LOCUS       sequence-252455-        5219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Z. rouxii episomal vector containing ClonNAT resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5219)
  AUTHORS   Bizzarri M, Cassanelli S, Duskova M, Sychrova H, Solieri L
  TITLE     A set of plasmids carrying antibiotic resistance markers and Cre 
            recombinase for genetic engineering of nonconventional yeast 
            Zygosaccharomyces rouxii.
  JOURNAL   Yeast. 2019 Dec;36(12):711-722. doi: 10.1002/yea.3438. Epub 2019 Sep
            11.
  PUBMED    31414502
REFERENCE   2  (bases 1 to 5219)
  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"; doi: "10.1002/yea.3438"; 
            journalName: "Yeast"; date: "2019-12"; volume: "36"; issue: "12"; 
            pages: "711-722"
COMMENT     Sequence Label: pZEN
FEATURES             Location/Qualifiers
     source          1..5219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(146..469)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    396..452
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(465..481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(465..481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(478..500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    489..505
                     /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)."
     promoter        complement(join(513..519,520..537,538..543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    558..579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        complement(1931..2142)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(2220..2237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3400..3988)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3480..3499)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4159..4950,4951..5019))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4782..4801
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5020..5124)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5192..5210
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      601 gctttccagt cgggaaacct gtcgtgccag cagatctggc tttttctttc caattttttt
      661 tttgtgagtt tagtatacat gcatttactt ataatacagt tttatggacg gtggtatgtt
      721 ttagtttagc ttcaattcta attgattgat taatcagttg attggtttca atatgacaaa
      781 tgggtagggt gggaaaactt cattttcaat tcagatcaaa cttttttgtt gtcgacataa
      841 tatttctcgt ttgggatgtt actgtcacat taataataca cacacatcag cttataattt
      901 tgaaagtaat ttatcagata tgttgtgacg atcagtggaa atggctaact tcaatgtatc
      961 tgttcttccc ctttttcaaa gttcacgttt tttgattgat tgattgatct gtcggcagtg
     1021 gtttcaaaac cattcggtga gtaatcctat caatcaatgt tacgacaaaa ggctcaatat
     1081 tcaaaattgc aatgttttat gttttcctac gtgtacttgt gcaaggcaat tgattcaaca
     1141 ttgcttttgg tgtttgacga gtttctagtt tggacttgtg ttgttatctg ggctatacag
     1201 atttcccggc tcactatgaa tttttttttt cgacgctcag tgcacacaac tataaacaac
     1261 acaaacacaa acacagcaag aaaaaaaaaa aacgaacatt gaattgaaac caagccaact
     1321 gaaaaattcc ttatttaaat gactgtcata ctaacccatt tttatagaag aagttgctgc
     1381 tttagttatc gataacggtt ctcatatgaa aatttcggtg atccctgagc aggtggcgga
     1441 aacattggat gctgagaacc atttcattgt tcgtgaagtg ttcgatgtgc acctatccga
     1501 ccaaggcttt gaactatcta ccagaagtgt gagcccctac cggaaggatt acatctcgga
     1561 tgatgactct gatgaagact ctgcttgcta tggcgcattc atcgaccaag agcttgtcgg
     1621 gaagattgaa ctcaactcaa catggaacga tctagcctct atcgaacaca ttgttgtgtc
     1681 gcacacgcac cgaggcaaag gagtcgcgca cagtctcatc gaatttgcga aaaagtgggc
     1741 actaagcaga cagctccttg gcatacgatt agagacacaa acgaacaatg tacctgcctg
     1801 caatttgtac gcaaaatgtg gctttactct cggcggcatt gacctcttca cgtataaaac
     1861 tagacctcaa gtctcgaacg aaacagcgat gtactggtac tggttctcgg gagcacagga
     1921 tgacgcctaa gatttatctt cgtttccggt ttttgttctg tgcagttggg ttaagaatac
     1981 tgggcaattt catgtttctt caacactaca tatgcgtata tataccaatc taagtctgtg
     2041 ctccttcctt cgttcttcct tctgttcgga gattaccgaa tcaaaaaaat ttcaaggaaa
     2101 ccgaaatcaa aaaaaagaat aaaaaaaaaa tgatgaattg aaaagccaga tctgctgcat
     2161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2221 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agcatcgata
     2281 gatcctacaa aatatgtaat gctattctta gattcccacg agaggcttgc ttttgtacat
     2341 agaccctcca gaaaacatag tgctgtttat gcagcacaca gaaggtttat aggtgtgagc
     2401 tacactttat aggccgggta agtatagtaa gatatgtgca aattagccgc ggagccataa
     2461 acaatcatct tttaaattag atggcacagc cttaaaccaa gcaattctgg tcacctctcc
     2521 ggattttggg tgaatatcaa aatgacctca caaggtaaca ctttggatga tattcggtta
     2581 caagtcaggt gtgacattct aggtaaagac accaacagaa gaggaggctc tgagaaggat
     2641 cctgaggatc aaagtgaaac tgagtccagt ggatagactc atttgctaat ttaagccttt
     2701 ctgtaagact attgagaccg tgtactatta atttttctta ttgtaacagt aggtaatagg
     2761 taagggtata aaaatcatgg gtataatgat aaatttgtat tgcaataatg tccaaatcaa
     2821 attcggaggc gaggtccaag aagcaagatg gtgtatggta gcaatataga gagggtttta
     2881 ttgggttatt attttatttt caagaattta cattttgagt gtagatcttt actttcagca
     2941 agaaggtacg aggaacaagc gggtagttta gaggaacaaa ataaaaatac aagagcatag
     3001 taactgtcag ggtgtatcaa ctggaccaat cattgcttct cgtagtcaaa tgtttattcg
     3061 atagagtaat ggtttaattt ctgtagcaac cgcctgcgca ggctcccttc tgcgcggtac
     3121 gggttgatga aagaacgact tgatgaaaga acgatttgat gaaagaacga attaatgaaa
     3181 gaacgatttg atgaaagaat aacgtattct ttcatcaacg tccacagtga tatgatctcg
     3241 cctagaaaaa ctaaattata ttaagctaaa ctaaatcgat gctcactcaa aggcggtaat
     3301 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3361 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3421 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3481 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3541 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3601 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3661 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3721 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3781 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3841 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3901 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3961 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4021 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4081 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4141 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4201 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4261 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4321 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4381 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4441 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4501 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4561 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4621 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4681 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4741 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4801 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4861 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4921 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4981 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5041 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5101 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     5161 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
