LOCUS       sequence-248145-        3527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid overexpressing NpgA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3527)
  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 3527)
  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_lacO-cP-AOX1-NpgA
FEATURES             Location/Qualifiers
     source          1..3527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    73..89
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             837..845
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1310..1327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1407..1653
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1668..2081
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2088..2135
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2154..2528
                     /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      2594..2841
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2610..2628)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2916..3504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2996..3015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagatcaga
       61 tcttgtgtgg aattgtgagc ggataacaat ttcacacact aacccctact tgacagcaat
      121 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      181 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      241 caacttgaga agatcaaaaa acaactaatt attcgaaatg gtgcaagaca catcaagcgc
      301 aagcacttcg ccaattttaa caagatggta catcgacacc cgccctctaa ccgcctcaac
      361 agcagccctt cctctccttg aaaccctcca gcccgctgat caaatctccg tccaaaaata
      421 ctaccatctg aaggataaac acatgtctct cgcctctaat ctgctcaaat acctcttcgt
      481 ccaccgaaac tgtcgcatcc cctggtcttc aatcgtgatc tctcgaaccc cagatccgca
      541 cagacgacca tgctatattc caccctcagg ctcacaggaa gacagcttca aagacggata
      601 taccggcatc aacgttgagt tcaacgtcag ccaccaagcc tcaatggtcg cgatcgcggg
      661 aacagctttt actcccaata gtggtgggga cagcaaactc aaacccgaag tcggaattga
      721 tattacgtgc gtaaacgagc ggcagggacg gaacggggaa gagcggagcc tggaatcgct
      781 acgtcaatat attgatatat tctcggaagt gttttccact gcagagatgg ccaatataag
      841 gaggttagat ggagtctcat catcctcact gtctgctgat cgtcttgtgg actacgggta
      901 cagactcttc tacacttact gggcgctcaa agaggcgtat ataaaaatga ctggggaggc
      961 cctcttagca ccgtggttac gggaactgga attcagtaat gtcgtcgccc cggccgctgt
     1021 tgcggagagt ggggattcgg ctggggattt cggggagccg tatacgggtg tcaggacgac
     1081 tttatataaa aatctcgttg aggatgtgag gattgaagtt gctgctctgg gcggtgatta
     1141 cctatttgca acggctgcga ggggtggtgg gattggagct agttctagac caggaggtgg
     1201 tccagacgga agtggtatcc gaagccagga tccctggagg cctttcaaga agttagatat
     1261 agagcgagat atccagccct gtgcgactgg ggtgtgtaat tgcctatccc atcatcatca
     1321 tcatcattga gtttgtagcc ttagacatga ctgttcctca gttcaagttg ggcacttacg
     1381 agaagaccgg tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag
     1441 atgcaggctt catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt
     1501 gtcattttgt ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa
     1561 tatcttgtgg taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc
     1621 actcctcttc agagtacaga agattaagtg agaccttcgt ttgtgcggat cccccacaca
     1681 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
     1741 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
     1801 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
     1861 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
     1921 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa
     1981 cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac
     2041 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gggcggtgtt gacaattaat
     2101 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     2161 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     2221 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     2281 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     2341 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     2401 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     2461 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     2521 aggactgaca cgtccgacgc ggcccgacgg gtccgaggcc tcggagatcc gtcccccttt
     2581 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca
     2641 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     2701 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2761 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
     2821 cgctcgaagg ctttaatttg caagctggag accaacatgt gagcaaaagg ccagcaaaag
     2881 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2941 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3001 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3061 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctac
//
