LOCUS       sequence-248266-        3327 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 3327)
  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 3327)
  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_lacO-cP-AOX1-eGFP
FEATURES             Location/Qualifiers
     source          1..3327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    77..93
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             294..1013
                     /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             1075..1104
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1120..1137
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1217..1463
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1478..1891
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1898..1945
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1964..2338
                     /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      2404..2651
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2420..2438)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2726..3314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2806..2825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
       61 tgagatctgt gtggaattgt gagcggataa caatttcaca cactaacccc tacttgacag
      121 caatatataa acagaaggaa gctgccctgt cttaaacctt tttttttatc atcattatta
      181 gcttactttc ataattgcga ctggttccaa ttgacaagct tttgatttta acgactttta
      241 acgacaactt gagaagatca aaaaacaact aattattcga aggatcctac accatgggtt
      301 ctaaaggtga agaattattc actggtgttg tcccaatttt ggttgaatta gatggtgatg
      361 ttaatggtca caaattttct gtctccggtg aaggtgaagg tgatgctact tacggtaaat
      421 tgaccttaaa atttatttgt actactggta aattgccagt tccatggcca accttagtca
      481 ctactttcgg ttatggtgtt caatgttttg cgagataccc agatcatatg aaacaacatg
      541 actttttcaa gtctgccatg ccagaaggtt atgttcaaga aagaactatt tttttcaaag
      601 atgacggtaa ctacaagacc agagctgaag tcaagtttga aggtgatacc ttagttaata
      661 gaatcgaatt aaaaggtatt gattttaaag aagatggtaa cattttaggt cacaaattgg
      721 aatacaacta taactctcac aatgtttaca tcatggctga caaacaaaag aatggtatca
      781 aagttaactt caaaattaga cacaacattg aagatggttc tgttcaatta gctgaccatt
      841 atcaacaaaa tactccaatt ggtgatggtc cagtcttgtt accagacaac cattacttat
      901 ccactcaatc tgccttatcc aaagatccaa acgaaaagag agaccacatg gtcttgttag
      961 aatttgttac tgctgctggt attacccatg gtatggatga attgtacaaa tagacgtggc
     1021 ccagccggcc gtctcggatc ggtacctcga gccgcggcgg ccgccagctt tctagaacaa
     1081 aaactcatct cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga
     1141 gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg
     1201 tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt
     1261 catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt
     1321 ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg
     1381 taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc actcctcttc
     1441 agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca ccatagcttc
     1501 aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg catcgccgta
     1561 ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc tctagggtgt
     1621 cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc
     1681 ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa attttttttt
     1741 ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa cggtcttcaa
     1801 tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac ttcttgctca
     1861 ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat catcggcata
     1921 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     1981 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     2041 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     2101 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     2161 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     2221 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     2281 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     2341 cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt tcctttgtcg
     2401 atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     2461 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     2521 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     2581 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     2641 ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2701 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2761 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2821 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2881 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2941 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3001 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3061 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3121 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3181 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3241 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3301 aaaaaggatc tcaagaagat cctttga
//
