LOCUS       sequence-248169-        4953 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 4953)
  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 4953)
  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-HXT1-eGFP
FEATURES             Location/Qualifiers
     source          1..4953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..988
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             2007..2726
                     /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             2727..2744
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2824..3070
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3085..3498
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3505..3552
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3571..3945
                     /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      4011..4258
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4027..4045)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4333..4921)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4413..4432)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagatcag atctaacatc
       61 caaagacgaa aggttgaatg aaaccttttt gccatccgac atccacaggt ccattctcac
      121 acataagtgc caaacgcaac aggaggggat acactagcag cagaccgttg caaacgcagg
      181 acctccactc ctcttctcct caacacccac ttttgccatc gaaaaaccag cccagttatt
      241 gggcttgatt ggagctcgct cattccaatt ccttctatta ggctactaac accatgactt
      301 tattagcctg tctatcctgg cccccctggc gaggttcatg tttgtttatt tccgaatgca
      361 acaagctccg cattacaccc gaacatcact ccagatgagg gctttctgag tgtggggtca
      421 aatagtttca tgttccccaa atggcccaaa actgacagtt taaacgctgt cttggaacct
      481 aatatgacaa aagcgtgatc tcatccaaga tgaactaagt ttggttcgtt gaaatgctaa
      541 cggccagttg gtcaaaaaga aacttccaaa agtcggcata ccgtttgtct tgtttggtat
      601 tgattgacga atgctcaaaa ataatctcat taatgcttag cgcagtctct ctatcgcttc
      661 tgaaccccgg tgcacctgtg ccgaaacgca aatggggaaa cacccgcttt ttggatgatt
      721 atgcattgtc tccacattgt atgcttccaa gattctggtg ggaatactgc tgatagccta
      781 acgttcatga tcaaaattta actgttctaa cccctacttg acagcaatat ataaacagaa
      841 ggaagctgcc ctgtcttaaa cctttttttt tatcatcatt attagcttac tttcataatt
      901 gcgactggtt ccaattgaca agcttttgat tttaacgact tttaacgaca acttgagaag
      961 atcaaaaaac aactaattat tcgaaacgag ggtacccaat tgattaagtt cagtgaaatt
     1021 tcaaaccgct atacacaaca ggacaacttt gagtttagaa aaatccgatg tagtgtaacg
     1081 gctagcacgg tccgctttca ccgggcagac ccgggttcga ctcccggcat cggagtttat
     1141 ttttccattt cgttctttag agtattctcc tcagcatgcc cccctgaatt tttccttttt
     1201 tccatgtgtc ccatttttcc acttttttta cagttttcct cgtgatgtta attggctaca
     1261 caaaagctgc cacacgaaac cttaatcacg aaaaactata cagccttcac taatccgtag
     1321 ccccataata tgttgtccac gtgctgttgg gtactacctg tagactctca taccccactc
     1381 cgtctttctc caacaattaa cgcagtaccg agatttatca gcagactcaa attgggcaaa
     1441 ctctgtattt ttccttgccc gcataattta tgggtctcag gcctccacgt ttcctgttta
     1501 cttgaagaat attggctgcg gaaaaagtgg taaggacaac ccccttttaa ttggatccag
     1561 tttttccgaa atgttccgat ccgtacgtca tctccgaagc cgtacatttt cactcaatct
     1621 acgtagcttt ggactcagcg ctcctggaat tgccaggaca gtttacttga gttgatattc
     1681 ccttgtagat tgtgtgcttc tttttccaaa atttgaggct tcgtttgaaa agtggaatct
     1741 ggtcgctaga tcacttcatg cctatttttc acggaaaaat aagtggtact atgcacccct
     1801 taaacctaaa gaaaaacgga aaaattaccc caaaacctgg tgatgttttt cgcccctttc
     1861 tttttatccg agtttttctt ttttcttgtc tgccaaattc ctctcctgac cttagcgtcc
     1921 ccggaaaaaa ttaactactt aaggaccgaa tgagccccag cttttcccct tctcttcatt
     1981 attccccata atataatgaa ttcaccatgg gttctaaagg tgaagaatta ttcactggtg
     2041 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
     2101 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
     2161 gtaaattgcc agttccatgg ccaaccttag tcactacttt cggttatggt gttcaatgtt
     2221 ttgcgagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
     2281 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     2341 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     2401 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     2461 acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca
     2521 ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     2581 gtccagtctt gttaccagac aaccattact tatccactca atctgcctta tccaaagatc
     2641 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     2701 atggtatgga tgaattgtac aaatagcatc atcatcatca tcattgagtt tgtagcctta
     2761 gacatgactg ttcctcagtt caagttgggc acttacgaga agaccggtct tgctagattc
     2821 taatcaagag gatgtcagaa tgccatttgc ctgagagatg caggcttcat ttttgatact
     2881 tttttatttg taacctatat agtataggat tttttttgtc attttgtttc ttctcgtacg
     2941 agcttgctcc tgatcagcct atctcgcagc tgatgaatat cttgtggtag gggtttggga
     3001 aaatcattcg agtttgatgt ttttcttggt atttcccact cctcttcaga gtacagaaga
     3061 ttaagtgaga ccttcgtttg tgcggatccc ccacacacca tagcttcaaa atgtttctac
     3121 tcctttttta ctcttccaga ttttctcgga ctccgcgcat cgccgtacca cttcaaaaca
     3181 cccaagcaca gcatactaaa tttcccctct ttcttcctct agggtgtcgt taattacccg
     3241 tactaaaggt ttggaaaaga aaaaagagac cgcctcgttt ctttttcttc gtcgaaaaag
     3301 gcaataaaaa tttttatcac gtttcttttt cttgaaaatt tttttttttg atttttttct
     3361 ctttcgatga cctcccattg atatttaagt taataaacgg tcttcaattt ctcaagtttc
     3421 agtttcattt ttcttgttct attacaactt tttttacttc ttgctcatta gaaagaaagc
     3481 atagcaatct aatctaaggg cggtgttgac aattaatcat cggcatagta tatcggcata
     3541 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     3601 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     3661 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     3721 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     3781 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     3841 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     3901 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt ccgacgcggc
     3961 ccgacgggtc cgaggcctcg gagatccgtc ccccttttcc tttgtcgata tcatgtaatt
     4021 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
     4081 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
     4141 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
     4201 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcaa
     4261 gctggagacc aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4321 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4381 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4441 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4501 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4561 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4621 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4681 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4741 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4801 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4861 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4921 agaagatcct ttgatctttt ctacggggtc tga
//
