LOCUS       sequence-242592-        6298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LacI-Mit1AD on pGAPZ_ A expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6298)
  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 6298)
  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-GAP-LacI-Mit1AD
FEATURES             Location/Qualifiers
     source          1..6298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..483
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             3978..4007
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4023..4040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4119..4365
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        4380..4791
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        4798..4845
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4864..5238
                     /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      5304..5551
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5320..5338)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(5626..6214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5706..5725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcttttt tgtagaaatg tcttggtgtc ctcgtccaat caggtagcca tctctgaaat
       61 atctggctcc gttgcaactc cgaacgacct gctggcaacg taaaattctc cggggtaaaa
      121 cttaaatgtg gagtaatgga accagaaacg tctcttccct tctctctcct tccaccgccc
      181 gttaccgtcc ctaggaaatt ttactctgct ggagagcttc ttctacggcc cccttgcagc
      241 aatgctcttc ccagcattac gttgcgggta aaacggaggt cgtgtacccg acctagcagc
      301 ccagggatgg aaaagtcccg gccgtcgctg gcaataatag cgggcggacg catgtcatga
      361 gattattgga aaccaccaga atcgaatata aaaggcgaac acctttccca attttggttt
      421 ctcctgaccc aaagacttta aatttaattt atttgtccct atttcaatca attgaacaac
      481 tatggatcca ccatgggtgt taagccagtt actttgtatg acgttgctga atacgctgga
      541 gtttcctacc aaactgtctc tagagttgtt aatcaagctt ctcatgtctc cgctaagact
      601 agagagaagg ttgaggctgc tatggctgaa ttgaactata ttccaaatag agttgctcag
      661 cagttggctg gaaagcaatc tttgttgatt ggagtcgcta cttcttcttt ggctttgcat
      721 gctccatctc agattgttgc tgctattaag tccagagctg accagttggg agcttctgtt
      781 gttgtttcta tggttgagag atctggagtt gaggcttgca aggctgctgt tcataacttg
      841 ttggctcaga gagtttctgg attgattatt aattacccat tggacgatca agacgctatt
      901 gccgttgagg ccgcttgtac caacgtccca gctttgttct tggacgtttc cgatcaaact
      961 ccaattaatt ctattatttt ttctcacgag gatggaacta gattgggagt tgaacacttg
     1021 gttgctttgg gacatcaaca gattgctttg ttggctggac cattgtcttc cgtttctgct
     1081 agattgagat tggccggatg gcacaagtac ttgaccagaa accagattca accaattgct
     1141 gagagagagg gagattggtc tgctatgtct ggattccagc agactatgca gatgttgaac
     1201 gaaggaattg tcccaaccgc tatgttggtc gctaatgacc aaatggcttt gggagctatg
     1261 agagctatta ctgaatctgg attgagagtc ggagctgaca tttctgttgt tggatatgat
     1321 gacactgagg attcttcttg ctacattcca ccattgacta ctattaagca agacttcaga
     1381 ttgttgggac agacttctgt tgatagattg ttgcagttgt cccaaggaca agctgttaaa
     1441 ggaaaccaat tgttgccagt ttctttggtt aagagaaaga ctactttggc tccaaacact
     1501 cagactgctt ccccaagagc tttggctgac tctttgatgc aattggctag acaagtctct
     1561 agattggagt ctggacaagg tggcggcggc tctgttaaca actccatgaa ggatttctta
     1621 ggcaagaaaa cggtggatgg agctgatagt ctcaatttgg ccgtgaatct gcaacaacag
     1681 cagagttcaa acacaattgc caatcaatcg ctttcctcaa ttggattgga aagttttggt
     1741 tacggctctg gtatcaaaaa cgagtttaac ttccaagact tgataggttc aaactctggc
     1801 agttcagatc cgacattttc agtagacgct gacgaggccc aaaaactcga catttccaac
     1861 aagaacagtc gtaagagaca gaaactaggt ttgctgccgg tcagcaatgc aacttcccat
     1921 ttgaacggtt tcaatggaat gtccaatgga aagtcacact ctttctcttc accgtctggg
     1981 actaatgacg atgaactaag tggcttgatg ttcaactcac caagcttcaa ccccctcaca
     2041 gttaacgatt ctaccaacaa cagcaaccac aatataggtt tgtctccgat gtcatgctta
     2101 ttttctacag ttcaagaagc atctcaaaaa aagcatggaa attccagtag acacttttca
     2161 tacccatctg ggccggagga cctttggttc aatgagttcc aaaaacaggc cctcacagcc
     2221 aatggagaaa atgctgtcca acagggagat gatgcttcta agaacaacac agccattcct
     2281 aaggaccagt cttcgaactc atcgattttc agttcacgtt ctagtgcagc ttctagcaac
     2341 tcaggagacg atattggaag gatgggccca ttctccaaag gaccagagat tgagttcaac
     2401 tacgattctt ttttggaatc gttgaaggca gagtcaccct cttcttcaaa gtacaatctg
     2461 ccggaaactt tgaaagagta catgaccctt agttcgtctc atctgaatag tcaacactcc
     2521 gacactttgg caaatggcac taacggtaac tattctagca ccgtttccaa caacttgagc
