro Ile Asn Lys Lys Glu Leu Gln Ser Leu 405 410 415

Glu Gln Tyr Pro Gly Ile Ile Arg Trp Pro Ser Thr Val Leu Arg Leu 420 425 430

Ala Asp Asp Leu Gly Thr Ser Ser Glu Glu Met Lys Arg Gly Asp Val 435 440 445

Pro Lys Ser Val Gln Cys Tyr Met Thr Glu Thr Gly Cys Ser Glu Glu 450 455 460

Asp Ala Arg Lys His Ile Lys His Leu Ile Glu Thr Ala Leu Lys Arg 465 470 475 480

Met Asn Lys Glu Ile Leu Met Glu Lys Pro Leu Lys Asn Phe Gly His 485 490 495

Ile Ala Met Asn Leu Gly Arg Ile Ser Leu Cys Met Tyr Gln His Gly 500 505 510

Asp Gly Phe Gly Leu Pro His Ser Glu Thr Lys Arg Asn Leu Val Ser 515 520 525

Leu Leu Val Gln Pro Phe Pro Met Pro 530 535 <210> 35 <211> 1602 <212> DNA <213> Zea diploperennis

Pagina 93

SEQLTXT

<220> <221> CDS <222> (1)..(1602) <400> 35 atg gat gcg act gcg ttt cat cca tca ttg tgg ggc gac ttt ttc gtg 48

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15 aag tac aag cct cct acg gcc cca aag agg ggt cat atg act caa aga 96

Lys Tyr Lys Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Gln Arg gct gaa tta ctt aag gaa gag gtc aga aag acc ctt aaa gCg gca gcg 144

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala aat caa att aaa aat gct ctg gat ttg ata att act ttg caa cgt tta 192

Asn Gln Ile Lys Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60 gga cta gac cat cat tac gag aat gaa att tct gaa ttg ctg aga ttc 240

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80 gtt tac tca agt tca gat tat gat gat aaa gat ttg tat gtg gta tcc 288

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95 ttg agg ttc tac cta ttg agg aaa cat ggc cat tgc gtc tcc agt gac 336

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Cys Val Ser Ser Asp 100 105 110 gtc ttc acc tct ttt aaa gac gaa gag ggg aac ttt gta gtc gac gat 384

Val Phe Thr Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125 acc aag tgc tta ctt tcc ctt tac aat gca gca tat cta aga aca cat 432

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Leu Arg Thr His 130 135 140 gga gaa aag gta ctg gat gaa gca ata aca ttt acc aga aga cag ctt 480

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160 gaa gca tta tta ttg gat agt tta gag cca gct ttg gct gat gaa gtt 528

Glu Ala Leu Leu Leu Asp Ser Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175 cat ctg act cta caa acc cct ttg ttt aga aga ctt agg atc ttg gaa 576

His Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu

Pagina 94

SEQLTXT

180 185 190 gct gtg aat tat atc cca att tat ggc aag gaa gcg ggc cgt aac gag 624

Ala Val Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205 gcc att ttg gag ttg gca aag tta aat ttt aat ctt gcg caa cta att 672

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220 tac tgt gaa gaa ctg aaa gaa ata act ctt tgg tgg aag caa cta aac 720

Tyr Cys Glu Glu Leu Lys Glu Ile Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240 gta gag act aat ctg tcg ttc atc aga gac aga atc gta gaa tgc cat 768

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255 ttt tgg atg act ggc gca tgc tgc gaa cct caa tac tcc ctt tct cgt 816

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Gln Tyr Ser Leu Ser Arg 260 265 270 gtc ata gcc acc aaa atg acc gct ctt ata acc gtc tta gat gat atg 864

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285 atg gac act tat tcc acc aca gaa gaa gct atg ttg tta gcc gaa gca 912

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300 att tat agg tgg gaa gag agt gct gct gaa tta ttg cca ggt tac atg 960

Ile Tyr Arg Trp Glu Glu Ser Ala Ala Glu Leu Leu Pro Gly Tyr Met 305 310 315 320 aag gat ttt tat cta tat ttg cta aag acc att gac tcg tgt ggt gac 1008

Lys Asp Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Asp 325 330 335 gag ctt ggc cca aat aga tct ttc agg acg ttc tac ttg aag gaa atg 1056

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350 tta aag gtt ttt gta cgt ggt agt tcc cag gaa att aag tgg agg aac 1104

Leu Lys Val Phe Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365 gaa aat tac gtt cct aaa acc atc tcg gaa cat cta gaa cat tca ggc 1152

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380 cca aca gtt ggt gct ttt cag gtt gca tgt tca tct ttt gtt ggt atg 1200

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met

Pagina 95

SEQLTXT

385 390 395 400 gga gac aac att acc aag gaa tca ttc gaa tgg tta cta aca tac ccg 1248

Gly Asp Asn Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415 gag ttg gta aaa agt cta atg aac att gcg agg ttg ttg aat gat act 1296

Glu Leu Val Lys Ser Leu Met Asn Ile Ala Arg Leu Leu Asn Asp Thr 420 425 430 gcc tcc aca aaa aga gaa cag aat gcc ggc cat cac gtt agc act gta 1344

Ala Ser Thr Lys Arg Glu Gln Asn Ala Gly His His Val Ser Thr Val 435 440 445 caa tgt tat atg ctg aaa cac gga act acc atg gat gaa gct tgt gac 1392

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Asp 450 455 460 aag ata aag gag ctt acc gaa gat tcc tgg aaa gat atg atg gaa tta 14409

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480 tac tta acc ccg act gaa cat ccg aaa cta ata gcc cag acc atc gta 1488

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Ile Ala Gln Thr Ile Val 485 490 495 gat ttt gca aga act gct gat tat atg tat aaa gaa acg gac gga ttc 1536

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510 act ttt agc cat act att aaa gat atg att gct aag ttg ttt gtg gac 1584

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525 cca att tca ttg ttt taa 1602

