LOCUS       sequence-252438-        5597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Z. rouxii centromeric vector containing ClonNAT resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  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 5597)
  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: pZCAN
FEATURES             Location/Qualifiers
     source          1..5597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             216..521
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIIAGPFS"
     misc_feature    233..289
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(290..307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     RBS             1261..1269
                     /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)"
     promoter        complement(3365..3580)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3605..3623)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3691..3795
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3796..3864,3865..4656)
                     /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     complement(4014..4033)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4827..5415
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5316..5335
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5569..5586
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
      481 gcggtatttc acaccgcata atcgctgggc cattctcatg aagaatatct tgaatttatt
      541 gtcatattac tagtaggcat ttgtttatta ccagcttacg atataaagtc gttcatcttc
      601 tttatggcga gaaactgtaa ttgcgaaaat tgtctaggca atttttttca gtagttagtc
      661 tgagattttt ccttgtgaac caaccttcct gtcgttatta atactagaag ataagacgaa
      721 acagtttagt ttgtaggtac gtaaaaactt gttcaaagta ggcttcagta atcttgttcc
      781 gaaaaagttt ctcacatatc aaacaaatta gaggtttcgg aagtaagcca caacaaaaat
      841 aatgaatatt ttttaaacgc ttttaaaaca gataattatc aaaaaataga attaaagctt
      901 tatttgtatt aattttgaaa gtattaaata aaattatata ctgtcttata atatatatta
      961 ttatatcatg tgattaaatt acacagcttc taaagctctt taatggattg cggtttttac
     1021 atctactcta ctttagatca tactttgtct ttgattaaag tatattaagt ggccatctgt
     1081 gaccagtatc ttttgaaaca atttgtcata atatccatat tccatcatat taaacattta
     1141 cgttttgttc tactttaacc aatttgaaag acattattta aatcaaaaat attttctccg
     1201 agttagctat gaaaaagatt gaatacctca tgtataaaga tatcctctcc ttccaagaat
     1261 acctccttaa gtttcatttc taccgtcagg actcgtgttt tcgccaggag cgataagggg
     1321 acgtgtgatt tctgcatcat cgttattagg tgacactaag ttagtgcccg atcgatgcta
     1381 gcaagaatcg ggtcattgta gcgtatgcgc ctgtgaacat tctcttcaac aagtttgatt
     1441 ccattgcggt gaaatggtaa aagtcaaccc cctgcgatgt atattttcct gtacaatcaa
     1501 tcaaaaagcc aaatgattta gcattatctt tacatcttgt tattttacag attttatgtt
     1561 tagatctttt atgcttgctt ttcaaaaggc ttgcaggcaa gtgcacaaac aatacttaaa
     1621 taaatactac tcagtaataa cctatttctt agcatttttg acgaaatttg ctattttgtt
     1681 agagtctttt acaccatttg tctccacacc tccgcttaca tcaacaccaa taacgccatt
     1741 taatctaagc gcatcaccaa cattttctgg cgtcagtcca ccagctaaca taaaatgtaa
     1801 gctctcgggg ctctcttgcc ttccaaccca gtcagaaatc gagttccaat ccaaaagttc
     1861 acctgtccca cctgcttctg aatcaaacaa gggaataaac gaatgaggtt tctgtgaagc
     1921 tgcactgagt agtatgttgc agtcttttgg aaatacgagt cttttaataa ctggcaaacc
     1981 gaggaactct tggtattctt gccacgactc atctccatgc agttggacga tcgatgataa
     2041 gctgtcaaac atgagaattg ggtaataact gatataatta aattgaagct ctaatttgtg
     2101 agtttagtat acatgcattt acttataata cagttttatg gacggtggta tgttttagtt
     2161 tagcttcaat tctaattgat tgattaatca gttgattggt ttcaatatga caaatgggta
     2221 gggtgggaaa acttcatttt caattcagat caaacttttt tgttgtcgac ataatatttc
     2281 tcgtttggga tgttactgtc acattaataa tacacacaca tcagcttata attttgaaag
     2341 taatttatca gatatgttgt gacgatcagt ggaaatggct aacttcaatg tatctgttct
     2401 tccccttttt caaagttcac gttttttgat tgattgattg atctgtcggc agtggtttca
     2461 aaaccattcg gtgagtaatc ctatcaatca atgttacgac aaaaggctca atattcaaaa
     2521 ttgcaatgtt ttatgttttc ctacgtgtac ttgtgcaagg caattgattc aacattgctt
     2581 ttggtgtttg acgagtttct agtttggact tgtgttgtta tctgggctat acagatttcc
     2641 cggctcacta tgaatttttt ttttcgacgc tcagtgcaca caactataaa caacacaaac
     2701 acaaacacag caagaaaaaa aaaaaacgaa cattgaattg aaaccaagcc aactgaaaaa
     2761 ttccttattt aaatgactgt catactaacc catttttata gaagaagttg ctgctttagt
     2821 tatcgataac ggttctcata tgaaaatttc ggtgatccct gagcaggtgg cggaaacatt
     2881 ggatgctgag aaccatttca ttgttcgtga agtgttcgat gtgcacctat ccgaccaagg
     2941 ctttgaacta tctaccagaa gtgtgagccc ctaccggaag gattacatct cggatgatga
     3001 ctctgatgaa gactctgctt gctatggcgc attcatcgac caagagcttg tcgggaagat
     3061 tgaactcaac tcaacatgga acgatctagc ctctatcgaa cacattgttg tgtcgcacac
     3121 gcaccgaggc aaaggagtcg cgcacagtct catcgaattt gcgaaaaagt gggcactaag
     3181 cagacagctc cttggcatac gattagagac acaaacgaac aatgtacctg cctgcaattt
     3241 gtacgcaaaa tgtggcttta ctctcggcgg cattgacctc ttcacgtata aaactagacc
     3301 tcaagtctcg aacgaaacag cgatgtactg gtactggttc tcgggagcac aggatgacgc
     3361 ctaagattta tcttcgtttc ctgcatgttt ttgttctgtg cagttgggtt aagaatactg
     3421 ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct
     3481 ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc
     3541 gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aagctaattc ttgaagacga
     3601 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3661 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3721 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3781 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3841 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3901 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3961 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4021 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4081 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4141 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4201 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4261 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4321 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4381 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4441 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4501 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4561 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4621 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4681 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4741 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4801 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4861 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4921 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4981 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5041 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5101 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5161 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5221 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5281 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5341 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5401 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5461 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5521 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5581 agcgaggaag cggaaga
//
