LOCUS       sequence-257296-        4009 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 4009)
  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 4009)
  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-ICL1-eGFP
FEATURES             Location/Qualifiers
     source          1..4009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1046..1765
                     /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             1766..1783
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1863..2109
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2124..2537
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2544..2591
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2610..2984
                     /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      3050..3297
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3066..3084)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3372..3960)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3452..3471)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actcacgtta agggattttg gtcatgagat cagatcttca tctaacactt tgtatagcac
       61 atcgtaccag tttatcaaaa tccaacaagt tttcctctgg gtactgaaat tcgattagta
      121 ataaacgggc gcacttggtg atctcctcat cctgttcact agccttcttt tcgcttaact
      181 tcatcaacga gatcaactcg agtctgcgag cagagaactg tttccaaaga gtttcatcgc
      241 tgaaattcag cttctttcga ataaaatgtg tcactcccac tttattactc gtggtgttat
      301 ttgttttcca cgaactagtt gaatcgccat ctttactacc gtcctgggtg acatccggcg
      361 aattcgggac aaatgtgctg ttccggtagc ttgtaggaag cggcatccgt agggcaatat
      421 acgactatag cttctaaagc gtagtacaat gaaatgttcg aaggaacaac aaacggattt
      481 gtttttcgta ggctcaaccc gttgaggtgt aactctttag cgaaagggta agattgattg
      541 ttcgaagtag ggcctcaaag ggaaagagaa aaaaaaaaat acaccaagag ttacgtaagc
      601 atatattttt tacgtaaagc atgattgaat ttcagcagta ttgtttaaca aggctgatgt
      661 cgtgtgccaa tcaaaacaaa agagattcgc ataatgccat aattggggtg tgtgggcgcc
      721 ccctaaaacg tctttctcat catcatctgc aacccccatc gaacctcatt aaatcacatg
      781 acttgtgcga tcctcggtca actcgttccg tgcacccatt ccaccccggg ctgaccaacg
      841 caaggttctc cgagagtccg ctaccccaga tttatatcag caaccagtca cctttttccg
      901 ggcacgactc tatatgccct ggaaaaccgg agacgatgag cctgactgta aaaggtgaca
      961 gaacccccaa ctctggttaa tctcttcaac aaatacttta ttttctttca attcaaagaa
     1021 cacagtatca agtatatcaa gaaccatggg ttctaaaggt gaagaattat tcactggtgt
     1081 tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt ctgtctccgg
     1141 tgaaggtgaa ggtgatgcta cttacggtaa attgacctta aaatttattt gtactactgg
     1201 taaattgcca gttccatggc caaccttagt cactactttc ggttatggtg ttcaatgttt
     1261 tgcgagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca tgccagaagg
     1321 ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga ccagagctga
     1381 agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta ttgattttaa
     1441 agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc acaatgttta
     1501 catcatggct gacaaacaaa agaatggtat caaagttaac ttcaaaatta gacacaacat
     1561 tgaagatggt tctgttcaat tagctgacca ttatcaacaa aatactccaa ttggtgatgg
     1621 tccagtcttg ttaccagaca accattactt atccactcaa tctgccttat ccaaagatcc
     1681 aaacgaaaag agagaccaca tggtcttgtt agaatttgtt actgctgctg gtattaccca
     1741 tggtatggat gaattgtaca aatagcatca tcatcatcat cattgagttt gtagccttag
     1801 acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt gctagattct
     1861 aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt tttgatactt
     1921 ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct tctcgtacga
     1981 gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg ggtttgggaa
     2041 aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag tacagaagat
     2101 taagtgagac cttcgtttgt gcggatcccc cacacaccat agcttcaaaa tgtttctact
     2161 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
     2221 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
     2281 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
     2341 caataaaaat ttttatcacg tttctttttc ttgaaaattt ttttttttga tttttttctc
     2401 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
     2461 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
     2521 tagcaatcta atctaagggc ggtgttgaca attaatcatc ggcatagtat atcggcatag
     2581 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     2641 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     2701 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     2761 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     2821 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     2881 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     2941 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtc cgacgcggcc
     3001 cgacgggtcc gaggcctcgg agatccgtcc cccttttcct ttgtcgatat catgtaatta
     3061 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
     3121 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     3181 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     3241 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     3301 ctggagacca acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3361 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3421 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3481 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3541 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3601 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3661 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3721 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3781 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3841 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3901 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3961 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaa
//