Pro Ile Ser Leu Phe 530 <210> 36 <211> 533 <212> PRT <213> Zea diploperennis <400> 36

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15

Lys Tyr Lys Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Gln Arg

Pagina 96

SEQLTXT

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala

Asn Gln Ile Lys Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Cys Val Ser Ser Asp 100 105 110

Val Phe Thr Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Leu Arg Thr His 130 135 140

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160

Glu Ala Leu Leu Leu Asp Ser Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175

His Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190

Ala Val Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220

Tyr Cys Glu Glu Leu Lys Glu Ile Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240

Pagina 97

SEQLTXT

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Gln Tyr Ser Leu Ser Arg 260 265 270

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300

Ile Tyr Arg Trp Glu Glu Ser Ala Ala Glu Leu Leu Pro Gly Tyr Met 305 310 315 320

Lys Asp Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Asp 325 330 335

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350

Leu Lys Val Phe Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400

Gly Asp Asn Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415

Glu Leu Val Lys Ser Leu Met Asn Ile Ala Arg Leu Leu Asn Asp Thr 420 425 430

Ala Ser Thr Lys Arg Glu Gln Asn Ala Gly His His Val Ser Thr Val 435 440 445

Pagina 98

SEQLTXT

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Asp 450 455 460

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Ile Ala Gln Thr Ile Val 485 490 495

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525

Pro Ile Ser Leu Phe 530 <21e> 37 <211> 1602 <212> DNA <213> Zea mays <220> <221> CDS <222> (1)..(1602) <400> 37 atg gat gct aca gcc ttc cat cct agc tta tgg ggt gat ttc ttt gtg 48

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15 aaa tat aaa cca ccc aca gct cct aaa aga ggt cat atg acg gaa aga 96

Lys Tyr Lys Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Glu Arg gct gaa tta tta aaa gaa gaa gtt cgt aag aca tta aaa gca gcc gct 144

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala aat caa att act aat gct tta gac tta ata atc acc ctt cag agg ctg 192

Asn Gln Ile Thr Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60

Pagina 99

SEQLTXT ggt cta gac cat cat tat gag aac gag att tcg gag cta tta cgt ttc 240

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80 gtc tat tcg tct tct gat tat gac gac aaa gac cta tat gta gtt tct 288

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95 ttg cgt ttt tat ttg ttg aga aaa cat ggt cac tgc gtc tca tca gac 336

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Cys Val Ser Ser Asp 100 105 110 gtt ttc act tca ttt aaa gat gag gag ggt aat ttt gtt gta gat gat 384

Val Phe Thr Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125 act aag tgt ttg cta tcg tta tac aac gca gca tat gta cgt aca cat 432

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Val Arg Thr His 130 135 140 ggt gaa aaa gtt tta gac gaa gcg atc aca ttt aca agg agg caa ttg 480

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160 gaa gct tct ctt ttg gat ccc tta gaa ccc gca tta gcc gac gag gta 528

Glu Ala Ser Leu Leu Asp Pro Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175 cat tta aca ctt caa acc cca cta ttt aga cgt tta aga att ttg gaa 576

His Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190 gca ata aac tat att cca att tac ggc aag gaa gct ggc aga aat gaa 624

Ala Ile Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205 gct ata ttg gag tta gca aag ctt aac ttt aat cta gcg caa ctg ata 672

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220 tat tgt gaa gag ctg aaa gag gtg act cta tgg tgg aaa caa ctt aac 720

Tyr Cys Glu Glu Leu Lys Glu Val Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240 gtt gaa act aat tta tca ttt att aga gac aga att gtg gaa tgt cac 768

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255 ttt tgg atg acg gga gca tgt tgt gag ccg caa tac tct ttg tca cgt 816

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Gln Tyr Ser Leu Ser Arg 260 265 270

Pagina 100

SEQLTXT gtt att gct acg aaa atg act gcg ctt att act gtg ctg gat gac atg 864

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285 atg gac act tac tcg aca acc gaa gaa gct atg ttg tta gcc gaa gca 912

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300 att tat aga tgg gaa gaa aac gct gcg gaa tta ctt ccc aga tat atg 960

Ile Tyr Arg Trp Glu Glu Asn Ala Ala Glu Leu Leu Pro Arg Tyr Met 305 310 315 320 aaa gat ttt tac ctg tac ttg ttg aaa acg att gac tcg tgt ggc gac 1008

Lys Asp Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Asp 325 330 335 gag ttg ggt cct aac agg tcc ttc aga act ttt tat ttg aaa gaa atg 1056

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350 tta aag gtt ttg gtt aga ggc tca tca caa gag att aaa tgg cgt aat 1104

Leu Lys Val Leu Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365 gaa aat tat gta cct aaa act ata tcc gaa cac tta gaa cat tcc ggc 1152

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380 cct act gtt ggg gct ttc cag gta gct tgt agc tct ttc gtt ggt atg 1200

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400 ggg gat tca att acc aaa gaa tca ttt gaa tgg ttg ctg acc tac cca 1248

Gly Asp Ser Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415 gag ttg gct aag tca ctg atg aat atc agt cgt cta cta aat gat act 1296

Glu Leu Ala Lys Ser Leu Met Asn Ile Ser Arg Leu Leu Asn Asp Thr 420 425 430 gcc agt aca aaa agg gaa caa aac gca ggt caa cat gtt tcg acc gta 1344

Ala Ser Thr Lys Arg Glu Gln Asn Ala Gly Gln His Val Ser Thr Val 435 440 445 cag tgc tac atg ctt aaa cac ggt acg acc atg gac gag gca tgt gag 1392

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Glu 450 455 460 aag atc aaa gaa cta act gaa gat tct tgg aag gac atg atg gag tta 14409

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480

Pagina 101

SEQLTXT tat ttg act cca aca gag cat cct aaa ttg att gcc cag acg atc gtg 1488

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Ile Ala Gln Thr Ile Val 485 490 495 gat ttt gca cgt acc gcg gat tat atg tat aaa gaa act gac ggt ttt 1536

