LOCUS       sequence-248143-        4732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LacI-Gal4AD on pPICZ B expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4732)
  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 4732)
  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-ICL1-LacI-Gal4AD
FEATURES             Location/Qualifiers
     source          1..4732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2176..2520
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             2521..2538
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2618..2864
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2879..3292
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3299..3346
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3365..3739
                     /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      3805..4052
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3821..3839)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4127..4715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4207..4226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
       61 atcagatctt catctaacac tttgtatagc acatcgtacc agtttatcaa aatccaacaa
      121 gttttcctct gggtactgaa attcgattag taataaacgg gcgcacttgg tgatctcctc
      181 atcctgttca ctagccttct tttcgcttaa cttcatcaac gagatcaact cgagtctgcg
      241 agcagagaac tgtttccaaa gagtttcatc gctgaaattc agcttctttc gaataaaatg
      301 tgtcactccc actttattac tcgtggtgtt atttgttttc cacgaactag ttgaatcgcc
      361 atctttacta ccgtcctggg tgacatccgg cgaattcggg acaaatgtgc tgttccggta
      421 gcttgtagga agcggcatcc gtagggcaat atacgactat agcttctaaa gcgtagtaca
      481 atgaaatgtt cgaaggaaca acaaacggat ttgtttttcg taggctcaac ccgttgaggt
      541 gtaactcttt agcgaaaggg taagattgat tgttcgaagt agggcctcaa agggaaagag
      601 aaaaaaaaaa atacaccaag agttacgtaa gcatatattt tttacgtaaa gcatgattga
      661 atttcagcag tattgtttaa caaggctgat gtcgtgtgcc aatcaaaaca aaagagattc
      721 gcataatgcc ataattgggg tgtgtgggcg ccccctaaaa cgtctttctc atcatcatct
      781 gcaaccccca tcgaacctca ttaaatcaca tgacttgtgc gatcctcggt caactcgttc
      841 cgtgcaccca ttccaccccg ggctgaccaa cgcaaggttc tccgagagtc cgctacccca
      901 gatttatatc agcaaccagt cacctttttc cgggcacgac tctatatgcc ctggaaaacc
      961 ggagacgatg agcctgactg taaaaggtga cagaaccccc aactctggtt aatctcttca
     1021 acaaatactt tattttcttt caattcaaag aacacagtat caagtatatc aagaatgggt
     1081 gttaagccag ttactttgta tgacgttgct gaatacgctg gagtttccta ccaaactgtc
     1141 tctagagttg ttaatcaagc ttctcatgtc tccgctaaga ctagagagaa ggttgaggct
     1201 gctatggctg aattgaacta tattccaaat agagttgctc agcagttggc tggaaagcaa
     1261 tctttgttga ttggagtcgc tacttcttct ttggctttgc atgctccatc tcagattgtt
     1321 gctgctatta agtccagagc tgaccagttg ggagcttctg ttgttgtttc tatggttgag
     1381 agatctggag ttgaggcttg caaggctgct gttcataact tgttggctca gagagtttct
     1441 ggattgatta ttaattaccc attggacgat caagacgcta ttgccgttga ggccgcttgt
     1501 accaacgtcc cagctttgtt cttggacgtt tccgatcaaa ctccaattaa ttctattatt
     1561 ttttctcacg aggatggaac tagattggga gttgaacact tggttgcttt gggacatcaa
     1621 cagattgctt tgttggctgg accattgtct tccgtttctg ctagattgag attggccgga
     1681 tggcacaagt acttgaccag aaaccagatt caaccaattg ctgagagaga gggagattgg
     1741 tctgctatgt ctggattcca gcagactatg cagatgttga acgaaggaat tgtcccaacc
     1801 gctatgttgg tcgctaatga ccaaatggct ttgggagcta tgagagctat tactgaatct
     1861 ggattgagag tcggagctga catttctgtt gttggatatg atgacactga ggattcttct
     1921 tgctacattc caccattgac tactattaag caagacttca gattgttggg acagacttct
     1981 gttgatagat tgttgcagtt gtcccaagga caagctgtta aaggaaacca attgttgcca
     2041 gtttctttgg ttaagagaaa gactactttg gctccaaaca ctcagactgc ttccccaaga
     2101 gctttggctg actctttgat gcaattggct agacaagtct ctagattgga gtctggacaa
     2161 ggtggcggcg gctctgccaa ttttaatcaa agtgggaata ttgctgatag ctcattgtcc
     2221 ttcactttca ctaacagtag caacggtccg aacctcataa caactcaaac aaattctcaa
     2281 gcgctttcac aaccaattgc ctcctctaac gttcatgata acttcatgaa taatgaaatc
     2341 acggctagta aaattgatga tggtaataat tcaaaaccac tgtcacctgg ttggacggac
     2401 caaactgcgt ataacgcgtt tggaatcact acagggatgt ttaataccac tacaatggat
     2461 gatgtatata actatctatt cgatgatgaa gataccccac caaacccaaa aaaagagtaa
     2521 catcatcatc atcatcattg agtttgtagc cttagacatg actgttcctc agttcaagtt
     2581 gggcacttac gagaagaccg gtcttgctag attctaatca agaggatgtc agaatgccat
     2641 ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata
     2701 ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg
     2761 cagctgatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct
     2821 tggtatttcc cactcctctt cagagtacag aagattaagt gagaccttcg tttgtgcgga
     2881 tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc cagattttct
     2941 cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc
     3001 ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag
     3061 agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct
     3121 ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc attgatattt
     3181 aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca
     3241 acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta agggcggtgt
     3301 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     3361 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     3421 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     3481 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     3541 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     3601 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     3661 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     3721 ggccgaggag caggactgac acgtccgacg cggcccgacg ggtccgaggc ctcggagatc
     3781 cgtccccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc
     3841 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     3901 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
     3961 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
     4021 aggttttggg acgctcgaag gctttaattt gcaagctgga gaccaacatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4561 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc tt
//
