LOCUS       sequence-247141-        4841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid complementing ACS1 from  Komagataella phaffii.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4841)
  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 4841)
  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-ACS1
FEATURES             Location/Qualifiers
     source          1..4841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..512
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             2547..2576
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2592..2609
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2689..2935
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2950..3363
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3370..3417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3436..3810
                     /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      3876..4123
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3892..3910)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4198..4786)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4278..4297)
                     /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 atatgccatt agataacgaa cacttacttc
      541 atgaaaattc cattgaccca ccaaagggat tctttgaaag acaccctgga actcctaata
      601 taccaggcgg ttgggaagaa tacttgaagc tgtacaatca gtccatcgag aacccctcaa
      661 agttttttgg agaaaaagca aaggaattct tgtcatgggc tactcctttc actgacgctc
      721 gttacccacc tggtaatgga tttcagaatg gtgactccgc cgcttggttt ctgaatggtg
      781 agttgaacgc gtcgtacaac tgtgttgata gacatgcttt aaagaatcca gacaaacctg
      841 ccattattta tgaggctgat gaacctaatc aaggccgtac ggttacctat ggagagttgc
      901 tgaaggatgt ttgtcgaatt gcccaagtat tgactgacct gggtgtgaaa aagggtgaca
      961 ctgttgctgt ttacctgcct atggttccag aagctatcac cactttattg gctatcgtta
     1021 gaatcggtgc tatccactct gttgtcttcg caggtttttc agctggttct ctacgtgatc
     1081 gtatattgga tgctgattct agaattgtta tcacttctga tgaatctctg agaggtggga
     1141 agatcatcga gactaagaag attgttgacg aggctctgaa gtcttgccca gatgttcgta
     1201 atgtgctggt cttcaaaaga acaggtacac cacatcttcc atgggttgag ggtcgtgatc
     1261 tttggtggca cgaggaaatc attaagcatg ttccgtactc tcccccagtg aatgttagat
     1321 ctgaagatac ttcatttttg ctttacactt ctggctctac cggaaagcct aaaggtatcc
     1381 agcattcaac tgctggctac ttactgggag ctcttttgac caccaagtat gtctttgatg
     1441 ttcagggtga tgatatttta ttcactgctg gtgatgtggg ctggatcaca gggcattctt
     1501 atgtagttta cggtccactt ttaaacgggg ctacgacagt tgtttttgag ggcaccccag
     1561 cttacccaga ctattcacgt tattgggata tcgttgacaa acacaaagtt actcagtttt
     1621 atgtagcacc aactgctctt aggttgctga agagagctgg tagcaagtat gtccagaatc
     1681 atgatttgtc ttcaatcagg gttttgggtt ccgttggtga acctatagcc gctgaagttt
     1741 gggaatggta caacgagtat gttggaagag gaaaagctca tatttgtgat acgtattggc
     1801 aaacagagac tggttctcac attattgctc caatagctgg tgtgtcaaag accaaaccag
     1861 gttcagcatc tttccccttc ttcggtattg atccggttat tctagatgct actactggag
     1921 aggaactcaa aggtaataat gttgaaggtg ttttggctat cagaaatcca tggccatcta
     1981 tggctagaac agtctggaag gactacaacc gtttcctgga tacatatctc aggccatatg
     2041 aaggttatta cttcactggt gatggagctg ccagagatca ggaaggattt tattgggttc
     2101 tgggtagagt tgatgatgtt gttaatgtgt caggtcacag attgtctact gccgagattg
     2161 aaagcgctct aatcgaacac aatttggtag gagagtctgc tgtcgtcgga ttccctgacg
     2221 agctgactgg ttctgctgtg gccgcgtttg tgtctttgaa gaaggacgtc gacaatccag
     2281 cggaagtgaa aaaggagtta atccttactg tcagaaaaga gattggacca ttcgctgcac
     2341 ctaaactcat catcttggta agtgatcttc caaagaccag atcaggtaag ataatgagac
     2401 gtattctcag aaaggttttg gctggagagg aagactctct gggcgacatt tcaactcttt
     2461 caaacccttc gattgtggaa gagataatct ctaccgttaa aagggatgcc cgcaaacctc
     2521 gagccgcggc ggccgccagc tttctagaac aaaaactcat ctcagaagag gatctgaata
     2581 gcgccgtcga ccatcatcat catcatcatt gagtttgtag ccttagacat gactgttcct
     2641 cagttcaagt tgggcactta cgagaagacc ggtcttgcta gattctaatc aagaggatgt
     2701 cagaatgcca tttgcctgag agatgcaggc ttcatttttg atactttttt atttgtaacc
     2761 tatatagtat aggatttttt ttgtcatttt gtttcttctc gtacgagctt gctcctgatc
     2821 agcctatctc gcagctgatg aatatcttgt ggtaggggtt tgggaaaatc attcgagttt
     2881 gatgtttttc ttggtatttc ccactcctct tcagagtaca gaagattaag tgagaccttc
     2941 gtttgtgcgg atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt
     3001 ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata
     3061 ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga
     3121 aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt
     3181 atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc gatgacctcc
     3241 cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt
     3301 gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct
     3361 aagggcggtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3421 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     3481 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     3541 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     3601 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     3661 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     3721 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     3781 gtgcacttcg tggccgagga gcaggactga cacgtccgac gcggcccgac gggtccgagg
     3841 cctcggagat ccgtccccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     3901 acattcacgc cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     3961 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt
     4021 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac
     4081 cttgcttgag aaggttttgg gacgctcgaa ggctttaatt tgcaagctgg agaccaacat
     4141 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4201 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4261 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4321 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4381 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4441 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4501 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4561 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4621 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4681 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4741 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4801 cttttctacg gggtctgacg ctcagtggaa cgaaaactca c
//