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510 acg ttc tct cat aca ata aaa gat atg ata gct aag tta ttt gta gac 1584

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525 ccc att agc cta ttc taa 1602

Pro Ile Ser Leu Phe 530 <210> 38 <211> 533 <212> PRT <213> Zea mays <400> 38

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15

Lys Tyr Lys Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Glu Arg

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala

Asn Gln Ile Thr Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Cys Val Ser Ser Asp 100 105 119

Pagina 102

SEQLTXT

Val Phe Thr Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Val Arg Thr His 130 135 140

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160

Glu Ala Ser Leu Leu Asp Pro Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175

His Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190

Ala Ile Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220

Tyr Cys Glu Glu Leu Lys Glu Val Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Gln Tyr Ser Leu Ser Arg 260 265 270

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300

Ile Tyr Arg Trp Glu Glu Asn Ala Ala Glu Leu Leu Pro Arg Tyr Met 305 310 315 320

Pagina 103

SEQLTXT

Lys Asp Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Asp 325 330 335

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350

Leu Lys Val Leu Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400

Gly Asp Ser Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415

Glu Leu Ala Lys Ser Leu Met Asn Ile Ser Arg Leu Leu Asn Asp Thr 420 425 430

Ala Ser Thr Lys Arg Glu Gln Asn Ala Gly Gln His Val Ser Thr Val 435 440 445

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Glu 450 455 460

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Ile Ala Gln Thr Ile Val 485 490 495

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525

Pagina 104

SEQLTXT

Pro Ile Ser Leu Phe 530 <210> 39 <211> 1602 <212> DNA <213> Zea mays <220> <221> CDS <222> (1)..(1602) <400> 39 atg gac gct act gct ttc cac cca tct ttg tgg ggt gac ttt ttc gtc 48

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15 aag tac aag cca cca act gct cca aaa cgt ggt cac atg act gaa cgt 96

Lys Tyr Lys Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Glu Arg gct gaa ttg ttg aag gaa gaa gtt aga aag act ttg aaa gct gct gca 144

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala aac caa att act aac gct ttg gac cta att atc acc ttg cag aga cta 192

Asn Gln Ile Thr Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60 ggt ttg gac cac cac tac gaa aac gag atc tcc gaa ttg ttg aga ttt 240

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80 gtt tac tcc tcc tcc gat tat gat gat aag gac cta tac gtt gtc tct 288

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95 tta aga ttc tac ttg cta aga aag cac ggt cat tgt gtt tct tct gat 336

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Cys Val Ser Ser Asp 100 105 110 gtc ttt acc tcc ttt aag gat gaa gaa ggt aac ttc gtc gtt gat gat 384

Val Phe Thr Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125 aca aag tgc ttg tta agt ttg tac aac gcc gcc tat gtc aga act cac 432

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Val Arg Thr His 130 135 140

Pagina 105

SEQLTXT ggt gaa aag gtt ttg gat gaa gct att acc ttc act aga aga caa ttg 480

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160 gag gct tca cta ttg gac cca tta gaa cca gct tta gct gac gaa gtt 528

Glu Ala Ser Leu Leu Asp Pro Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175 cac ttg act ttg cag acc cca ttg ttc cgt aga ttg aga att ttg gaa 576

His Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190 gct att aac tac atc cct atc tat ggt aag gaa gct ggt aga aac gag 624

Ala Ile Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205 gca att ttg gaa ttg gct aag tta aac ttc aac ttg gct caa ttg atc 672

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220 tat tgt gaa gaa ttg aaa gaa gtc acc tta tgg tgg aaa caa ttg aac 720

Tyr Cys Glu Glu Leu Lys Glu Val Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240 gtt gaa acc aac ttg tct ttc atc aga gat aga atc gtt gaa tgt cac 768

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255 ttc tgg atg acc ggt gct tgt tgt gaa cca cag tac tcc ttg tct aga 816

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Gln Tyr Ser Leu Ser Arg 260 265 270 gtt att gct acc aag atg acc gca ttg att act gtt ttg gat gac atg 864

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285 atg gat act tat tct acc acc gag gaa gct atg tta ttg gca gaa gca 912

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300 att tat aga tgg gaa gaa aat gcc gcc gaa tta ttg cca cgt tat atg 960

Ile Tyr Arg Trp Glu Glu Asn Ala Ala Glu Leu Leu Pro Arg Tyr Met 305 310 315 320 aag gat ttt tac tta tac tta ttg aag acc atc gac tcc tgt ggt gac 1008

Lys Asp Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Asp 325 330 335 gaa ttg ggt cca aac aga tcc ttt aga act ttc tac cta aag gaa atg 1056

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350

Pagina 106

SEQLTXT ttg aaa gtc cta gtt aga ggt tct tca caa gaa atc aag tgg aga aat 1104

Leu Lys Val Leu Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365 gag aac tac gtt cct aag acc atc agt gaa cat tta gaa cac tca ggt 1152

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380 cct acc gtt ggt gct ttc caa gtt gct tgt tcc tcc ttt gtt ggt atg 1200

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400 ggc gat tct att acc aag gaa agt ttc gag tgg cta ttg acc tat cca 1248

Gly Asp Ser Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415 gaa ttg gct aag tct tta atg aac att tca aga ttg ttg aac gat acc 1296

Glu Leu Ala Lys Ser Leu Met Asn Ile Ser Arg Leu Leu Asn Asp Thr 420 425 430 gcc tct act aaa cgt gaa caa aac gcc ggt caa cac gtt tct act gtt 1344

Ala Ser Thr Lys Arg Glu Gln Asn Ala Gly Gln His Val Ser Thr Val 435 440 445 caa tgt tac atg tta aaa cat ggt aca acc atg gac gaa gcc tgt gaa 1392

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Glu 450 455 460 aag atc aaa gaa tta aca gag gat tct tgg aag gac atg atg gaa cta 14409

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480 tac ttg act cca act gaa cac cca aag ttg att gct caa acc atc gtc 1488

