LOCUS       sequence-248171-        3560 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 3560)
  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 3560)
  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-eGFP
FEATURES             Location/Qualifiers
     source          1..3560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..539
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             546..1265
                     /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             1293..1322
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1338..1355
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1435..1681
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1696..2109
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2116..2163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2182..2556
                     /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      2622..2869
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2638..2656)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2944..3532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3024..3043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat gagatcagat
       61 cttttttgta gaaatgtctt ggtgtcctcg tccaatcagg tagccatctc tgaaatatct
      121 ggctccgttg caactccgaa cgacctgctg gcaacgtaaa attctccggg gtaaaactta
      181 aatgtggagt aatggaacca gaaacgtctc ttcccttctc tctccttcca ccgcccgtta
      241 ccgtccctag gaaattttac tctgctggag agcttcttct acggccccct tgcagcaatg
      301 ctcttcccag cattacgttg cgggtaaaac ggaggtcgtg tacccgacct agcagcccag
      361 ggatggaaaa gtcccggccg tcgctggcaa taatagcggg cggacgcatg tcatgagatt
      421 attggaaacc accagaatcg aatataaaag gcgaacacct ttcccaattt tggtttctcc
      481 tgacccaaag actttaaatt taatttattt gtccctattt caatcaattg aacaactata
      541 ccaccatggg ttctaaaggt gaagaattat tcactggtgt tgtcccaatt ttggttgaat
      601 tagatggtga tgttaatggt cacaaatttt ctgtctccgg tgaaggtgaa ggtgatgcta
      661 cttacggtaa attgacctta aaatttattt gtactactgg taaattgcca gttccatggc
      721 caaccttagt cactactttc ggttatggtg ttcaatgttt tgcgagatac ccagatcata
      781 tgaaacaaca tgactttttc aagtctgcca tgccagaagg ttatgttcaa gaaagaacta
      841 tttttttcaa agatgacggt aactacaaga ccagagctga agtcaagttt gaaggtgata
      901 ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa agaagatggt aacattttag
      961 gtcacaaatt ggaatacaac tataactctc acaatgttta catcatggct gacaaacaaa
     1021 agaatggtat caaagttaac ttcaaaatta gacacaacat tgaagatggt tctgttcaat
     1081 tagctgacca ttatcaacaa aatactccaa ttggtgatgg tccagtcttg ttaccagaca
     1141 accattactt atccactcaa tctgccttat ccaaagatcc aaacgaaaag agagaccaca
     1201 tggtcttgtt agaatttgtt actgctgctg gtattaccca tggtatggat gaattgtaca
     1261 aatagcgagc cgcggcggcc gccagctttc tagaacaaaa actcatctca gaagaggatc
     1321 tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttgtagcctt agacatgact
     1381 gttcctcagt tcaagttggg cacttacgag aagaccggtc ttgctagatt ctaatcaaga
     1441 ggatgtcaga atgccatttg cctgagagat gcaggcttca tttttgatac ttttttattt
     1501 gtaacctata tagtatagga ttttttttgt cattttgttt cttctcgtac gagcttgctc
     1561 ctgatcagcc tatctcgcag ctgatgaata tcttgtggta ggggtttggg aaaatcattc
     1621 gagtttgatg tttttcttgg tatttcccac tcctcttcag agtacagaag attaagtgag
     1681 accttcgttt gtgcggatcc cccacacacc atagcttcaa aatgtttcta ctcctttttt
     1741 actcttccag attttctcgg actccgcgca tcgccgtacc acttcaaaac acccaagcac
     1801 agcatactaa atttcccctc tttcttcctc tagggtgtcg ttaattaccc gtactaaagg
     1861 tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt cgtcgaaaaa ggcaataaaa
     1921 atttttatca cgtttctttt tcttgaaaat tttttttttt gatttttttc tctttcgatg
     1981 acctcccatt gatatttaag ttaataaacg gtcttcaatt tctcaagttt cagtttcatt
     2041 tttcttgttc tattacaact ttttttactt cttgctcatt agaaagaaag catagcaatc
     2101 taatctaagg gcggtgttga caattaatca tcggcatagt atatcggcat agtataatac
     2161 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     2221 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     2281 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     2341 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     2401 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     2461 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     2521 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tccgacgcgg cccgacgggt
     2581 ccgaggcctc ggagatccgt cccccttttc ctttgtcgat atcatgtaat tagttatgtc
     2641 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     2701 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     2761 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     2821 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca agctggagac
     2881 caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3541 tttgatcttt tctacggggt
//
