LOCUS       sequence-252444-        7828 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cre recombinase in Z. rouxii yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7828)
  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 7828)
  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: pGRCRE
FEATURES             Location/Qualifiers
     source          1..7828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        666..737
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(738..806,807..1598)
                     /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(956..975)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1769..2357
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2258..2277
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2511..2528
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2645..2666
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2681..2686,2687..2704,2705..2711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2719..2735
                     /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     2724..2746
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2743..2759
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2743..2759
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2778..2798
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2780..2798
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2834..3275
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2834..2951
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3246..3269
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     3290..3306
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3290..3306
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3404..4435
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(4816..4835)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4817..4835)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4845..4862)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4845..4861)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4854..4876)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5002..5457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5089..5108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5299..5320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7659..7678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7778..7800
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt
      121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag
      181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt
      241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat
      301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt
      361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa
      421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac
      481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt
      541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca
      601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat
      661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2641 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2701 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcgaat
     2821 tcgagctcta gtacggatta gaagccgccg agcgggtgac agccctccga aggaagactc
     2881 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
     2941 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
     3001 attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa caaccatagg
     3061 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
     3121 tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt aactaatact
     3181 ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat caacaaaaaa
     3241 ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatt ctagaactag
     3301 tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg aggggcagag
     3361 ccgatcctgt acactttact taaaaccatt atctgagtgt taaatgtcca atttactgac
     3421 cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa
     3481 cctgatggac atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct
     3541 gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc
     3601 agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa
     3661 aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg
     3721 accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga
     3781 tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg
     3841 ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat
     3901 tgcttataac accctgttac gtatagccga aattgccagg atcagggtta aagatatctc
     3961 acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac
     4021 cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc
     4081 cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg
     4141 tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga
     4201 agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg
     4261 gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat
     4321 accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg
     4381 taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg attagccata
     4441 tatatataac gtacacatag aaatcacaca aacagagtat ttattcttaa ctacatgaac
     4501 taccatcaga ccgtctgggc cactatataa tgtgccattc ataaacgtga tcactttacg
     4561 tagcaggcaa ccccaggtga aaatttttca gcgagctgcc agattgtcag gtgaaaaact
     4621 gaaaaaaact tctgggcgat gagcttgtgg cgggaaatta agtatataaa gcagttagtt
     4681 tcttctgctt cttgtggttc tgggtattct tgtaatattt ggaagaatag aaaatcaaga
     4741 ctactgcctt tcttttcata ttatagagga agaaatacgc acgaacacga tatagaggta
     4801 aaggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta
     4861 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     4921 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     4981 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     5041 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     5101 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     5161 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     5221 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     5281 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     5341 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5401 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     5461 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag ggtaataact
     5521 gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt acttataata
     5581 cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccccttt gttgttgttg
     5641 tgggtgtgtg ctatttatat tatatagttt actcataata ccataaaata ttcggtttga
     5701 ttaggttatt ttaataagct aatttgtttc taatcgtgta atttatgctg tgtatattaa
     5761 gtagtgtgtg cactgcccaa aaatgtttgt tgtttatagt cggttaaaga gaaaaaagaa
     5821 aaaaagatcc atacacacac gttaattagt tgttcaacgt aatacactca tattttgttc
     5881 ttatttgctt tcggtcgctg ttctcaccaa gatttattgc caacgaaaca attttttttt
     5941 atatattttc agatttttct ttttttcctt tcctttcctt ttctaatttt cactcctggt
     6001 tttctttctt tcttagaaac attatctcga tattaatatt aaaaaaatat aatcattcaa
     6061 aatggacggt ggtatgtttt agtttagctt caattctaat tgattgatta atcagttgat
     6121 tggtttcaat atgacaaatg ggtagggtgg gaaaacttca ttttcaattc agatcaaact
     6181 tttttgttgt cgacataata tttctcgttt gggatgttac tgtcacatta ataatacaca
     6241 cacatcagct tataattttg aaagtaattt atcagatatg ttgtgacgat cagtggaaat
     6301 ggctaacttc aatgtatctg ttcttcccct ttttcaaagt tcacgttttt tgattgattg
     6361 attgatctgt cggcagtggt ttcaaaacca ttcggtgagt aatcctatca atcaatgtta
     6421 cgacaaaagg ctcaatattc aaaattgcaa tgttttatgt tttcctacgt gtacttgtgc
     6481 aaggcaattg attcaacatt gcttttggtg tttgacgagt ttctagtttg gacttgtgtt
     6541 gttatctggg ctatacagat ttcccggctc actatgaatt ttttttttcg acgctcagtg
     6601 cacacaacta taaacaacac aaacacaaac acagcaagaa aaaaaaaaaa cgaacattga
     6661 attgaaacca agccaactga aaaattcctt atttaaatga ctgtcatact aacccatttt
     6721 tatagaagaa gttgctgctt tagttatcga taacggttct catatgaaaa tttcggtgat
     6781 ccctgagcag gtggcggaaa cattggatgc tgagaaccat ttcattgttc gtgaagtgtt
     6841 cgatgtgcac ctatccgacc aaggctttga actatctacc agaagtgtga gcccctaccg
     6901 gaaggattac atctcggatg atgactctga tgaagactct gcttgctatg gcgcattcat
     6961 cgaccaagag cttgtcggga agattgaact caactcaaca tggaacgatc tagcctctat
     7021 cgaacacatt gttgtgtcgc acacgcaccg aggcaaagga gtcgcgcaca gtctcatcga
     7081 atttgcgaaa aagtgggcac taagcagaca gctccttggc atacgattag agacacaaac
     7141 gaacaatgta cctgcctgca atttgtacgc aaaatgtggc tttactctcg gcggcattga
     7201 cctcttcacg tataaaacta gacctcaagt ctcgaacgaa acagcgatgt actggtactg
     7261 gttctcggga gcacaggatg acgcctaaca tatgtgaagt gtgaaggggg agattttcac
     7321 tttattagat ttgtatatat gtataataaa taaataaata agttaaataa ataattagat
     7381 aagggtggta attattacta tttacaatca aaggtggtcc tgcaggcttc gtttcctgca
     7441 ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     7501 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     7561 ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa
     7621 aaatgatgaa ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca
     7681 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     7741 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     7801 ttttcaccgt catcaccgaa acgcgcga
//
