LOCUS       sequence-247145-        3999 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 3999)
  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 3999)
  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-ALD-eGFP
FEATURES             Location/Qualifiers
     source          1..3999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1068..1787
                     /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             1788..1805
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1885..2131
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2146..2559
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2566..2613
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2632..3006
                     /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      3072..3319
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3088..3106)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3394..3982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3474..3493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
       61 atcagaccag cagtttaact acgcaaatcc acaggaattt ctacatcaca ataccaatgg
      121 taataccacg acgtcaagga atggaaacga cgacttggag gaagacttcg tcaacctctt
      181 gcggagtacc cgaggctaag acaataagaa gaaaaaaaaa agaaaagcgg tgggggaggg
      241 attattaaat aaggattatg taaccccagg gtaccgttct atacatattt aaggattatt
      301 taggacaatc gatgaaatcg gcatcaaact ggatgggagt atagtgtccg gataatcgga
      361 taaatcatct tgcgaggagc cgcttggttg gttggtgaga ggagtgaaat atgtgtctcc
      421 tcacccaaga atcgcgatat cagcaccctg tgggggacac tattggcctc cctcccaaac
      481 cttcgatgtg gtagtgcttt attatattga ttacattgat tacatagcta aaccctgcct
      541 ggttgcaagt tgagctccga attccaatat tagtaaaatg cctgcaagat aacctcggta
      601 tggcgtccga ccccgcttaa ttattttaac tcctttccaa cgaggacttc gtaatttttg
      661 attagggagt tgagaaacgg ggggtcttga tacctcctcg atttcagatc ccaccccctc
      721 tcagtcccaa gtgggacccc cctcggccgt gaaatgcgcg cactttagtt tttttcgcat
      781 gtaaacgccg gtgtccgtca attaaaagtc gcagactagg gtgaacttta ccatttttgt
      841 cgcactccgt ctcctcggaa taggggtgta gtaattctgc agtagtgcaa tttttacccc
      901 gccaaggggg ggcgaaaaga gacgacctca tcacgcattc tccagtcgct ctctacgcct
      961 acagcaccga cgtagttaac tttctcccat atataaagca attgccattc ccctgaaaac
     1021 tttaacctct gctttttctt gatttttcct tgcccaaaga aaagaccatg ggttctaaag
     1081 gtgaagaatt attcactggt gttgtcccaa ttttggttga attagatggt gatgttaatg
     1141 gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc tacttacggt aaattgacct
     1201 taaaatttat ttgtactact ggtaaattgc cagttccatg gccaacctta gtcactactt
     1261 tcggttatgg tgttcaatgt tttgcgagat acccagatca tatgaaacaa catgactttt
     1321 tcaagtctgc catgccagaa ggttatgttc aagaaagaac tatttttttc aaagatgacg
     1381 gtaactacaa gaccagagct gaagtcaagt ttgaaggtga taccttagtt aatagaatcg
     1441 aattaaaagg tattgatttt aaagaagatg gtaacatttt aggtcacaaa ttggaataca
     1501 actataactc tcacaatgtt tacatcatgg ctgacaaaca aaagaatggt atcaaagtta
     1561 acttcaaaat tagacacaac attgaagatg gttctgttca attagctgac cattatcaac
     1621 aaaatactcc aattggtgat ggtccagtct tgttaccaga caaccattac ttatccactc
     1681 aatctgcctt atccaaagat ccaaacgaaa agagagacca catggtcttg ttagaatttg
     1741 ttactgctgc tggtattacc catggtatgg atgaattgta caaatagcat catcatcatc
     1801 atcattgagt ttgtagcctt agacatgact gttcctcagt tcaagttggg cacttacgag
     1861 aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat
     1921 gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt
     1981 cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata
     2041 tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac
     2101 tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatcc cccacacacc
     2161 atagcttcaa aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca
     2221 tcgccgtacc acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc
     2281 tagggtgtcg ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt
     2341 tctttttctt cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat
     2401 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg
     2461 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt
     2521 cttgctcatt agaaagaaag catagcaatc taatctaagg gcggtgttga caattaatca
     2581 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag
     2641 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
     2701 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
     2761 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
     2821 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
     2881 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
     2941 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
     3001 gactgacacg tccgacgcgg cccgacgggt ccgaggcctc ggagatccgt cccccttttc
     3061 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     3121 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     3181 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     3241 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     3301 ctcgaaggct ttaatttgca agctggagac caacatgtga gcaaaaggcc agcaaaaggc
     3361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3961 gcgcagaaaa aaaggatctc aagaagatcc tttgatctt
//
