LOCUS       sequence-405765-        7122 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 
            ADH2. URA3 selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7122)
  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 7122)
  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: pHYX138
FEATURES             Location/Qualifiers
     source          1..7122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             42..62
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(402..421)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             699..710
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             753..809
                     /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             816..836
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             855..911
                     /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             951..971
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1727..2530)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(2531..2746)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2770..2789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2889..2911
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2949..2967)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3008..4348
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    3338..3385
                     /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)."
     promoter        4375..4479
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4762..4781)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4773..5588
                     /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     complement(4846..4865)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5836..6424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6325..6344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6578..6595
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6607..6623
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6607..6623
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgtaatacac aatggataaa gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg
       61 tcgaattggg taccgccggc ggctccggcc ccgggatggt cagcaagggc gaggaactct
      121 tcaccggcgt cgtccctatc ctcgtcgagc ttgacggcga cgtcaacggc cacaagttct
      181 ccgtcagcgg cgagggcgag ggcgacgcca cctacggcaa gctcaccctc aagctcatct
      241 gcaccaccgg caagctccct gtcccttggc ctaccctcgt caccaccctc ggctacggcc
      301 tccagtgctt cgcccgctac cctgaccaca tgaagcagca cgacttcttc aagtccgcca
      361 tgcctgaggg ctacgtccaa gagcgcacca ttttcttcaa ggacgacggc aactacaaga
      421 cccgcgctga ggtcaagttc gagggagaca ccctcgtcaa ccgcatcgag ctgaagggca
      481 tcgacttcaa ggaagatggc aacatcctcg gccacaagct cgagtacaac tacaactccc
      541 acaacgtcta catcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagattc
      601 gccacaacat cgagagagga ggcggcggat ctggcggagg acccggggcc ctgaagaagg
      661 agctgcaggc caacaagaag gagctggcgc agctgaagtg ggagctgcag gccctgaaga
      721 aggagctcgc tcaatgttta aacggctctg gcgccaccaa cttctctctc ctcaaacagg
      781 ctggcgacgt tgaagaaaac cctggccctg atatcgaaaa tctttatttt caaggtgcga
      841 tgggaggctc tggcgccacc aacttctctc tcctcaaaca ggctggcgac gttgaagaaa
      901 accctggccc tcatttaaat atggacaagg ccgagctgat ccgcgagccc cccaagaaga
      961 agcgcaaggt ggagctgggc accgccggcg gctccggcga gcagctggag aagaagctgc
     1021 aggccctgga gaagaagctg gcccagctgg agtggaagaa ccaggccctg gagaagaagc
     1081 tggcccaacc cgggggcgga ggaggctcag gcggaggcgg ctctgacaag cagaagaacg
     1141 gcatcaaggc caacttcaag attcgccaca acatcgagga cggcggcgtc cagctcgccg
     1201 accactacca gcagaacacc cctatcggcg acggccctgt cctcctccct gacaaccact
     1261 acctcagcta ccagtccgcc ctctccaagg accctaacga gaagcgcgac cacatggtcc
     1321 tgctcgagtt cgtcaccgcc gctggcatca ccctcggcat ggacgagctg tacaagtaga
     1381 tagattaatt taaacagtat atctacagtt ttatatatat atatatatat atatacatat
     1441 ataaagaaac ctgtgcgttt tttgtatttt caaatacatt tagttttggc gccacttcta
     1501 taaaaggtct catgatattg ttacaatatc aatttttctt cgtttctcat gccctgtttc
     1561 tatgatagag gatgaattta ttattaacgg cactaacaac gtacatatag cgcccgggcc
     1621 gcatctgtgc ggtatttcac accgcatagg gtaataactg atataattaa attgaagctc
     1681 taatttgtga gtttagtata catgcattta cttataatac agttttttag ttttgctggc
     1741 cgcatcttct caaatatgct tcccagcctg cttttctgta acgttcaccc tctaccttag
     1801 catcccttcc ctttgcaaat agtcctcttc caacaataat aatgtcagat cctgtagaga
     1861 ccacatcatc cacggttcta tactgttgac ccaatgcgtc tcccttgtca tctaaaccca
     1921 caccgggtgt cataatcaac caatcgtaac cttcatctct tccacccatg tctctttgag
     1981 caataaagcc gataacaaaa tctttgtcgc tcttcgcaat gtcaacagta cccttagtat
     2041 attctccagt agatagggag cccttgcatg acaattctgc taacatcaaa aggcctctag
     2101 gttcctttgt tacttcttct gccgcctgct tcaaaccgct aacaatacct gggcccacca
     2161 caccgtgtgc attcgtaatg tctgcccatt ctgctattct gtatacaccc gcagagtact
     2221 gcaatttgac tgtattacca atgtcagcaa attttctgtc ttcgaagagt aaaaaattgt
     2281 acttggcgga taatgccttt agcggcttaa ctgtgccctc catcgaaaaa tcagtcaaaa
     2341 tatccacatg tgtttttagt aaacaaattt tgggacctaa tgcttcaact aactccagta
