LOCUS       sequence-248174-        4624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LacI-VP16 on pPICZ B expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4624)
  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 4624)
  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-VP16
FEATURES             Location/Qualifiers
     source          1..4624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2411..2428
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2508..2754
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2769..3182
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3189..3236
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3255..3629
                     /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      3695..3942
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3711..3729)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4017..4605)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4097..4116)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
       61 cagatcttca tctaacactt tgtatagcac atcgtaccag tttatcaaaa tccaacaagt
      121 tttcctctgg gtactgaaat tcgattagta ataaacgggc gcacttggtg atctcctcat
      181 cctgttcact agccttcttt tcgcttaact tcatcaacga gatcaactcg agtctgcgag
      241 cagagaactg tttccaaaga gtttcatcgc tgaaattcag cttctttcga ataaaatgtg
      301 tcactcccac tttattactc gtggtgttat ttgttttcca cgaactagtt gaatcgccat
      361 ctttactacc gtcctgggtg acatccggcg aattcgggac aaatgtgctg ttccggtagc
      421 ttgtaggaag cggcatccgt agggcaatat acgactatag cttctaaagc gtagtacaat
      481 gaaatgttcg aaggaacaac aaacggattt gtttttcgta ggctcaaccc gttgaggtgt
      541 aactctttag cgaaagggta agattgattg ttcgaagtag ggcctcaaag ggaaagagaa
      601 aaaaaaaaat acaccaagag ttacgtaagc atatattttt tacgtaaagc atgattgaat
      661 ttcagcagta ttgtttaaca aggctgatgt cgtgtgccaa tcaaaacaaa agagattcgc
      721 ataatgccat aattggggtg tgtgggcgcc ccctaaaacg tctttctcat catcatctgc
      781 aacccccatc gaacctcatt aaatcacatg acttgtgcga tcctcggtca actcgttccg
      841 tgcacccatt ccaccccggg ctgaccaacg caaggttctc cgagagtccg ctaccccaga
      901 tttatatcag caaccagtca cctttttccg ggcacgactc tatatgccct ggaaaaccgg
      961 agacgatgag cctgactgta aaaggtgaca gaacccccaa ctctggttaa tctcttcaac
     1021 aaatacttta ttttctttca attcaaagaa cacagtatca agtatatcaa gaatgggtgt
     1081 taagccagtt actttgtatg acgttgctga atacgctgga gtttcctacc aaactgtctc
     1141 tagagttgtt aatcaagctt ctcatgtctc cgctaagact agagagaagg ttgaggctgc
     1201 tatggctgaa ttgaactata ttccaaatag agttgctcag cagttggctg gaaagcaatc
     1261 tttgttgatt ggagtcgcta cttcttcttt ggctttgcat gctccatctc agattgttgc
     1321 tgctattaag tccagagctg accagttggg agcttctgtt gttgtttcta tggttgagag
     1381 atctggagtt gaggcttgca aggctgctgt tcataacttg ttggctcaga gagtttctgg
     1441 attgattatt aattacccat tggacgatca agacgctatt gccgttgagg ccgcttgtac
     1501 caacgtccca gctttgttct tggacgtttc cgatcaaact ccaattaatt ctattatttt
     1561 ttctcacgag gatggaacta gattgggagt tgaacacttg gttgctttgg gacatcaaca
     1621 gattgctttg ttggctggac cattgtcttc cgtttctgct agattgagat tggccggatg
     1681 gcacaagtac ttgaccagaa accagattca accaattgct gagagagagg gagattggtc
     1741 tgctatgtct ggattccagc agactatgca gatgttgaac gaaggaattg tcccaaccgc
     1801 tatgttggtc gctaatgacc aaatggcttt gggagctatg agagctatta ctgaatctgg
     1861 attgagagtc ggagctgaca tttctgttgt tggatatgat gacactgagg attcttcttg
     1921 ctacattcca ccattgacta ctattaagca agacttcaga ttgttgggac agacttctgt
     1981 tgatagattg ttgcagttgt cccaaggaca agctgttaaa ggaaaccaat tgttgccagt
     2041 ttctttggtt aagagaaaga ctactttggc tccaaacact cagactgctt ccccaagagc
     2101 tttggctgac tctttgatgc aattggctag acaagtctct agattggagt ctggacaagg
     2161 tggcggcggc tctgctccac caaccgacgt ttctttgggt gacgagttgc acttggacgg
     2221 tgaagatgtt gccatggctc atgctgacgc tttggacgac ttcgacttgg acatgttggg
     2281 tgacggtgat tctccaggtc caggtttcac tccacacgat tctgctccat acggtgcttt
     2341 ggacatggcc gacttcgagt ttgagcagat gttcaccgac gctttgggta ttgacgagta
     2401 cggtggttaa catcatcatc atcatcattg agtttgtagc cttagacatg actgttcctc
     2461 agttcaagtt gggcacttac gagaagaccg gtcttgctag attctaatca agaggatgtc
     2521 agaatgccat ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct
     2581 atatagtata ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca
     2641 gcctatctcg cagctgatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg
     2701 atgtttttct tggtatttcc cactcctctt cagagtacag aagattaagt gagaccttcg
     2761 tttgtgcgga tcccccacac accatagctt caaaatgttt ctactccttt tttactcttc
     2821 cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag cacagcatac
     2881 taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa aggtttggaa
     2941 aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata aaaattttta
     3001 tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
     3061 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
     3121 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
     3181 agggcggtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3241 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     3301 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3361 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3421 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     3481 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     3541 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     3601 tgcacttcgt ggccgaggag caggactgac acgtccgacg cggcccgacg ggtccgaggc
     3661 ctcggagatc cgtccccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
     3721 cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     3781 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     3841 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     3901 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcaagctgga gaccaacatg
     3961 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4021 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4081 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4141 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4201 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4261 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4321 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4381 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4441 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4501 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4561 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4621 tttt
//
