LOCUS       sequence-405740-        7779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Saccharomyces cerevisiae vector for the expression of polycistronic 
            genes. Expression controled by the auto-inducible yeast promoter 
            PCK1. LEU2 selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7779)
  AUTHORS   Geistodt-Kiener A, Totozafy JC, Le Goff G, Vergne J, Sakai K, 
            Ouazzani J, Mouille G, Viaud M, O'Connell RJ, Dallery JF
  TITLE     Yeast-based heterologous production of the colletochlorin family of 
            fungal secondary metabolites.
  JOURNAL   Metab Eng. 2023 Oct 18:S1096-7176(23)00142-8. doi: 
            10.1016/j.ymben.2023.10.002.
  PUBMED    37863177
REFERENCE   2  (bases 1 to 7779)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab Eng. 
            2023 Oct 18:S1096-7176(23)00142-8. doi: 
            10.1016/j.ymben.2023.10.002."
COMMENT     Sequence Label: pHYX137
FEATURES             Location/Qualifiers
     source          1..7779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(247..267)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(811..827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(811..827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(839..856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1010..1598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1090..1109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1846..2661)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPHLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2569..2588
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2653..2672
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        complement(2955..3059)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3086..4428
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(4049..4096)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     4469..4487
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4525..4547)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4665..5070
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5071..6165
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     6422..6439
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6423..6439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(7117..7137)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7177..7233)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(7252..7272)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(7279..7335)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(7378..7389)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     7667..7686
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gcactggagg ccgtagccga gggtggtgac gagggtaggc caagggacag ggagcttgcc
       61 ggtggtgcag atgagcttga gggtgagctt gccgtaggtg gcgtcgccct cgccctcgcc
      121 gctgacggag aacttgtggc cgttgacgtc gccgtcaagc tcgacgagga tagggacgac
      181 gccggtgaag agttcctcgc ccttgctgac catcccgggg ccggagccgc cggcggtacc
      241 caattcgacc tttctcttct tttttggagg ctcgggaatt aattccgctt tatccatgtt
      301 gttattttat tatggaataa ttagttgcgt gagtttggtt tttttttttt tatttttgag
      361 ttaaacaaat ataattattg tttaagctga aataaagata tatatgtata tctgggaaag
      421 gctgttggtt atctgtctta cttctgtata tttaagaagt aagtagaaat actgattaca
      481 catggaagag aagttacaca cattacaaag acttctctgc ccttatatac cttttttttg
      541 tggtcttcta tactctgccc aatgttgcta tccccagcca agccttactt aatacacaaa
      601 cgtaccattg tccaaccaga agcgtctaag ctttacaatc ctgtttgggc cctttatccg
      661 cctatccata atgttaagca gcctaaaact tgctagtcta gccggcatct ttcccttgta
      721 taacttaaaa tacccggagc ttcagtgata tgtagaacac agccgcgccg gatgaaagga
      781 agctcggttt tttcctatgc ccgggccatg gtcatagctg tttcctgtgt gacgcttcct
      841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1441 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgacctttt
     1681 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     1741 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcgtcaagt
     1801 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     1861 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     1921 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     1981 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2041 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2101 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2161 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2221 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     2281 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     2341 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     2401 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttaaac ggaccatctc
     2461 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     2521 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     2581 tttataccca tataaatcag catccatgtt ggaatttaaa tgcggcctcg agcaagacgt
     2641 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     2701 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     2761 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     2821 tcccgaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     3001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     3061 acatttcccc gaaaagtgcc acctgaacga agcatctgtg cttcattttg tagaacaaaa