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Ile Ala Gln Thr Ile Val 485 490 495 gac ttc gcc aga acc gcc gat tac atg tat aaa gaa aca gac ggt ttt 1536

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510 act ttt tcc cac acc att aag gac atg atc gct aag ttg ttc gtt gat 1584

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525 cca att tcc tta ttt taa 1602

Pro Ile Ser Leu Phe 530 <210> 40 <211> 533 <212> PRT

Pagina 107

SEQLTXT

<213> Zea mays <400> 40

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15

Lys Tyr Lys Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Glu Arg

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala

Asn Gln Ile Thr Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Cys Val Ser Ser Asp 100 105 119

Val Phe Thr Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Val Arg Thr His 130 135 140

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160

Glu Ala Ser Leu Leu Asp Pro Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175

His Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190

Pagina 108

SEQLTXT

Ala Ile Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220

Tyr Cys Glu Glu Leu Lys Glu Val Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Gln Tyr Ser Leu Ser Arg 260 265 270

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300

Ile Tyr Arg Trp Glu Glu Asn Ala Ala Glu Leu Leu Pro Arg Tyr Met 305 310 315 320

Lys Asp Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Asp 325 330 335

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350

Leu Lys Val Leu Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400

Pagina 109

SEQLTXT

Gly Asp Ser Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415

Glu Leu Ala Lys Ser Leu Met Asn Ile Ser Arg Leu Leu Asn Asp Thr 420 425 430

Ala Ser Thr Lys Arg Glu Gln Asn Ala Gly Gln His Val Ser Thr Val 435 440 445

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Glu 450 455 460

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Ile Ala Gln Thr Ile Val 485 490 495

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525

Pro Ile Ser Leu Phe 530 <210> 41 <211> 1602 <212> DNA <213> Zea perennis <220> <221> CDS <222> (1)..(1602) <400> 41 atg gat gct act gct ttc cat ccg tct ttg tgg ggt gac ttc ttc gtc 48

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15 aaa tac gaa ccg cct act gct cca aaa aga ggt cat atg acg caa agg 96

Pagina 110

SEQLTXT

Lys Tyr Glu Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Gln Arg gca gag cta tta aaa gag gaa gtc aga aaa act ttg aaa gct gcc gct 144

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala aac caa att aag aat gca tta gat ttg att att act ctt caa aga ctt 192

Asn Gln Ile Lys Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60 ggt ttg gac cac cac tat gaa aat gag ata agc gaa ctt ttg cgt ttt 240

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe 65 70 75 80 gtt tat tca agt tct gat tac gat gat aaa gac ttg tac gtt gta tca 288

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95 ctt cgt ttc tat tta tta agg aag cac ggc cac cgt gtc tct tca gac 336

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Arg Val Ser Ser Asp 100 105 110 gta ttc atg tcg ttt aag gat gaa gag ggc aac ttt gtg gtc gac gat 384

Val Phe Met Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125 act aag tgc ctg cta tca tta tac aat gcc gca tat ttg atg act cat 432

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Leu Met Thr His 130 135 140 ggg gaa aaa gtc ctt gat gaa gct ata act ttc acc aga aga cag ttg 480

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160 gaa gct ttg ctg cta gat cct ctg gaa cct gcc ttg gcg gat gaa gtg 528

Glu Ala Leu Leu Leu Asp Pro Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175 tat tta acg tta caa aca cca tta ttt aga cgt tta agg att tta gaa 576

Tyr Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190 gct gtt aat tac atc cca ata tac gga aaa gaa gca gga agg aac gaa 624

Ala Val Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205 gct att tta gaa ttg gct aaa ctg aac ttc aat ttg gca caa ctt atc 672

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220 tat tgt gaa gaa ttg aag gaa gtt acg ttg tgg tgg aaa caa ttg aat 720

Pagina 111

SEQLTXT

Tyr Cys Glu Glu Leu Lys Glu Val Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240 gtg gaa act aat tta agt ttt att aga gat aga atc gtt gaa tgc cat 768

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255 ttt tgg atg acc ggt gct tgc tgt gaa ccg aga tat tct ctg tcc agg 816

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Arg Tyr Ser Leu Ser Arg 260 265 270 gtg ata gct act aag atg act gca cta ata acc gtc cta gat gat atg 864

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met 275 280 285 atg gac acc tat agc acc act gag gaa gct atg tta ttg gct gaa gct 912

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300 att tat aga tgg gaa gaa aat gcc gct gag tta cta cca ggg tac atg 960

Ile Tyr Arg Trp Glu Glu Asn Ala Ala Glu Leu Leu Pro Gly Tyr Met 305 310 315 320 aaa cat ttt tac tta tac ctg ctt aag aca atc gat tct tgt ggc ggg 1008

Lys His Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Gly 325 330 335 gag ttg ggt ccg aat agg tct ttt agg act ttc tac ttg aaa gag atg 1056

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350 ttg aag gtt ttt gta aga ggt agt tca caa gaa att aaa tgg aga aac 1104

Leu Lys Val Phe Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365 gaa aat tac gtg cca aaa aca att tcc gaa cat tta gaa cac tct gga 1152

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380 ccc aca gtc ggc gcg ttc caa gtc gca tgt tcg tcc ttt gta ggt atg 1200

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400 gga gat aat att act aag gaa tcc ttc gaa tgg ttg ctt acg tac cca 1248

Gly Asp Asn Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415 gaa tta gtt aaa tct ctt atg aat ata gct aga ttg tta aat gat aca 1296

Glu Leu Val Lys Ser Leu Met Asn Ile Ala Arg Leu Leu Asn Asp Thr 420 425 430 gct tct aca aag agg gaa caa acg gct gga cat cat gtt agc acg gtc 1344

Pagina 112

SEQLTXT

Ala Ser Thr Lys Arg Glu Gln Thr Ala Gly His His Val Ser Thr Val 435 440 445 caa tgc tat atg tta aag cat ggc act aca atg gat gag gcc tgt gaa 1392

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Glu 450 455 460 aaa att aaa gaa ctg act gag gat tca tgg aaa gat atg atg gaa tta 14409

