LOCUS       sequence-247143-        4210 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 4210)
  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 4210)
  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-ADH3-eGFP
FEATURES             Location/Qualifiers
     source          1..4210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1275..1994
                     /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             1995..2012
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2092..2338
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2353..2766
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2773..2820
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2839..3213
                     /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      3279..3526
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3295..3313)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3601..4189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3681..3700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatcc
       61 gcagcgtttt ctgacggtac tagaggactc ttaggggaag gtagaatcaa taaagatcat
      121 attaggtaag caaattttgg atggaatagg agactaggtg tggatgcgcg atctcgccaa
      181 attgcacgac cagagtggat gccggatggt ggtaaaccgt ttcttccttt ttaccaccca
      241 agtgcgagtg aaacacccca tggctgctct ccgattgccc ctctacaggc ataagggtgt
      301 gactttgtgg gcttgaattt tacaccccct ccaacttttc tcgcatcaat tgatcctgtt
      361 accaatattg catgcccgga ggagacttgc cccctaattt cgcggcgtcg tcccggatcg
      421 cagggtgaga ctgtagagac cccacatagt gacaatgatt atgtaagaag aggggggtga
      481 ttcggccggc tatcgaactc taacaactag gggggtgaac aatgcccagc agtcctcccc
      541 actctttgac aaatcagtat caccgattaa caccccaaat cttattctca acggtccctc
      601 atccttgcac ccctctttgg acaaatggca gttagcattg gtgcactgac tgactgccca
      661 accttaaacc caaatttctt agaaggggcc catctagtta gcgaggggtg aaaaattcct
      721 ccatcggaga tgtattgacc gtaagttgct gcttaaaaaa aatcagttca gatagcgaga
      781 cttttttgat ttcgcaacgg gagtgcctgt tccattcgat tgcaattctc accccttctg
      841 cccagtcctg ccaattgccc atgaatctgc taatttcgtt gattcccacc cccctttcca
      901 actccacaaa ttgtccaatc tcgttttcca tttgggagaa tctgcatgtc gactacataa
      961 agcgaccggt gtccgaaaag atctgtgtag ttttcaacat tttgtgctcc ccccgctgtt
     1021 tgaaaacggg ggtgagcgct ctccggggtg cgaattcgtg cccaattcct ttcaccctgc
     1081 ctattgtaga cgtcaacccg catctggtgc gaatatagcg cacccccaat gatcacacca
     1141 acaattggtc cacccctccc caatctctaa tattcacaat tcacctcact ataaataccc
     1201 ctgtcctgct cccaaattct tttttccttc ttccatcagc tactagcttt tatcttattt
     1261 actttacgaa aaccatgggt tctaaaggtg aagaattatt cactggtgtt gtcccaattt
     1321 tggttgaatt agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag
     1381 gtgatgctac ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag
     1441 ttccatggcc aaccttagtc actactttcg gttatggtgt tcaatgtttt gcgagatacc
     1501 cagatcatat gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag
     1561 aaagaactat ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg
     1621 aaggtgatac cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta
     1681 acattttagg tcacaaattg gaatacaact ataactctca caatgtttac atcatggctg
     1741 acaaacaaaa gaatggtatc aaagttaact tcaaaattag acacaacatt gaagatggtt
     1801 ctgttcaatt agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt
     1861 taccagacaa ccattactta tccactcaat ctgccttatc caaagatcca aacgaaaaga
     1921 gagaccacat ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg
     1981 aattgtacaa atagcatcat catcatcatc attgagtttg tagccttaga catgactgtt
     2041 cctcagttca agttgggcac ttacgagaag accggtcttg ctagattcta atcaagagga
     2101 tgtcagaatg ccatttgcct gagagatgca ggcttcattt ttgatacttt tttatttgta
     2161 acctatatag tataggattt tttttgtcat tttgtttctt ctcgtacgag cttgctcctg
     2221 atcagcctat ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa atcattcgag
     2281 tttgatgttt ttcttggtat ttcccactcc tcttcagagt acagaagatt aagtgagacc
     2341 ttcgtttgtg cggatccccc acacaccata gcttcaaaat gtttctactc cttttttact
     2401 cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc
     2461 atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt
     2521 ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt
     2581 tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc
     2641 tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
     2701 cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa
     2761 tctaagggcg gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     2821 aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc
     2881 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     2941 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     3001 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     3061 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     3121 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3181 tgcgtgcact tcgtggccga ggagcaggac tgacacgtcc gacgcggccc gacgggtccg
     3241 aggcctcgga gatccgtccc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     3301 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     3361 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     3421 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     3481 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcaagc tggagaccaa
     3541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3781 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4021 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4201 gatcttttct
//
