LOCUS       sequence-249046-        4580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast plasmid overexpressing CPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4580)
  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 4580)
  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-CPR
FEATURES             Location/Qualifiers
     source          1..4580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        384..431
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             450..824
                     /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      890..1137
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(906..924)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(1212..1800)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1292..1311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1896..1912
                     /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)."
     CDS             4186..4203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4283..4529
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        join(4544..4580,1..377)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccttttttac tcttccagat tttctcggac tccgcgcatc gccgtaccac ttcaaaacac
       61 ccaagcacag catactaaat ttcccctctt tcttcctcta gggtgtcgtt aattacccgt
      121 actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg
      181 caataaaaat ttttatcacg tttctttttc ttgaaaattt ttttttttga tttttttctc
      241 tttcgatgac ctcccattga tatttaagtt aataaacggt cttcaatttc tcaagtttca
      301 gtttcatttt tcttgttcta ttacaacttt ttttacttct tgctcattag aaagaaagca
      361 tagcaatcta atctaagggc ggtgttgaca attaatcatc ggcatagtat atcggcatag
      421 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
      481 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
      541 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
      601 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
      661 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
      721 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
      781 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtc cgacgcggcc
      841 cgacgggtcc gaggcctcgg agatccgtcc cccttttcct ttgtcgatat catgtaatta
      901 gttatgtcac gcttacattc acgccctccc cccacatccg ctctaaccga aaaggaagga
      961 gttagacaac ctgaagtcta ggtccctatt tattttttta tagttatgtt agtattaaga
     1021 acgttattta tatttcaaat ttttcttttt tttctgtaca gacgcgtgta cgcatgtaac
     1081 attatactga aaaccttgct tgagaaggtt ttgggacgct cgaaggcttt aatttgcaag
     1141 ctggagacca acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1621 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1681 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1741 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1861 gggattttgg tcatgagatc agatcttgtg tggaattgtg agcggataac aatttcacac
     1921 actaacccct acttgacagc aatatataaa cagaaggaag ctgccctgtc ttaaaccttt
     1981 ttttttatca tcattattag cttactttca taattgcgac tggttccaat tgacaagctt
     2041 ttgattttaa cgacttttaa cgacaacttg agaagatcaa aaaacaacta attattcgaa
     2101 atggctcaac tcgacactct cgacctggtg gtcctggtgg tgcttttggt gggtagcgcc
     2161 gcctacttca ccaagggcac ctactgggcc gttcccaagg acccgtatgc cgcctccggt
     2221 cccgccatga atggtggcgc caaggcgggc aaatccaggg acatcattga gaaaatggaa
     2281 gagactggca agaactgtgt gattttctac ggctcgcaga ccggtaccgc cgaggattat
     2341 gcgtcgcgcc tggccaagga aggctcccag cgtttcggcc tcaagaccat ggtcgcagat
     2401 ctggaagact acgattatga gaacctggac aagttccccg aggacaaggt tgccttcttc
     2461 gtcatggcca cctatggtga gggtgaaccc accgacaacg ccgtcgagtt ctaccagttc
     2521 atctcgggtg aggacgtcgc gttcgagagc ggcgcctctg ccgacgacaa gcccctgtcc
     2581 tccctcaagt atgtcacttt cggtctcggt aacaacacct atgagcacta tcaggctatg
     2641 gttcgcaatc tggatgccgc tctcaccaag ctgggtgcgc agcgcattgg agatgctggt
     2701 gaaggtgatg acggcgctgg caccatggaa gaagatttcc tggcctggaa agagcccatg
     2761 tggactgccc tgtccgaggc catgaacctt caggagcgcg aagccgtcta tgagccggtg
     2821 ttctcggtca cggaagatga atccctgtcc cccgaagacg aagccgtcta cctcggtgag
     2881 ccgaccaagg gtcaccgtga cggcaccccc agtggcccgt attccgctca caaccccttc
     2941 atcgccccca tcgtcgagtc tcgtgaactg ttcaacgtca aggaccgtaa ttgtctgcac
     3001 atggagatca gcatcgctgg tagcaacctt tcttaccaga ctggtgatca catcgcgatt
     3061 tggcccacga acgctggtgc cgaggtggac cggttcctcc aggtgtttgg tcttgagaac
     3121 aagcgtcatt ccgtcatcaa catcaagggt atcgatgtga ccgccaaggt tcccattccg
     3181 actcccacca cgtatgatgc tgctgttcgc tactatatgg aaattgctgc gcccgtctcc
     3241 cgtcagtttg tggctaccct ggctgcgttt gctcccgatg aggagactaa ggcggaaatc
     3301 gtgcgtttgg gtagcgacaa ggactacttc cacgagaaaa tcagcaacca gtgcttcacc
     3361 atcgctcagg ctcttcagag tgtcacctcc aagcccttct cggctgtccc gttctctctg
     3421 cttatcgagg gtctcaataa gctccagccc cgttactact ccatctcttc ctcctccatg
     3481 gtccagaagg ataagatcag cattactgcc gtcgtggaat ccactcgctt gcctggtgcc
     3541 gcccaccttg tcaagggtgt cacgaccaac tatctccttg ccctgaagca aaagcagaat
     3601 ggggatccgt ctcccgaccc tcacggctta acttatacta tcactgggcc ccgtaacaag
     3661 tacgacggaa tccacgttcc cgttcacgtc cgccactcca atttcaagct cccctctgat
     3721 ccctctcggc ccattatcat ggttggccct ggtaccggtg tggctccctt ccgtggattc
     3781 atccaggagc gtgccgcctt ggccgccaag ggtgagaacg tcggtcccac cgtgttgttc
     3841 tttggatgcc gcaggcgcga tgaggacttt atgtacgcag atgaattcaa gacctaccag
     3901 gaacagcttg gggacaagct tcagatcatt actgcgtttt ctcgtgaaac ttcccagaag
     3961 gtgtatgttc agcacagact gcgtgaacac tccgatctgg tgagcagcct cctgaagcag
     4021 aaggctaact tttacgtctg cggtgacgcc gccaacatgg cgcgtgaagt caaccttgtg
     4081 cttggccaga tcatcgcgca acagcggggt ctcccggctg aacgggccga ggaaatggtg
     4141 aagcacatgc gcagcagcgg cagctaccag gaggacgtgt ggtcacatca tcatcatcat
     4201 cattgagttt gtagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa
     4261 gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc
     4321 aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca
     4381 ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc
     4441 ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc
     4501 ctcttcagag tacagaagat taagtgagac cttcgtttgt gcggatcccc cacacaccat
     4561 agcttcaaaa tgtttctact
//
