LOCUS       sequence-248156-        6213 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 6213)
  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 6213)
  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..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        38..513
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     CDS             3970..3987
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4067..4313
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        4328..4741
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        4748..4795
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4814..5188
                     /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      5254..5501
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5270..5288)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(5576..6164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5656..5675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actcacgtta agggattttg gtcatgagat cagatctttt ttgtagaaat gtcttggtgt
       61 cctcgaccaa tcaggtagcc atccctgaaa tacctggctc cgtggcaaca ccgaacgacc
      121 tgctggcaac gttaaattct ccggggtaaa acttaaatgt ggagtaatag aaccagaaac
      181 gtctcttccc ttctctctcc ttccaccgcc cgttaccgtc cctaggaaat tttactctgc
      241 tggagagctt cttctacggc ccccttgcag caatgctctt cccagcatta cgttgcgggt
      301 aaaacggagg tcgtgtaccc gacctagcag cccagggatg gaaagtcccg gccgtcgctg
      361 gcaataactg cgggcggacg catgtcttga gattattgga aaccaccaga atcgaatata
      421 aaaggcgaac acctttccca attttggttt ctcctgaccc aaagacttta aatttaattt
      481 atttgtccct atttcaatca attgaacaac tatatgggtg ttaagccagt tactttgtat
      541 gacgttgctg aatacgctgg agtttcctac caaactgtct ctagagttgt taatcaagct
      601 tctcatgtct ccgctaagac tagagagaag gttgaggctg ctatggctga attgaactat
      661 attccaaata gagttgctca gcagttggct ggaaagcaat ctttgttgat tggagtcgct
      721 acttcttctt tggctttgca tgctccatct cagattgttg ctgctattaa gtccagagct
      781 gaccagttgg gagcttctgt tgttgtttct atggttgaga gatctggagt tgaggcttgc
      841 aaggctgctg ttcataactt gttggctcag agagtttctg gattgattat taattaccca
      901 ttggacgatc aagacgctat tgccgttgag gccgcttgta ccaacgtccc agctttgttc
      961 ttggacgttt ccgatcaaac tccaattaat tctattattt tttctcacga ggatggaact
     1021 agattgggag ttgaacactt ggttgctttg ggacatcaac agattgcttt gttggctgga
     1081 ccattgtctt ccgtttctgc tagattgaga ttggccggat ggcacaagta cttgaccaga
     1141 aaccagattc aaccaattgc tgagagagag ggagattggt ctgctatgtc tggattccag
     1201 cagactatgc agatgttgaa cgaaggaatt gtcccaaccg ctatgttggt cgctaatgac
     1261 caaatggctt tgggagctat gagagctatt actgaatctg gattgagagt cggagctgac
     1321 atttctgttg ttggatatga tgacactgag gattcttctt gctacattcc accattgact
     1381 actattaagc aagacttcag attgttggga cagacttctg ttgatagatt gttgcagttg
     1441 tcccaaggac aagctgttaa aggaaaccaa ttgttgccag tttctttggt taagagaaag
     1501 actactttgg ctccaaacac tcagactgct tccccaagag ctttggctga ctctttgatg
     1561 caattggcta gacaagtctc tagattggag tctggacaag gtggcggcgg ctctgttaac
     1621 aactccatga aggatttctt aggcaagaaa acggtggatg gagctgatag tctcaatttg
     1681 gccgtgaatc tgcaacaaca gcagagttca aacacaattg ccaatcaatc gctttcctca
     1741 attggattgg aaagttttgg ttacggctct ggtatcaaaa acgagtttaa cttccaagac
     1801 ttgataggtt caaactctgg cagttcagat ccgacatttt cagtagacgc tgacgaggcc
     1861 caaaaactcg acatttccaa caagaacagt cgtaagagac agaaactagg tttgctgccg
     1921 gtcagcaatg caacttccca tttgaacggt ttcaatggaa tgtccaatgg aaagtcacac
     1981 tctttctctt caccgtctgg gactaatgac gatgaactaa gtggcttgat gttcaactca
     2041 ccaagcttca accccctcac agttaacgat tctaccaaca acagcaacca caatataggt
     2101 ttgtctccga tgtcatgctt attttctaca gttcaagaag catctcaaaa aaagcatgga
     2161 aattccagta gacacttttc atacccatct gggccggagg acctttggtt caatgagttc
     2221 caaaaacagg ccctcacagc caatggagaa aatgctgtcc aacagggaga tgatgcttct
     2281 aagaacaaca cagccattcc taaggaccag tcttcgaact catcgatttt cagttcacgt
     2341 tctagtgcag cttctagcaa ctcaggagac gatattggaa ggatgggccc attctccaaa
