LOCUS       sequence-248159-        3940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid overexpressing eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3940)
  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 3940)
  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-AOX1-eGFP
FEATURES             Location/Qualifiers
     source          1..3940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..994
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1000..1719
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             1720..1737
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1817..2063
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2078..2491
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2498..2545
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2564..2938
                     /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      3004..3251
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3020..3038)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3326..3914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3406..3425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gatcagatct
       61 aacatccaaa gacgaaaggt tgaatgaaac ctttttgcca tccgacatcc acaggtccat
      121 tctcacacat aagtgccaaa cgcaacagga ggggatacac tagcagcaga ccgttgcaaa
      181 cgcaggacct ccactcctct tctcctcaac acccactttt gccatcgaaa aaccagccca
      241 gttattgggc ttgattggag ctcgctcatt ccaattcctt ctattaggct actaacacca
      301 tgactttatt agcctgtcta tcctggcccc cctggcgagg ttcatgtttg tttatttccg
      361 aatgcaacaa gctccgcatt acacccgaac atcactccag atgagggctt tctgagtgtg
      421 gggtcaaata gtttcatgtt ccccaaatgg cccaaaactg acagtttaaa cgctgtcttg
      481 gaacctaata tgacaaaagc gtgatctcat ccaagatgaa ctaagtttgg ttcgttgaaa
      541 tgctaacggc cagttggtca aaaagaaact tccaaaagtc ggcataccgt ttgtcttgtt
      601 tggtattgat tgacgaatgc tcaaaaataa tctcattaat gcttagcgca gtctctctat
      661 cgcttctgaa ccccggtgca cctgtgccga aacgcaaatg gggaaacacc cgctttttgg
      721 atgattatgc attgtctcca cattgtatgc ttccaagatt ctggtgggaa tactgctgat
      781 agcctaacgt tcatgatcaa aatttaactg ttctaacccc tacttgacag caatatataa
      841 acagaaggaa gctgccctgt cttaaacctt tttttttatc atcattatta gcttactttc
      901 ataattgcga ctggttccaa ttgacaagct tttgatttta acgactttta acgacaactt
      961 gagaagatca aaaaacaact aattattcga aacgagacca tgggttctaa aggtgaagaa
     1021 ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
     1081 ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattt
     1141 atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttcggttat
     1201 ggtgttcaat gttttgcgag atacccagat catatgaaac aacatgactt tttcaagtct
     1261 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
     1321 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
     1381 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac
     1441 tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa
     1501 attagacaca acattgaaga tggttctgtt caattagctg accattatca acaaaatact
     1561 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccac tcaatctgcc
     1621 ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct
     1681 gctggtatta cccatggtat ggatgaattg tacaaatagc atcatcatca tcatcattga
     1741 gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg
     1801 tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt
     1861 catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt
     1921 ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg
     1981 taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc
     2041 agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc
     2101 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     2161 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     2221 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc
     2281 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     2341 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa
     2401 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
     2461 ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat catcggcata
     2521 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     2581 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     2641 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     2701 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     2761 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     2821 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     2881 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     2941 cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt tcctttgtcg
     3001 atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     3061 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     3121 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     3181 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     3241 ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3301 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3361 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3421 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3481 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3541 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3601 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3661 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3721 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3781 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3841 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3901 aaaaaggatc tcaagaagat cctttgatct tttctacggg
//
