LOCUS       sequence-247137-        3887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid complementing ADH3 from Komagataella phaffii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3887)
  AUTHORS   Liu Y, Bai C, Liu Q, Xu Q, Qian Z, Peng Q, Yu J, Xu M, Zhou X, Zhang
            Y, Cai M
  TITLE     Engineered ethanol-driven biosynthetic system for improving 
            production of acetyl-CoA derived drugs in Crabtree-negative yeast.
  JOURNAL   Metab Eng. 2019 Jul;54:275-284. doi: 10.1016/j.ymben.2019.05.001. 
            Epub 2019 May 9.
  PUBMED    31077813
REFERENCE   2  (bases 1 to 3887)
  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."; date: "2019-07"; volume: "54:275-284. doi"; pages: " 
            10.1016/j.ymben.2019.05.001. Epub 2019 May 9"
COMMENT     Sequence Label: pZ_P-GAP-ADH3
FEATURES             Location/Qualifiers
     source          1..3887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..512
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             1593..1622
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1638..1655
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1735..1981
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1996..2409
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2416..2463
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2482..2856
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      2922..3169
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2938..2956)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3244..3832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3324..3343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttaagggat tttggtcatg catgagatca gatctttttt gtagaaatgt cttggtgtcc
       61 tcgtccaatc aggtagccat ctctgaaata tctggctccg ttgcaactcc gaacgacctg
      121 ctggcaacgt aaaattctcc ggggtaaaac ttaaatgtgg agtaatggaa ccagaaacgt
      181 ctcttccctt ctctctcctt ccaccgcccg ttaccgtccc taggaaattt tactctgctg
      241 gagagcttct tctacggccc ccttgcagca atgctcttcc cagcattacg ttgcgggtaa
      301 aacggaggtc gtgtacccga cctagcagcc cagggatgga aaagtcccgg ccgtcgctgg
      361 caataatagc gggcggacgc atgtcatgag attattggaa accaccagaa tcgaatataa
      421 aaggcgaaca cctttcccaa ttttggtttc tcctgaccca aagactttaa atttaattta
      481 tttgtcccta tttcaatcaa ttgaacaact atatgtctcc aactatccca actacacaaa
      541 aggctgttat cttcgagacc aacggcggtc ccctagagta caaggacatt ccagtcccaa
      601 agccaaagtc aaacgaactt ttgatcaacg ttaagtactc cggtgtctgt cacactgatt
      661 tgcacgcctg gaagggtgac tggccattgg acaacaagct tcctttggtt ggtggtcacg
      721 aaggtgctgg tgtcgttgtc gcttacggtg agaacgtcac tggatgggag atcggtgact
      781 acgctggtat caaatggttg aacggttctt gtttgaactg tgagtactgt atccaaggtg
      841 ctgaatccag ttgtgccaag gctgacctgt ctggtttcac ccacgacgga tctttccagc
      901 agtatgctac tgctgatgcc acccaagccg ccagaattcc aaaggaggct gacttggctg
      961 aagttgcccc aattctgtgt gctggtatca ccgtttacaa ggctcttaag accgctgact
     1021 tgcgtattgg ccaatgggtt gccatttctg gtgctggtgg aggactgggt tctcttgccg
     1081 ttcaatacgc caaggctctg ggtttgagag ttttgggtat tgatggtggt gccgacaagg
     1141 gtgaatttgt caagtccttg ggtgctgagg tcttcgtcga cttcactaag actaaggacg
     1201 tcgttgctga agtccaaaag ctcaccaacg gtggtccaca cggtgttatt aacgtctccg
     1261 tttccccaca tgctatcaac caatctgtcc aatacgttag aactttgggt aaggttgttt
     1321 tggttggtct gccatctggt gccgttgtca actctgacgt tttctggcac gttctgaagt
     1381 ccatcgagat caagggatct tacgttggaa acagagagga cagtgccgag gccatcgact
     1441 tgttcaccag aggtttggtc aaggctccta tcaagattat cggtctgtct gaacttgcta
     1501 aggtctacga acagatggag gctggtgcca tcatcggtag atacgttgtg gacacttcca
     1561 aaagtcgagc cgcggcggcc gccagctttc tagaacaaaa actcatctca gaagaggatc
     1621 tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttgtagcctt agacatgact
     1681 gttcctcagt tcaagttggg cacttacgag aagaccggtc ttgctagatt ctaatcaaga
     1741 ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt
     1801 gtaacctata tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc
     1861 ctgatcagcc tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc
     1921 gagtttgatg tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag
     1981 accttcgttt gtgcggatcc cccacacacc atagcttcaa aatgtttcta ctcctttttt
     2041 actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac
     2101 agcatactaa atttcccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg
     2161 tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa
     2221 atttttatca cgtttctttt tcttgaaaat tttttttttt gatttttttc tctttcgatg
     2281 acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt
     2341 tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc
     2401 taatctaagg gcggtgttga caattaatca tcggcatagt atatcggcat agtataatac
     2461 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     2521 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     2581 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     2641 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     2701 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     2761 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     2821 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tccgacgcgg cccgacgggt
     2881 ccgaggcctc ggagatccgt cccccttttc ctttgtcgat atcatgtaat tagttatgtc
     2941 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     3001 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     3061 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     3121 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca agctggagac
     3181 caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcac
//