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480 tac ctt acc ccc aca gag cat cct aaa ttg gtt gcg caa act att gtt 1488

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Val Ala Gln Thr Ile Val 485 490 495 gat ttt gca aga acg gca gac tat atg tac aaa gag act gat ggg ttt 1536

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510 acc ttc tct cac act ata aaa gac atg ata gca aaa tta ttt gtt gac 1584

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525 cca att tca cta ttc taa 1602

Pro Ile Ser Leu Phe 530 <210> 42 <211> 533 <212> PRT <213> Zea perennis <400> 42

Met Asp Ala Thr Ala Phe His Pro Ser Leu Trp Gly Asp Phe Phe Val 1 5 10 15

Lys Tyr Glu Pro Pro Thr Ala Pro Lys Arg Gly His Met Thr Gln Arg

Ala Glu Leu Leu Lys Glu Glu Val Arg Lys Thr Leu Lys Ala Ala Ala

Asn Gln Ile Lys Asn Ala Leu Asp Leu Ile Ile Thr Leu Gln Arg Leu 60

Gly Leu Asp His His Tyr Glu Asn Glu Ile Ser Glu Leu Leu Arg Phe

Pagina 113

SEQLTXT

65 70 75 80

Val Tyr Ser Ser Ser Asp Tyr Asp Asp Lys Asp Leu Tyr Val Val Ser 85 90 95

Leu Arg Phe Tyr Leu Leu Arg Lys His Gly His Arg Val Ser Ser Asp 100 105 110

Val Phe Met Ser Phe Lys Asp Glu Glu Gly Asn Phe Val Val Asp Asp 115 120 125

Thr Lys Cys Leu Leu Ser Leu Tyr Asn Ala Ala Tyr Leu Met Thr His 130 135 140

Gly Glu Lys Val Leu Asp Glu Ala Ile Thr Phe Thr Arg Arg Gln Leu 145 150 155 160

Glu Ala Leu Leu Leu Asp Pro Leu Glu Pro Ala Leu Ala Asp Glu Val 165 170 175

Tyr Leu Thr Leu Gln Thr Pro Leu Phe Arg Arg Leu Arg Ile Leu Glu 180 185 190

Ala Val Asn Tyr Ile Pro Ile Tyr Gly Lys Glu Ala Gly Arg Asn Glu 195 200 205

Ala Ile Leu Glu Leu Ala Lys Leu Asn Phe Asn Leu Ala Gln Leu Ile 210 215 220

Tyr Cys Glu Glu Leu Lys Glu Val Thr Leu Trp Trp Lys Gln Leu Asn 225 230 235 240

Val Glu Thr Asn Leu Ser Phe Ile Arg Asp Arg Ile Val Glu Cys His 245 250 255

Phe Trp Met Thr Gly Ala Cys Cys Glu Pro Arg Tyr Ser Leu Ser Arg 260 265 270

Val Ile Ala Thr Lys Met Thr Ala Leu Ile Thr Val Leu Asp Asp Met

Pagina 114

SEQLTXT

275 280 285

Met Asp Thr Tyr Ser Thr Thr Glu Glu Ala Met Leu Leu Ala Glu Ala 290 295 300

Ile Tyr Arg Trp Glu Glu Asn Ala Ala Glu Leu Leu Pro Gly Tyr Met 305 310 315 320

Lys His Phe Tyr Leu Tyr Leu Leu Lys Thr Ile Asp Ser Cys Gly Gly 325 330 335

Glu Leu Gly Pro Asn Arg Ser Phe Arg Thr Phe Tyr Leu Lys Glu Met 340 345 350

Leu Lys Val Phe Val Arg Gly Ser Ser Gln Glu Ile Lys Trp Arg Asn 355 360 365

Glu Asn Tyr Val Pro Lys Thr Ile Ser Glu His Leu Glu His Ser Gly 370 375 380

Pro Thr Val Gly Ala Phe Gln Val Ala Cys Ser Ser Phe Val Gly Met 385 390 395 400

Gly Asp Asn Ile Thr Lys Glu Ser Phe Glu Trp Leu Leu Thr Tyr Pro 405 410 415

Glu Leu Val Lys Ser Leu Met Asn Ile Ala Arg Leu Leu Asn Asp Thr 420 425 430

Ala Ser Thr Lys Arg Glu Gln Thr Ala Gly His His Val Ser Thr Val 435 440 445

Gln Cys Tyr Met Leu Lys His Gly Thr Thr Met Asp Glu Ala Cys Glu 450 455 460

Lys Ile Lys Glu Leu Thr Glu Asp Ser Trp Lys Asp Met Met Glu Leu 465 470 475 480

Tyr Leu Thr Pro Thr Glu His Pro Lys Leu Val Ala Gln Thr Ile Val

Pagina 115

SEQLTXT

485 490 495

Asp Phe Ala Arg Thr Ala Asp Tyr Met Tyr Lys Glu Thr Asp Gly Phe 500 505 510

Thr Phe Ser His Thr Ile Lys Asp Met Ile Ala Lys Leu Phe Val Asp 515 520 525

Pro Ile Ser Leu Phe 530

Pagina 116

Claims (43)

