LOCUS       sequence-12721-e        7226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of yeast codon-optimized CreA recombinase from the GAL1 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7226)
  AUTHORS   Fang F, Salmon K, Shen MW, Aeling KA, Ito E, Irwin B, Tran UP, 
            Hatfield GW, Da Silva NA, Sandmeyer S
  TITLE     A vector set for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2011 Feb;28(2):123-36. doi: 10.1002/yea.1824. Epub 2010 Oct 
            8.
  PUBMED    20936606
REFERENCE   2  (bases 1 to 7226)
  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.1824"; 
            journalName: "Yeast"; date: "2011-02"; volume: "28"; issue: "2"; 
            pages: "123-36"
COMMENT     Sequence Label: pBF3060
FEATURES             Location/Qualifiers
     source          1..7226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..443
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2..119
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     414..437
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      1627..1874
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1643..1661)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1944..1961)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2115..2703)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2195..2214)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2874..3665,3666..3734))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(3830..4633)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4634..4854)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      5805..6685
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(6754..7209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7103..7122
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
       61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
      121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
      181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
      241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
      301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
      361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
      421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gccatggaaa
      481 aaaatgtcta atttgttgac tgttcatcaa aatttgccag ctttgccagt tgatgcaacg
      541 tctgatgagg taagaaagaa tttgatggat atgtttagag atagacaagc attctctgaa
      601 catacttgga aaatgttgtt gtctgtttgt agatcttggg ctgcttggtg taaattaaat
      661 aatagaaaat ggtttccagc tgaaccagaa gatgttagag attatttgtt gtatttgcaa
      721 gctagaggtt tggctgttaa aactatacaa caacatttgg gtcaattaaa tatgttgcat
      781 aggagatctg gtttgcctag accatctgat tctaatgctg tatctttggt tatgagaagg
      841 attagaaaag aaaatgttga tgcaggtgaa agagctaaac aagcattggc gtttgaaaga
      901 actgattttg atcaagttag atctttgatg gaaaattctg atagatgtca agatataaga
      961 aacttggctt tcttgggaat tgcttataat acattgttga gaattgctga aattgctaga
     1021 attagagtta aagacatttc tagaactgat ggtggtagaa tgttgattca tattggtaga
     1081 actaaaacat tggtttcaac tgctggcgtt gagaaagcat tgtctttggg tgttactaaa
     1141 ttggttgaaa gatggatttc tgtatctggt gtggctgatg acccaaataa ttatttattt
     1201 tgtagagtta gaaagaatgg tgttgcggct ccatctgcta catctcagct ctctacccgt
     1261 gccttggaag gaatttttga agcaactcat agattgattt atggtgctaa agatgattct
     1321 ggtcaaagat atttagcatg gtctggtcat tctgctagag ttggtgccgc tagagatatg
     1381 gctagagctg gggtatctat tcctgaaatt atgcaagctg gtggttggac taatgttaac
     1441 atcgtcatga actatattag aaatttggat tctgaaactg gtgcaatggt tagactcttg
     1501 gaagatggtg atcccgggaa gcttggtacc gagctcggat ccactagtaa cggccgccag
     1561 tgtgctggaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg
     1621 gccgcatcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     1681 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     1741 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     1801 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     1861 aggctttaat ttgcggccct gcattaatga atcggccaac gcgcggggag aggcggtttg
     1921 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1981 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2041 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagc ccaggaaccg taaaaaggcc
     2101 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2161 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2221 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2281 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2341 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2401 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2461 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2521 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2581 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2641 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2701 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2761 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2821 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     2881 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     2941 tgactccccg tcgtgtagat aactacgata cgggagcgct taccatctgg ccccagtgct
     3001 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3061 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat tcagtctatt
     3121 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3181 ggcattgcta caggcatcgt ggtgtcactc tcgtcgtttg gtatggcttc attcagctcc
     3241 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3301 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     3361 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     3421 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3481 ccggcgtcaa tacgggataa tagtgtatca catagcagaa ctttaaaagt gctcatcatt
     3541 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     3601 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3661 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3721 tgttgaatac tcatactctt cctttttcaa tgggtaataa ctgatataat taaattgaag
     3781 ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct
     3841 ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct
     3901 tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag
     3961 agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac
     4021 ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt
     4081 gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag
     4141 tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc
     4201 taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca
     4261 ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt
     4321 actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat
     4381 tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca
     4441 agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca
     4501 gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca
     4561 tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg
     4621 tagctttcga catgatttat cttcgtttcc tgcaggtttt tgttctgtgc agttgggtta
     4681 agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa
     4741 gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc
     4801 aaagaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat tgaaaagcta
     4861 gcttatcgat gataagctgt caaagatgag aattaattcc acggactata gactatacta
     4921 gatactccgt ctactgtacg atacacttcc gctcaggtcc ttgtccttta acgaggcctt
     4981 accactcttt tgttactcta ttgatccagc tcagcaaagg cagtgtgatc taagattcta
     5041 tcttcgcgat gtagtaaaac tagctagacc gagaaagaga ctagaaatgc aaaaggcact
     5101 tctacaatgg ctgccatcat tattatccga tgtgacgctg cagcttctca atgatattcg
     5161 aatacgcttt gaggagatac agcctaatat ccgacaaact gttttacaga tttacgatcg
     5221 tagataagct gtcaaagatg agaattaatt ccacggacta tagactatac tagatactcc
     5281 gtctactgta cgatacactt ccgctcaggt ccttgtcctt taacgaggcc ttaccactct
     5341 tttgttactc tattgatcca gctcagcaaa ggcagtgtga tctaagattc tatcttcgcg
     5401 atgtagtaaa actagctaga ccgagaaaga gactagaaat gcaaaaggca cttctacaat
     5461 ggctgccatc attattatcc gatgtgacgc tgcagcttct caatgatatt cgaatacgct
     5521 ttgaggagat acagcctaat atccgacaaa ctgttttaca gatttacgat cgtacttgtt
     5581 acccatcatt gaattttgaa catccgaacc tgggagtttt ccctgaaaca gatagtatat
     5641 ttgaacctgt ataataatat atagtctagc gctttacgga agacaatgta tgtatttcgg
     5701 ttcctggaga aactattgca tctattgcat aggtaatctt gcacgtcgca tccccggttc
     5761 attttctgcg tttccatctt gcacttcaat agcatatctt tgttaacgaa gcatctgtgc
     5821 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     5881 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct
     5941 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgacgaga gcgctaattt ttcaaacaaa
     6001 gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca
     6061 aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa
     6121 caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata
     6181 actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc
     6241 ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg
     6301 tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat
     6361 actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg
     6421 gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt
     6481 ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag taatactaga
     6541 gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga
     6601 tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa
     6661 tgtttgtgga agcggtattc gcaatgggaa gctccacccc ggttgataat cagaaaagcc
     6721 ccaaaaacag gaagattgta taagcaaata tttaaattgt aaacgttaat attttgttaa
     6781 aattcgcgtt aaatttttgt taaatcagct cattttttaa cgaatagccc gaaatcggca
     6841 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttcca
     6901 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa agggtctatc
     6961 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     7021 gtaaagcagt aaatcggaag ggtaaacgga tgcccccatt tagagcttga cggggaaagc
     7081 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggggct agggcggtgg
     7141 gaagtgtagg ggtcacgctg ggcgtaacca ccacacccgc cgcgcttaat ggggcgctac
     7201 agggcgcgtg gggatgatcc actagt
//