     2581 ttaagtttga actccttctc tttctctgac aagttctcat tgagtccacc aacaatcact
     2641 gacgccgaaa agttttcatt gatgagaaac ttcattgaca acatctcgcc atggtttgac
     2701 acttttgaca ataccaaaca gtttggaaca aaaattccag ttctggccaa aaaatgttct
     2761 tcattgtact atgccattct ggctatatct tctcgtcaaa gagaaaggat aaagaaagag
     2821 cacaatgaaa aaacattgca atgctaccaa tactcactac aacagctcat ccctactgtt
     2881 caaagctcaa ataatattga gtacattatc acatgtattc tcctgagtgt gttccacatc
     2941 atgtctagtg aaccttcaac ccagagggac atcattgtgt cattggcaaa atacattcaa
     3001 gcatgcaaca taaacggatt tacatctaat gacaaactgg aaaagagtat tttctggaac
     3061 tatgtcaatt tggatttggc tacttgtgca atcggtgaag agtcaatggt cattcctttt
     3121 agctactggg ttaaagagac aactgactac aagaccattc aagatgtgaa gccatttttc
     3181 accaagaaga ctagcacgac aactgacgat gacttggacg atatgtatgc catctacatg
     3241 ctgtacatta gtggtagaat cattaacctg ttgaactgca gagatgcgaa gctcaatttt
     3301 gagcccaagt gggagttttt gtggaatgaa ctcaatgaat gggaattgaa caaacccttg
     3361 acctttcaaa gtattgttca gttcaaggcc aatgacgaat cgcagggcgg atcaactttt
     3421 ccaactgttc tattctccaa ctctcgaagc tgttacagta accagctgta tcatatgagc
     3481 tacatcatct tagtgcagaa taaaccacga ttatacaaaa tcccctttac tacagtttct
     3541 gcttcaatgt catctccatc ggacaacaaa gctgggatgt ctgcttccag cacacctgct
     3601 tcagaccacc acgcttctgg tgatcatttg tctccaagaa gtgtagagcc ctctctttcg
     3661 acaacgttga gccctccgcc taatgcaaac ggtgcaggta acaagttccg ctctacgctc
     3721 tggcatgcca agcagatctg tgggatttct atcaacaaca accacaacag caatctagca
     3781 gccaaagtga actcattgca accattgtgg cacgctggaa agctaattag ttccaagtct
     3841 gaacatacac agttgctgaa actgttgaac aaccttgagt gtgcaacagg ctggcctatg
     3901 aactggaagg gcaaggagtt aattgactac tggaatgttg aagaataact cgagccgcgg
     3961 cggccgccag ctttctagaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg
     4021 accatcatca tcatcatcat tgagttttag ccttagacat gactgttcct cagttcaagt
     4081 tgggcactta cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca
     4141 tttgcctgag agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat
     4201 aggatttttt ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc
     4261 gcagctgatg aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc
     4321 ttggtatttc ccactcctct tcagagtaca gaagattaag tgagaccttc gtttgtgcgg
     4381 atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc
     4441 tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaattttc
     4501 cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa
     4561 gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc
     4621 tttttcttga aatttttttt tttagttttt ttctctttca gtgacctcca ttgatattta
     4681 agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa
     4741 ctttttttac ttcttgttca ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt
     4801 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     4861 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     4921 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     4981 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     5041 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     5101 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     5161 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     5221 gccgaggagc aggactgaca cgtccgacgg cggcccacgg gtcccaggcc tcggagatcc
     5281 gtcccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
     5341 tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     5401 tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct
     5461 tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa
     5521 ggttttggga cgctcgaagg ctttaatttg caagctggag accaacatgt gagcaaaagg
     5581 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5641 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5701 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5761 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5821 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5881 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5941 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6001 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6061 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6121 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6181 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6241 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc atgagatc
//
