LOCUS       sequence-248137-        4059 bp    DNA     circular SYN 15-APR-2025
DEFINITION  yeast plasmid overexpressing eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4059)
  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 4059)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..4059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      17..263
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        278..691
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        698..745
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             764..1138
                     /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      1204..1451
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1220..1238)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(1526..2114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1606..1625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3259..3978
                     /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             3979..3996
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt
       61 catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt
      121 ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg
      181 taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc
      241 agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc
      301 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
      361 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
      421 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc
      481 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
      541 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa
      601 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
      661 ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat catcggcata
      721 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
      781 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
      841 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
      901 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
      961 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     1021 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     1081 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     1141 cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt tcctttgtcg
     1201 atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     1261 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     1321 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     1381 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     1441 ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2161 aaaactcacg ttaagggatt ttggtcatga gatcagatct aaaaccacca gctagtacag
     2221 aggtaaaaaa gtagataagc aatcctacca gcaaaatcat ctggctcagc ggttatgttg
     2281 gttagggtct aagttttgac caatttgctt gtatgtgcgt taaaaatagg acagttgggg
     2341 gacaatagcg cgactggatt tggtagacag ggcttggttg aaggttgaca attgaccaac
     2401 caagcaatac cggtagtggg caaatgaaaa aagaccacca tccgccatcc gacagcacac
     2461 taccgtcatc ccctcttccg taagaccaaa acatcattct ctcttcccct cctaggttgg
     2521 tgattcgcga cccattcctc aactccagag ttgttcattt tacaatgggg aagtgggata
     2581 tgatttcgtt cctcaattga cttatttgtc atgctttttc ggagcttgaa gcatatcggt
     2641 ctctgtaccc tgagcctttc gcattagtcg gagtgcggat atgttatggg tcagggtctg
     2701 tatcttgaat tattgtaaaa ttgaaagaaa ggatctaagg attgtttttc cctttgaatg
     2761 tgtatcttaa gaaattgagt ttcaccagcg tgttgtttac taccagcgtc tctcttctgt
     2821 gtgatggcca tactatacta cataccactt tactccattt ccgataataa gtttcacttc
     2881 atcgtcttta cctccttgac tccgtgcggc tccaactcta aatctattag atgatcatga
     2941 ggggcagcat ttctcgtaat atcccacaga gatacgtcaa atcccattgt tccgggtata
     3001 tgccctcagt aaagacatgg cgtccagcat cttgtactca catctcgttt gagcgggtga
     3061 cgggaggggt atatgcatct atttttcttg agtcttgact ctgaagtagg tctcgtacaa
     3121 tatgtgacgg agagcattcg tgttataaaa aggtaagaat ggtcccccat tttcttcttt
     3181 ctttctttct ttctactatc tttataaagt ttttttttac aaaggatacg acttagttgt
     3241 tgatcaattg aattcaccat gggttctaaa ggtgaagaat tattcactgg tgttgtccca
     3301 attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt
     3361 gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg
     3421 ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg ttttgcgaga
     3481 tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt
     3541 caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag
     3601 tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat
     3661 ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg
     3721 gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat
     3781 ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc
     3841 ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa
     3901 aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg
     3961 gatgaattgt acaaatagca tcatcatcat catcattgag tttgtagcct tagacatgac
     4021 tgttcctcag ttcaagttgg gcacttacga gaagaccgg
//