ConclusiesConclusions 1. Een eukaryote gastheercel omvattende één of meer heterologe nucleïnezuren, waarbij de heterologe nucleïnezuren het activiteitsniveau in de gastheercel veranderen van a) één of meer enzymen van de mevalonaatroute; en/of b) een prenyltransferase; en waarbij de gastheercel verder een heteroloog [-farneseensynthase omvat en in staat is een sesquiterpeen te maken uit de werking van het B-farneseensynthase.A eukaryotic host cell comprising one or more heterologous nucleic acids, wherein the heterologous nucleic acids alter the activity level in the host cell of a) one or more enzymes of the mevalonate pathway; and/or b) a prenyltransferase; and wherein the host cell further comprises a heterologous β-farnesene synthase and is capable of making a sesquiterpene from the action of the β-farnesene synthase. 2. Gastheercel volgens conclusie 1, waarbij de een of meer enzymen van de mevalonaatroute een 3-hydroxy-3-methylglutaryl-co-enzym-A-reductase (HMGR) is.The host cell of claim 1, wherein the one or more enzymes of the mevalonate pathway is a 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR). 3. Gastheercel volgens conclusie 2, waarbij de HMGR een verkorte HMGR is.The host cell of claim 2, wherein the HMGR is a truncated HMGR. 4. Gastheercel volgens een van de conclusies 1-3, waarbij de prenyltransferase een farnesylpyrofosfaatsynthase (FPPS) is.The host cell of any one of claims 1-3, wherein the prenyl transferase is a farnesyl pyrophosphate synthase (FPPS). 5. Gastheercel volgens conclusie 4, waarbij de FPPS wordt gecodeerd door ERG20.The host cell of claim 4, wherein the FPPS is encoded by ERG20. 6. Gastheercel volgens conclusie 4 of 5, waarbij het heterologe nucleinezuur ten minste één extra kopie omvat van een polynucleotide dat codeert voor de FPPS.The host cell of claim 4 or 5, wherein the heterologous nucleic acid comprises at least one extra copy of a polynucleotide encoding the FPPS. 7. Gastheercel volgens een van de voorgaande conclusies, waarbij de gastheercel ten minste 10% meer (E)-P-farneseen produceert in vergelijking met een controle.A host cell according to any one of the preceding claims, wherein the host cell produces at least 10% more (E)-P-farnesene compared to a control. 8. Gastheercel volgens conclusie 7, waarbij de controle de hoeveelheid (E)-B-farmeseen is die wordt geproduceerd door een gastheercel die Artemisia Annua farneseensynthase tot expressie brengt.The host cell of claim 7, wherein the control is the amount of (E)-B-farmesene produced by a host cell expressing Artemisia Annua farnesene synthase. 9. Gastheercel volgens een van de voorgaande conclusies, waarbij het heterologe f3- farneseensynthase een sequentie omvat met ten minste 70% sequentie-identiteit met SEQ ID NO: 20, 26, 28, 34, 36, 38 of 42 en (2E, 6E)-farnesyldifosfaat kan omzetten tot (E)-beta-farneseen + difosfaat.A host cell according to any one of the preceding claims, wherein the heterologous β-farnesene synthase comprises a sequence with at least 70% sequence identity of SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and (2E, 6E )-farnesyl diphosphate can convert to (E)-beta-farnesene + diphosphate. 10. Gastheercel volgens een van de voorgaande conclusies 1-8, waarbij het heterologe B- farneseensynthase wordt gecodeerd door een polynucleotide dat ten minste 70% sequentie- identiteit heeft met een polynucleotideseguentie van SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35,The host cell of any one of claims 1 to 8, wherein the heterologous β-farnesene synthase is encoded by a polynucleotide having at least 70% sequence identity with a polynucleotide sequence of SEQ ID NO: 19, 21, 23, 25, 27 , 29, 31, 33, 35, 37, 39 of 41 en dat codeert voor een polypeptide dat (2E, 6E)-farnesyldifosfaat omzet in (E)-beta- farneseen + difosfaat.37, 39 or 41 encoding a polypeptide that converts (2E, 6E)-farnesyl diphosphate to (E)-beta-farnesene + diphosphate. 11. Gastheercel volgens een van de voorgaande conclusies 1-9, waarbij het heterologe f3- farneseensynthase een sequentie omvat die is gekozen uit SEQ ID NO: 20, 26, 28, 34, 36, 38 en 42.The host cell of any of claims 1-9, wherein the heterologous β-farnesene synthase comprises a sequence selected from SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42. 12. Gastheercel volgens een van de voorgaande conclusies 1-8, 10, waarbij het heterologe fi- farneseensynthase wordt gecodeerd door een codon-geoptimaliseerde polynucleotidesequentie met ten minste 70% sequentie-identiteit met een polynucleotideseguentie van SEQ ID NO: 19, 21, 23, 25,27, 29,31, 33, 35,37, 39 of 41.The host cell of any of claims 1-8, 10, wherein the heterologous fi-farnesene synthase is encoded by a codon optimized polynucleotide sequence having at least 70% sequence identity with a polynucleotide sequence of SEQ ID NO: 19, 21, 23 , 25.27, 29.31, 33, 35.37, 39 or 41. 13. Een eukaryote gastheercel die één of meer heterologe nucleïnezuren omvat, waarbij de heterologe nucleïnezuren coderen voor: a) een enzym dat 3-hydroxy-3-methylglutaryl-co-enzym-A (HMG-CoA) in mevalonaat omzet; en/of b) een prenyltransferase; en waarbij de gastheercel verder een heteroloog B-farneseensynthase omvat en in staat is een sesquiterpeen te maken uit de werking van het B-farneseensynthase.A eukaryotic host cell comprising one or more heterologous nucleic acids, wherein the heterologous nucleic acids encode: a) an enzyme that converts 3-hydroxy-3-methylglutaryl-coenzyme-A (HMG-CoA) to mevalonate; and/or b) a prenyltransferase; and wherein the host cell further comprises a heterologous β-farnesene synthase and is capable of making a sesquiterpene from the action of the β-farnesene synthase. 