     3121 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     3181 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     3241 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3301 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     3361 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     3421 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     3481 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     3541 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     3601 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     3661 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     3721 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     3781 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     3841 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     3901 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     3961 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca
     4021 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact
     4081 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
     4141 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
     4201 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct
     4261 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg
     4321 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat
     4381 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat actaagaaac
     4441 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4501 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4561 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4621 cgggtgtcgg ggctggctta actatgcggc aggggatcct ctagaactgt gggaatactc
     4681 aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat
     4741 aacgagaaca cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg
     4801 ttaatttcag aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag
     4861 gaaaggtgag agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa
     4921 tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa
     4981 taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat
     5041 atatatatat ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg
     5101 ccaggtgacc acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt
     5161 tctgatgttc gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct
     5221 atcgatgcta caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat
     5281 gccgttttgt taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa
     5341 caaggtttac taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac
     5401 tttgcatccg actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact
     5461 gacttcgttg ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac
     5521 gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc
     5581 acaagaatgg ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg
     5641 gataaagcta atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc
     5701 aagaacgaat tccctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc
     5761 ctagttaaga acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat
     5821 attatctccg atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc
     5881 ttggcctctt tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct
     5941 gctccagatt tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg
     6001 atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa
     6061 aaggttttgg atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa
     6121 gtcggtgatg ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt
     6181 ttatgatatt tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta
     6241 caaaatggaa tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc
     6301 aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc
     6361 aacgtgataa ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcaccg
     6421 ttgtaaaacg acggccagtg gcccgggcct ctagaggcat tttcaacatc gtattttccg
     6481 aagcgttgca taatgaaaat gtgataataa ggcagtgagc tactttgtca caacaaagta
     6541 cgagaacaga tgaggttccg tggtgttttt ggggcaaccg cactatttgg agcgctgtat
     6601 atatttatat cctactacaa aaaatcataa atgacactga attggacaca tgctagcgtc
     6661 aaataccttg cctggtaaag ttgtgtgcta gtgtctcccg tcttctgtct acttgtacag
     6721 ctcgtccatg ccgagggtga tgccagcggc ggtgacgaac tcgagcagga ccatgtggtc
     6781 gcgcttctcg ttagggtcct tggagagggc ggactggtag ctgaggtagt ggttgtcagg
     6841 gaggaggaca gggccgtcgc cgataggggt gttctgctgg tagtggtcgg cgagctggac
     6901 gccgccgtcc tcgatgttgt ggcgaatctt gaagttggcc ttgatgccgt tcttctgctt
     6961 gtcagagccg cctccgcctg agcctcctcc gcccccgggt tgggccagct tcttctccag
     7021 ggcctggttc ttccactcca gctgggccag cttcttctcc agggcctgca gcttcttctc
     7081 cagctgctcg ccggagccgc cggcggtgcc cagctccacc ttgcgcttct tcttgggggg
     7141 ctcgcggatc agctcggcct tgtccatatt taaatgaggg ccagggtttt cttcaacgtc
     7201 gccagcctgt ttgaggagag agaagttggt ggcgccagag cctcccatcg caccttgaaa
     7261 ataaagattt tcgatatcag ggccagggtt ttcttcaacg tcgccagcct gtttgaggag
     7321 agagaagttg gtggcgccag agccgtttaa acattgagcg agctccttct tcagggcctg
     7381 cagctcccac ttcagctgcg ccagctcctt cttgttggcc tgcagctcct tcttcagggc
     7441 cccgggtcct ccgccagatc cgccgcctcc tctctcgatg ttgtggcgaa tcttgaagtt
     7501 ggccttgatg ccgttcttct gcttgtcggc ggtgatgtag acgttgtggg agttgtagtt
     7561 gtactcgagc ttgtggccga ggatgttgcc atcttccttg aagtcgatgc ccttcagctc
     7621 gatgcggttg acgagggtgt ctccctcgaa cttgacctca gcgcgggtct tgtagttgcc
     7681 gtcgtccttg aagaaaatgg tgcgctcttg gacgtagccc tcaggcatgg cggacttgaa
     7741 gaagtcgtgc tgcttcatgt ggtcagggta gcgggcgaa
//
