LOCUS       sequence-247535-        5598 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 5598)
  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 5598)
  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..5598
                     /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..554
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIYRWAILMKN
                     ILNLLSYY"
     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             1262..1270
                     /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(3366..3581)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3606..3624)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3692..3796
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3797..3865,3866..4657)
                     /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(4015..4034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4828..5416
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5317..5336
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5570..5587
                     /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 tatcgctggg ccattctcat gaagaatatc ttgaatttat
      541 tgtcatatta ctagtaggca tttgtttatt accagcttac gatataaagt cgttcatctt
      601 ctttatggcg agaaactgta attgcgaaaa ttgtctaggc aatttttttc agtagttagt
      661 ctgagatttt tccttgtgaa ccaaccttcc tgtcgttatt aatactagaa gataagacga
      721 aacagtttag tttgtaggta cgtaaaaact tgttcaaagt aggcttcagt aatcttgttc
      781 cgaaaaagtt tctcacatat caaacaaatt agaggtttcg gaagtaagcc acaacaaaaa
      841 taatgaatat tttttaaacg cttttaaaac agataattat caaaaaatag aattaaagct
      901 ttatttgtat taattttgaa agtattaaat aaaattatat actgtcttat aatatatatt
      961 attatatcat gtgattaaat tacacagctt ctaaagctct ttaatggatt gcggttttta
     1021 catctactct actttagatc atactttgtc tttgattaaa gtatattaag tggccatctg
     1081 tgaccagtat cttttgaaac aatttgtcat aatatccata ttccatcata ttaaacattt
     1141 acgttttgtt ctactttaac caatttgaaa gacattattt aaatcaaaaa tattttctcc
     1201 gagttagcta tgaaaaagat tgaatacctc atgtataaag atatcctctc cttccaagaa
     1261 tacctcctta agtttcattt ctaccgtcag gactcgtgtt ttcgccagga gcgataaggg
     1321 gacgtgtgat ttctgcatca tcgttattag gtgacactaa gttagtgccc gatcgatgct
     1381 agcaagaatc gggtcattgt agcgtatgcg cctgtgaaca ttctcttcaa caagtttgat
     1441 tccattgcgg tgaaatggta aaagtcaacc ccctgcgatg tatattttcc tgtacaatca
     1501 atcaaaaagc caaatgattt agcattatct ttacatcttg ttattttaca gattttatgt
     1561 ttagatcttt tatgcttgct tttcaaaagg cttgcaggca agtgcacaaa caatacttaa
     1621 ataaatacta ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt
     1681 tagagtcttt tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat
     1741 ttaatctaag cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta
     1801 agctctcggg gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt
     1861 cacctgtccc acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag
     1921 ctgcactgag tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac
     1981 cgaggaactc ttggtattct tgccacgact catctccatg cagttggacg atcgatgata
     2041 agctgtcaaa catgagaatt gggtaataac tgatataatt aaattgaagc tctaatttgt
     2101 gagtttagta tacatgcatt tacttataat acagttttat ggacggtggt atgttttagt
     2161 ttagcttcaa ttctaattga ttgattaatc agttgattgg tttcaatatg acaaatgggt
     2221 agggtgggaa aacttcattt tcaattcaga tcaaactttt ttgttgtcga cataatattt
     2281 ctcgtttggg atgttactgt cacattaata atacacacac atcagcttat aattttgaaa
     2341 gtaatttatc agatatgttg tgacgatcag tggaaatggc taacttcaat gtatctgttc
     2401 ttcccctttt tcaaagttca cgttttttga ttgattgatt gatctgtcgg cagtggtttc
     2461 aaaaccattc ggtgagtaat cctatcaatc aatgttacga caaaaggctc aatattcaaa
     2521 attgcaatgt tttatgtttt cctacgtgta cttgtgcaag gcaattgatt caacattgct
     2581 tttggtgttt gacgagtttc tagtttggac ttgtgttgtt atctgggcta tacagatttc
     2641 ccggctcact atgaattttt tttttcgacg ctcagtgcac acaactataa acaacacaaa
     2701 cacaaacaca gcaagaaaaa aaaaaaacga acattgaatt gaaaccaagc caactgaaaa
     2761 attccttatt taaatgactg tcatactaac ccatttttat agaagaagtt gctgctttag
     2821 ttatcgataa cggttctcat atgaaaattt cggtgatccc tgagcaggtg gcggaaacat
     2881 tggatgctga gaaccatttc attgttcgtg aagtgttcga tgtgcaccta tccgaccaag
     2941 gctttgaact atctaccaga agtgtgagcc cctaccggaa ggattacatc tcggatgatg
     3001 actctgatga agactctgct tgctatggcg cattcatcga ccaagagctt gtcgggaaga
     3061 ttgaactcaa ctcaacatgg aacgatctag cctctatcga acacattgtt gtgtcgcaca
     3121 cgcaccgagg caaaggagtc gcgcacagtc tcatcgaatt tgcgaaaaag tgggcactaa
     3181 gcagacagct ccttggcata cgattagaga cacaaacgaa caatgtacct gcctgcaatt
     3241 tgtacgcaaa atgtggcttt actctcggcg gcattgacct cttcacgtat aaaactagac
     3301 ctcaagtctc gaacgaaaca gcgatgtact ggtactggtt ctcgggagca caggatgacg
     3361 cctaagattt atcttcgttt cctgcatgtt tttgttctgt gcagttgggt taagaatact
     3421 gggcaatttc atgtttcttc aacactacat atgcgtatat ataccaatct aagtctgtgc
     3481 tccttccttc gttcttcctt ctgttcggag attaccgaat caaaaaaatt tcaaggaaac
     3541 cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga aaagctaatt cttgaagacg
     3601 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3661 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3721 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3781 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3841 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3901 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3961 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4021 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4081 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     4141 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4201 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4261 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4321 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     4381 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4441 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4501 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4561 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4621 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4681 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4741 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4801 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4861 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4921 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4981 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5041 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5101 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5161 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5221 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5281 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5341 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5401 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5461 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5521 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5581 gagcgaggaa gcggaaga
//