14. Gastheercel volgens conclusie 13, waarbij het enzym dat HMG-CoA in mevalonaat omzet, een 3-hydroxy-3-methylglutaryl-co-enzym-A-reductase (HMGR) is.The host cell of claim 13, wherein the enzyme that converts HMG-CoA to mevalonate is a 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR). 15. Gastheercel volgens conclusie 13 of 14, waarbij het enzym dat HMG-CoA in mevalonaat omzet, een verkorte HMGR is.The host cell of claim 13 or 14, wherein the enzyme that converts HMG-CoA to mevalonate is a truncated HMGR. 16. Gastheercel volgens een van de conclusies 13-15, waarbij expressie van het HMGR onder induceerbare controle staat.The host cell of any one of claims 13-15, wherein expression of the HMGR is under inducible control. 17. Gastheercel volgens een van de conclusies 13-15, waarbij de HMGR onder constitutieve controle staat.The host cell of any one of claims 13-15, wherein the HMGR is under constitutive control. 18. Gastheercel volgens een van de conclusies 13-17, waarbij de prenyltransferase een farnesylpyrofosfaatsynthase (FPPS) is.The host cell of any of claims 13-17, wherein the prenyl transferase is a farnesyl pyrophosphate synthase (FPPS). 19. Gastheercel volgens een van de conclusies 13-18, waarbij de prenyltransferase wordt gecodeerd door het ERG20-gen.The host cell of any of claims 13-18, wherein the prenyltransferase is encoded by the ERG20 gene. 20. Gastheercel volgens een van de conclusies 13-19, waarbij de expressie van het prenyltransferase onder induceerbare controle staat.The host cell of any of claims 13-19, wherein the expression of the prenyltransferase is under inducible control. 21. Gastheercel volgens een van de conclusies 13-19, waarbij het prenyltransferase onder constitutieve controle staat.The host cell of any one of claims 13-19, wherein the prenyltransferase is under constitutive control. 22. Gastheercel volgens een van de conclusies 13-21, waarbij het heterologe [-farneseensynthase een sequentie omvat die ten minste 70% sequentie-identiteit heeft met SEQ ID NO: 20, 26, 28, 34, 36, 38 of 42 en (QE, 6E)-farnesyldifosfaat kan omzetten in (E)-beta-farneseen + difosfaat.The host cell of any one of claims 13-21, wherein the heterologous [-farnesene synthase] comprises a sequence having at least 70% sequence identity to SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and ( QE, 6E)-farnesyl diphosphate can convert to (E)-beta-farnesene + diphosphate. 23. Gastheercel volgens een van de voorgaande conclusies 13-21, waarbij het heterologe B- farneseensynthase wordt gecodeerd door een polynucleotide dat ten minste 70% sequentie- identiteit heeft met een polynacleotidesequentie van SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39 of 41 en dat codeert voor een polypeptide dat (2E, 6E)-farnesyldifosfaat omzet in (E)-beta- farneseen + difosfaat.The host cell of any one of claims 13 to 21, wherein the heterologous β-farnesene synthase is encoded by a polynucleotide having at least 70% sequence identity with a polynacleotide sequence of SEQ ID NO: 19, 21, 23, 25, 27 , 29, 31, 33, 35, 37, 39 or 41 encoding a polypeptide that converts (2E, 6E)-farnesyl diphosphate to (E)-beta-farnesene + diphosphate. 24. Gastheercel volgens een van de voorgaande conclusies 13-22, waarbij het heterologe B- farneseensynthase een sequentie omvat gekozen uit SEQ ID NO: 20, 26, 28, 34, 36, 38 en 42.The host cell of any one of claims 13 to 22, wherein the heterologous β-farnesene synthase comprises a sequence selected from SEQ ID NO: 20, 26, 28, 34, 36, 38 and 42. 25. Gastheercel volgens een van de voorgaande conclusies 13-21, 23, waarbij het heterologe p- farneseensynthase wordt gecodeerd door een codon-geoptimaliseerde polynucleotidesequentie met ten minste 70% sequentie-identiteit met een polynucleotidesequentie van SEQ ID NO: 19, 21, 23, 25,27, 29, 31, 33, 35, 37, 39 of 41.The host cell of any one of claims 13-21, 23, wherein the heterologous β-farnesene synthase is encoded by a codon optimized polynucleotide sequence having at least 70% sequence identity with a polynucleotide sequence of SEQ ID NO: 19, 21, 23 , 25,27, 29, 31, 33, 35, 37, 39 or 41. 26. Eukaryote gastheercel volgens een van de voorgaande conclusies 1 tot en met 25, waarbij de eukaryote gastheercel een Saccharomyces cerevisiae cel is.A eukaryotic host cell according to any one of claims 1 to 25, wherein the eukaryotic host cell is a Saccharomyces cerevisiae cell. 27. Een Saccharomyces cerevisiae cel die één of meer heteroloog nucleinezuur omvat, waarbij de heterologe nucleïnezuren coderen voor: a) een verkort 3-hydroxy-3-methylglutaryl-co-enzym-A- reductase (HMGR); en/of b) een prenyltransferase; en een heteroloog p-farneseensynthase en in staat is om een sesquiterpeen te maken uit de werking van het p-farneseensynthase.27. A Saccharomyces cerevisiae cell comprising one or more heterologous nucleic acids, wherein the heterologous nucleic acids encode: a) a truncated 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR); and/or b) a prenyltransferase; and a heterologous β-farnesene synthase and is capable of making a sesquiterpene from the action of the β-farnesene synthase. 28. Een polypeptide dat een sequentie omvat die ten minste 70% sequentie-identiteit heeft met SEQ ID NO: 20, 26, 28, 34, 36, 38 of 42 en (2E, 6E)-farnesyldifosfaat kan omzetten in (E)-bèta farneseen + difosfaat, eventueel is het polypeptide een geïsoleerd polypeptide.28. A polypeptide comprising a sequence having at least 70% sequence identity to SEQ ID NO: 20, 26, 28, 34, 36, 38 or 42 and capable of converting (2E, 6E)-farnesyl diphosphate to (E)- beta farnesene + diphosphate, optionally the polypeptide is an isolated polypeptide. 29. Een codon-geoptimaliseerd polynucleotide dat codeert voor het polypeptide volgens conclusieA codon-optimized polynucleotide encoding the polypeptide of claim 28.28. 