     2401 ggaccagaga ttgagttcaa ctacgattct tttttggaat cgttgaaggc agagtcaccc
     2461 tcttcttcaa agtacaatct gccggaaact ttgaaagagt acatgaccct tagttcgtct
     2521 catctgaata gtcaacactc cgacactttg gcaaatggca ctaacggtaa ctattctagc
     2581 accgtttcca acaacttgag cttaagtttg aactccttct ctttctctga caagttctca
     2641 ttgagtccac caacaatcac tgacgccgaa aagttttcat tgatgagaaa cttcattgac
     2701 aacatctcgc catggtttga cacttttgac aataccaaac agtttggaac aaaaattcca
     2761 gttctggcca aaaaatgttc ttcattgtac tatgccattc tggctatatc ttctcgtcaa
     2821 agagaaagga taaagaaaga gcacaatgaa aaaacattgc aatgctacca atactcacta
     2881 caacagctca tccctactgt tcaaagctca aataatattg agtacattat cacatgtatt
     2941 ctcctgagtg tgttccacat catgtctagt gaaccttcaa cccagaggga catcattgtg
     3001 tcattggcaa aatacattca agcatgcaac ataaacggat ttacatctaa tgacaaactg
     3061 gaaaagagta ttttctggaa ctatgtcaat ttggatttgg ctacttgtgc aatcggtgaa
     3121 gagtcaatgg tcattccttt tagctactgg gttaaagaga caactgacta caagaccatt
     3181 caagatgtga agccattttt caccaagaag actagcacga caactgacga tgacttggac
     3241 gatatgtatg ccatctacat gctgtacatt agtggtagaa tcattaacct gttgaactgc
     3301 agagatgcga agctcaattt tgagcccaag tgggagtttt tgtggaatga actcaatgaa
     3361 tgggaattga acaaaccctt gacctttcaa agtattgttc agttcaaggc caatgacgaa
     3421 tcgcagggcg gatcaacttt tccaactgtt ctattctcca actctcgaag ctgttacagt
     3481 aaccagctgt atcatatgag ctacatcatc ttagtgcaga ataaaccacg attatacaaa
     3541 atccccttta ctacagtttc tgcttcaatg tcatctccat cggacaacaa agctgggatg
     3601 tctgcttcca gcacacctgc ttcagaccac cacgcttctg gtgatcattt gtctccaaga
     3661 agtgtagagc cctctctttc gacaacgttg agccctccgc ctaatgcaaa cggtgcaggt
     3721 aacaagttcc gctctacgct ctggcatgcc aagcagatct gtgggatttc tatcaacaac
     3781 aaccacaaca gcaatctagc agccaaagtg aactcattgc aaccattgtg gcacgctgga
     3841 aagctaatta gttccaagtc tgaacataca cagttgctga aactgttgaa caaccttgag
     3901 tgtgcaacag gctggcctat gaactggaag ggcaaggagt taattgacta ctggaatgtt
     3961 gaagaataac atcatcatca tcatcattga gtttgtagcc ttagacatga ctgttcctca
     4021 gttcaagttg ggcacttacg agaagaccgg tcttgctaga ttctaatcaa gaggatgtca
     4081 gaatgccatt tgcctgagag atgcaggctt catttttgat acttttttat ttgtaaccta
     4141 tatagtatag gatttttttt gtcattttgt ttcttctcgt acgagcttgc tcctgatcag
     4201 cctatctcgc agctgatgaa tatcttgtgg taggggtttg ggaaaatcat tcgagtttga
     4261 tgtttttctt ggtatttccc actcctcttc agagtacaga agattaagtg agaccttcgt
     4321 ttgtgcggat cccccacaca ccatagcttc aaaatgtttc tactcctttt ttactcttcc
     4381 agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc acagcatact
     4441 aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa ggtttggaaa
     4501 agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa aaatttttat
     4561 cacgtttctt tttcttgaaa attttttttt ttgatttttt tctctttcga tgacctccca
     4621 ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca tttttcttgt
     4681 tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa tctaatctaa
     4741 gggcggtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4801 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     4861 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     4921 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     4981 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     5041 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     5101 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     5161 gcacttcgtg gccgaggagc aggactgaca cgtccgacgc ggcccgacgg gtccgaggcc
     5221 tcggagatcc gtcccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac
     5281 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag
     5341 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc
     5401 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct
     5461 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctggag accaacatgt
     5521 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5761 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6001 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6181 tttctacggg gtctgacgct cagtggaacg aaa
//