     2401 attccttggt ggtacgaaca tccaatgaag cacacaagtt tgtttgcttt tcgtgcatga
     2461 tattaaatag cttggcagca acaggactag gatgagtagc agcacgttcc ttatatgtag
     2521 ctttcgacat gatttatctt cgtttcctgc aggtttttgt tctgtgcagt tgggttaaga
     2581 atactgggca atttcatgtt tcttcaacac tacatatgcg tatatatacc aatctaagtc
     2641 tgtgctcctt ccttcgttct tccttctgtt cggagattac cgaatcaaaa aaatttcaag
     2701 gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga attgaaaagg tggtatggtg
     2761 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     2821 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     2881 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     2941 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3001 ttagtatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga
     3061 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc
     3121 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg
     3181 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat
     3241 acaggcaaca cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg
     3301 ttgcattttc ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct
     3361 attctctaga aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac
     3421 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata
     3481 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc
     3541 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca
     3601 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc
     3661 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga
     3721 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact
     3781 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta
     3841 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag
     3901 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc
     3961 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta
     4021 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa
     4081 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa
     4141 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca
     4201 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg
     4261 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     4321 ctacaaaatg aagcacagat gcttcgttca ggtggcactt ttcggggaaa tgtgcgcgga
     4381 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4441 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtcg ggaagatgcg
     4621 tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccac gttgtgtctc
     4681 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     4741 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt
     4801 gctcgaggcc gcatttaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     4861 gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc
     4921 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     4981 tccgtttaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     5041 ctcctgatga tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat
     5101 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     5161 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     5221 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     5281 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac
     5341 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     5401 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     5461 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     5521 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     5581 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     5641 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5701 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggt
     5761 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5821 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5881 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5941 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6001 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6061 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6121 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6181 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6241 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6301 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6361 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6421 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6481 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6541 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6601 gtcacacagg aaacagctat gaccatggcc cgggctatct aaaaattgcc ttatgatccg
     6661 tctctccggt tacagcctgt gtaactgatt aatcctgcct ttctaatcac cattctaatg
     6721 ttttaattaa gggattttgt cttcattaac ggctttcgct cataaaaatg ttatgacgtt
     6781 ttgcccgcag gcgggaaacc atccacttca cgagactgat ctcctctgcc ggaacaccgg
     6841 gcatctccaa cttataagtt ggagaaataa gagaatttca gattgagaga atgaaaaaaa
     6901 aaaaaaaaaa aggcagagga gagcatagaa atggggttca ctttttggta aagctatagc
     6961 atgcctatca catataaata gagtgccagt agcgactttt ttcacactcg aaatactctt
     7021 actactgctc tcttgttgtt tttatcactt cttgtttctt cttggtaaat agaatatcaa
     7081 gctacaaaaa gcatacaatc aactatcaac tattaactat at
//
