LOCUS       sequence-242589-        4952 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 4952)
  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 4952)
  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-HXT1-eGFP
FEATURES             Location/Qualifiers
     source          1..4952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1959..2678
                     /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             2679..2696
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2776..3022
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3037..3448
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3456..3503
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3522..3896
                     /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      3962..4209
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3978..3996)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4284..4872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4364..4383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg agggtaccca attgattaag
      961 ttcagtgaaa tttcaaaccg ctatacacaa caggacaact ttgagtttag aaaaatccga
     1021 tgtagtgtaa cggctagcac ggtccgcttt caccgggcag acccgggttc gactcccggc
     1081 atcggagttt atttttccat ttcgttcttt agagtattct cctcagcatg cccccctgaa
     1141 tttttccttt tttccatgtg tcccattttt ccactttttt tacagttttc ctcgtgatgt
     1201 taattggcta cacaaaagct gccacacgaa accttaatca cgaaaaacta tacagccttc
     1261 actaatccgt agccccataa tatgttgtcc acgtgctgtt gggtactacc tgtagactct
     1321 cataccccac tccgtctttc tccaacaatt aacgcagtac cgagatttat cagcagactc
     1381 aaattgggca aactctgtat ttttccttgc ccgcataatt tatgggtctc aggcctccac
     1441 gtttcctgtt tacttgaaga atattggctg cggaaaaagt ggtaaggaca accccctttt
     1501 aattggatcc agtttttccg aaatgttccg atccgtacgt catctccgaa gccgtacatt
     1561 ttcactcaat ctacgtagct ttggactcag cgctcctgga attgccagga cagtttactt
     1621 gagttgatat tcccttgtag attgtgtgct tctttttcca aaatttgagg cttcgtttga
     1681 aaagtggaat ctggtcgcta gatcacttca tgcctatttt tcacggaaaa ataagtggta
     1741 ctatgcaccc cttaaaccta aagaaaaacg gaaaaattac cccaaaacct ggtgatgttt
     1801 ttcgcccctt tctttttatc cgagtttttc ttttttcttg tctgccaaat tcctctcctg
     1861 accttagcgt ccccggaaaa aattaactac ttaaggaccg aatgagcccc agcttttccc
     1921 cttctcttca ttattcccca taatataatg aattcaccat gggttctaaa ggtgaagaat
     1981 tattcactgg tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat
     2041 tttctgtctc cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta
     2101 tttgtactac tggtaaattg ccagttccat ggccaacctt agtcactact ttcggttatg
     2161 gtgttcaatg ttttgcgaga tacccagatc atatgaaaca acatgacttt ttcaagtctg
     2221 ccatgccaga aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca
     2281 agaccagagc tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag
     2341 gtattgattt taaagaagat ggtaacattt taggtcacaa attggaatac aactataact
     2401 ctcacaatgt ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa
     2461 ttagacacaa cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc
     2521 caattggtga tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct
     2581 tatccaaaga tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg
     2641 ctggtattac ccatggtatg gatgaattgt acaaatagca tcatcatcat catcattgag
     2701 tttgtagcct tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt
     2761 cttgctagat tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc
     2821 atttttgata cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt
     2881 tcttctcgta cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt
     2941 aggggtttgg gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca
     3001 gagtacagaa gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca
     3061 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
     3121 cacttcaaaa cacccaagca cagcatacta aattttccct ctttcttcct ctagggtgtc
     3181 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
     3241 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaat tttttttttt
     3301 agtttttttc tctttcagtg acctccattg atatttaagt taataaacgg tcttcaattt
     3361 ctcaagtttc agtttcattt ttcttgttct attacaactt tttttacttc ttgttcatta
     3421 gaaagaaagc atagcaatct aatctaaggg gcggtgttga caattaatca tcggcatagt
     3481 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     3541 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     3601 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     3661 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     3721 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     3781 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     3841 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     3901 tccgacggcg gcccacgggt cccaggcctc ggagatccgt cccccttttc ctttgtcgat
     3961 atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     4021 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     4081 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     4141 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     4201 ttaatttgca agctggagac caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4441 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4681 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4921 aactcacgtt aagggatttt ggtcatgaga tc
//
