LOCUS       sequence-242585-        4058 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 4058)
  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 4058)
  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-ACS1-eGFP
FEATURES             Location/Qualifiers
     source          1..4058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1143..1862
                     /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             1863..1880
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1960..2206
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2221..2632
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2640..2687
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2706..3080
                     /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      3146..3393
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3162..3180)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3468..4056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3548..3567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
       61 gattttggtc atgagatcag atctaaaacc accagctagt acagaggtaa aaaagtagat
      121 aagcaatcct accagcaaaa tcatctggct cagcggttat gttggttagg gtctaagttt
      181 tgaccaattt gcttgtatgt gcgttaaaaa taggacagtt gggggacaat agcgcgactg
      241 gatttggtag acagggcttg gttgaaggtt gacaattgac caaccaagca ataccggtag
      301 tgggcaaatg aaaaaagacc accatccgcc atccgacagc acactaccgt catcccctct
      361 tccgtaagac caaaacatca ttctctcttc ccctcctagg ttggtgattc gcgacccatt
      421 cctcaactcc agagttgttc attttacaat ggggaagtgg gatatgattt cgttcctcaa
      481 ttgacttatt tgtcatgctt tttcggagct tgaagcatat cggtctctgt accctgagcc
      541 tttcgcatta gtcggagtgc ggatatgtta tgggtcaggg tctgtatctt gaattattgt
      601 aaaattgaaa gaaaggatct aaggattgtt tttccctttg aatgtgtatc ttaagaaatt
      661 gagtttcacc agcgtgttgt ttactaccag cgtctctctt ctgtgtgatg gccatactat
      721 actacatacc actttactcc atttccgata ataagtttca cttcatcgtc tttacctcct
      781 tgactccgtg cggctccaac tctaaatcta ttagatgatc atgaggggca gcatttctcg
      841 taatatccca cagagatacg tcaaatccca ttgttccggg tatatgccct cagtaaagac
      901 atggcgtcca gcatcttgta ctcacatctc gtttgagcgg gtgacgggag gggtatatgc
      961 atctattttt cttgagtctt gactctgaag taggtctcgt acaatatgtg acggagagca
     1021 ttcgtgttat aaaaaggtaa gaatggtccc ccattttctt ctttctttct ttctttctac
     1081 tatctttata aagttttttt ttacaaagga tacgacttag ttgttgatca attgaattca
     1141 ccatgggttc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
     1201 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
     1261 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
     1321 ccttagtcac tactttcggt tatggtgttc aatgttttgc gagataccca gatcatatga
     1381 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
     1441 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
     1501 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
     1561 acaaattgga atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga
     1621 atggtatcaa agttaacttc aaaattagac acaacattga agatggttct gttcaattag
     1681 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
     1741 attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg
     1801 tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaat
     1861 agcatcatca tcatcatcat tgagtttgta gccttagaca tgactgttcc tcagttcaag
     1921 ttgggcactt acgagaagac cggtcttgct agattctaat caagaggatg tcagaatgcc
     1981 atttgcctga gagatgcagg cttcattttt gatacttttt tatttgtaac ctatatagta
     2041 taggattttt tttgtcattt tgtttcttct cgtacgagct tgctcctgat cagcctatct
     2101 cgcagctgat gaatatcttg tggtaggggt ttgggaaaat cattcgagtt tgatgttttt
     2161 cttggtattt cccactcctc ttcagagtac agaagattaa gtgagacctt cgtttgtgcg
     2221 gatcccccac acaccatagc ttcaaaatgt ttctactcct tttttactct tccagatttt
     2281 ctcggactcc gcgcatcgcc gtaccacttc aaaacaccca agcacagcat actaaatttt
     2341 ccctctttct tcctctaggg tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa
     2401 agagaccgcc tcgtttcttt ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt
     2461 ctttttcttg aaattttttt ttttagtttt tttctctttc agtgacctcc attgatattt
     2521 aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca
     2581 acttttttta cttcttgttc attagaaaga aagcatagca atctaatcta aggggcggtg
     2641 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     2701 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     2761 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     2821 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     2881 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     2941 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     3001 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     3061 tggccgagga gcaggactga cacgtccgac ggcggcccac gggtcccagg cctcggagat
     3121 ccgtccccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     3181 cctcccccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     3241 cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     3301 cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     3361 aaggttttgg gacgctcgaa ggctttaatt tgcaagctgg agaccaacat gtgagcaaaa
     3421 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3481 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3541 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3601 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3661 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3721 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3781 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3841 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3901 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3961 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4021 aagcagcaga ttacgcgcag aaaaaaagga tctcaaga
//