30. Een p-farneseensynthase-gen, waarbij het gen ten minste 70% sequentie-identiteit omvat, bij voorkeur ten minste 85% sequentie-identiteit, met meer voorkeur ten minste 90% sequentie- identiteit, met nog meer voorkeur ten minste 95% sequentie-identiteit, met de meeste voorkeur 99 % sequentie-identiteit met een polynucleotidesequentie van SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39 of 41 en waarbij het gen codeert voor een polypeptide voor het omzetten van (2E, 6E) -farnesyldifosfaat tot (E)-beta farneseen + difosfaat.30. A p-farnesene synthase gene, wherein the gene comprises at least 70% sequence identity, preferably at least 85% sequence identity, more preferably at least 90% sequence identity, even more preferably at least 95% sequence identity, most preferably 99% sequence identity to a polynucleotide sequence of SEQ ID NO: 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39 or 41 and wherein the gene encodes a polypeptide for converting (2E, 6E)-farnesyl diphosphate to (E)-beta farnesene + diphosphate. 31. Een vector die het polynucleotide of gen volgens conclusie 29 of 30 omvat.A vector comprising the polynucleotide or gene of claim 29 or 30. 32. Gastheercel die het polypeptide volgens conclusie 28, het polynucleotide volgens conclusie 29 of het [-farneseensynthase-gen van conclusie 30 omvat, waarbij het polypeptide, polynucleotide of gen heteroloog is voor de gastheercel.A host cell comprising the polypeptide of claim 28, the polynucleotide of claim 29 or the β-farnesene synthase gene of claim 30, wherein the polypeptide, polynucleotide or gene is heterologous to the host cell. 33. Gastheercel, eventueel volgens conclusie 32, omvattende de vector volgens conclusie 31.A host cell, optionally according to claim 32, comprising the vector according to claim 31. 34. Gastheercel volgens conclusie 32 of 33, verder omvattende een 3-hydroxy-3-methylglutaryl- co-enzym-A-reductase (HMGR), eventueel een verkort HMGR.The host cell of claim 32 or 33, further comprising a 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR), optionally a truncated HMGR. 35. Gastheercel volgens een van de conclusies 32-34, verder omvattende ten minste twee kopieën van de ERG20-coderende sequentie.The host cell of any one of claims 32-34, further comprising at least two copies of the ERG20 coding sequence. 36. Gastheercel volgens een van de conclusies 32-35, waarbij de gastheercel eukaryoot is.The host cell of any one of claims 32-35, wherein the host cell is eukaryotic. 37. Gastheercel volgens een van de conclusies 32-36, waarbij de gastheercel een gistcel is, bij voorkeur een Saccharomyces cerevisiae cel.A host cell according to any one of claims 32-36, wherein the host cell is a yeast cell, preferably a Saccharomyces cerevisiae cell. 38. Gebruik van één of meer heterologe nucleïnezuren aanwezig in een eukaryote gastheercel of Saccharomyces cerevisiae cel volgens één van de conclusies 1-27, 32-37 voor het veranderen van het activiteitsniveau in genoemde eukaryote gastheercel of Saccharomyces cerevisiae cel, waarbij de heterologe nucleïnezuren het activiteitsniveau veranderen van a) een of meer enzymen van de mevalonaatroute; en/of b) een prenyltransferase; en waarbij de gastheercel verder een heteroloog [3- fameseensynthase omvat en in staat is een sesquiterpeen te maken uit de werking van het p- farneseensynthase.Use of one or more heterologous nucleic acids present in a eukaryotic host cell or Saccharomyces cerevisiae cell according to any one of claims 1-27, 32-37 for altering the level of activity in said eukaryotic host cell or Saccharomyces cerevisiae cell, wherein the heterologous nucleic acids changing activity level of a) one or more enzymes of the mevalonate pathway; and/or b) a prenyltransferase; and wherein the host cell further comprises a heterologous β-farnesene synthase and is capable of making a sesquiterpene from the action of the β-farnesene synthase. 39. Gebruik van een eukaryote gastheercel volgens een van de conclusies 1-27, 32-37 voor het maken van sesquiterpeen en/of farneseen.Use of a eukaryotic host cell according to any one of claims 1-27, 32-37 for making sesquiterpene and/or farnesene. 40. Gebruik van de Saccharomyces cerevisiae cel volgens conclusie 27 voor het maken van sesquiterpeen en/of farneseen.Use of the Saccharomyces cerevisiae cell according to claim 27 for making sesquiterpene and/or farnesene. 41. Werkwijze voor het produceren van B-farneseen, waarbij de werkwijze omvat het kweken van een eukaryote gastheercel of Saccharomyces cerevisiae cel volgens een van de conclusies 1-27, 32- 37, onder omstandigheden en met een geschikt substraat, waarbij het substraat wordt gemetaboliseerd tot B-farneseen.A method of producing β-farnesene, the method comprising culturing a eukaryotic host cell or Saccharomyces cerevisiae cell according to any one of claims 1-27, 32-37, under conditions and with a suitable substrate, the substrate being metabolized to β-farnesene. 42. Werkwijze volgens conclusie 41, waarbij de eukaryote gastheercel of Saccharomyces cerevisiae cel verder een of meer van de enzymen omvat die zijn gekozen uit Acetoacetyl-coA- thiolase (acetyl-coA-acetyltransferase); Hydroxymethyglutaryl-CoA-synthase (HMGS); Mevalonaatkinase (ERG12); Fosfovalonaatkinase (ERGR); Difosfomevalonaat decarboxylase (ERG19); Isopentenylpyrofosfaatisomerase (IDI), of een combinatie daarvan.The method of claim 41, wherein the eukaryotic host cell or Saccharomyces cerevisiae cell further comprises one or more of the enzymes selected from Acetoacetyl-coA thiolase (acetyl-coA acetyltransferase); Hydroxymethylglutaryl-CoA synthase (HMGS); Mevalonate kinase (ERG12); Phosphovalonate Kinase (ERGR); Diphosphomevalonate decarboxylase (ERG19); Isopentenyl pyrophosphate isomerase (IDI), or a combination thereof. 43. Werkwijze volgens conclusie 41 of 42, waarbij het substraat glucose is.The method of claim 41 or 42, wherein the substrate is glucose.