LOCUS       40924_40067        6979 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Recombinase expression vector pSH47, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6979)
  AUTHORS   Guldener U, Heck S, Fielder T, Beinhauer J, Hegemann JH.
  TITLE     A new efficient gene disruption cassette for repeated use in budding
            yeast
  JOURNAL   Nucleic Acids Res. 24 (13), 2519-2524 (1996)
  PUBMED    8692690
REFERENCE   2  (bases 1 to 6979)
  AUTHORS   Gueldener U, Heinisch J, Koehler GJ, Voss D, Hegemann JH.
  TITLE     A second set of loxP marker cassettes for Cre-mediated multiple gene
            knockouts in budding yeast
  JOURNAL   Nucleic Acids Res. 30 (6), E23 (2002)
  PUBMED    11884642
REFERENCE   3  (bases 1 to 6979)
  AUTHORS   Gueldener U, Hegemann JH, Heck S, Fiedler T, Beinhauer JD.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   4  (bases 1 to 6979)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 6979)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "1996"; volume: "24"; issue: "13"; pages: 
            "2519-2524"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        258..478
                     /label=URA3 promoter
     CDS             479..1279
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1413..1868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2013..2029
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2039..2057
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2076..2323)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2645..3673)
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(3771..3787)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3802..4243)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(4267..4285)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4306..4322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4330..4346)
                     /label=lac operator
                     /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(4354..4384)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4399..4420)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4708..5296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5470..6327)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6328..6432)
                     /label=AmpR promoter
     misc_feature    6469..6972
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
       61 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
      121 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
      181 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
      241 gcaccatacc acagcttttc aattcaattc atcatttttt ttttattctt ttttttgatt
      301 tcggtttctt tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
      361 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
      421 cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat
      481 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
      541 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
      601 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
      661 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
      721 cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt
      781 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
      841 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
      901 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
      961 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     1021 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     1081 acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga
     1141 tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     1201 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     1261 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     1321 acaaattaga gcttcaattt aattatatca gttattaccc tatgcggtgt gaaataccgc
     1381 acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa
     1441 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     1501 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     1561 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1621 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     1681 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1741 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     1801 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     1861 gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     1921 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     1981 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     2041 atacgactca ctatagggcg aattgggtac cggccgcaaa ttaaagcctt cgagcgtccc
     2101 aaaaccttct caagcaaggt tttcagtata atgttacatg cgtacacgcg tctgtacaga
     2161 aaaaaaagaa aaatttgaaa tataaataac gttcttaata ctaacataac tataaaaaaa
     2221 taaataggga cctagacttc aggttgtcta actccttcct tttcggttag agcggatgtg
     2281 gggggagggc gtgaatgtaa gcgtgacata actaattaca tgactcgagg tcgacaattc
     2341 caaccttacc caagagttcg ccaaactcag acatcacttt angcaaaacc gcgccgtgct
     2401 tcttcctcgg tggcattcat cacgaaatgt tcagcactac gcatactttt gacaggaaac
     2461 gcaacggata ttgagtcaat atcaggcatt ctatcgctca gctttacagt gacaatgacg
     2521 gctggcgact gaatattagt gcttacagac agcactacat attttccgtc gatgttgaaa
     2581 tcctttctca tatgtcacca taaatatcaa ataattatag caatcattta cgcgttaatg
     2641 gctaatcgcc atcttccagc aggcgcacca ttgcccctgt ttcactatcc agggtacgga
     2701 tatagttcat gacaatattt acattggtcc agccaccagc ttgcatgatc tccggtattg
     2761 aaactccagc gcgggccata tctcgcgcgg ctccgacacg ggcactgtgt ccagaccagg
     2821 ccaggtatct ctgaccagag tcatccttag cgccgtaaat caatcgatga gttgcttcaa
     2881 aaatcccttc cagggcgcga gttgatagct ggctggtggc agatggcgcg gcaacaccat
     2941 tttttctgac ccggcaaaac aggtagttat tcggatcatc agctacacca gagacggaaa
     3001 tccatcgctc gaccagttta gttaccccca ggctaagtgc cttctctaca cctgcggtgc
     3061 taaccagcgt tttcgttctg ccaatatgga ttaacattct cccaccgtca gtacgtgaga
     3121 tatctttaac cctgatcctg gcaatttcgg ctatacgtaa cagggtgtta taagcaatcc
     3181 ccagaaatgc cagattacgt atatcctggc agcgatcgct attttccatg agtgaacgaa
     3241 cctggtcgaa atcagtgcgt tcgaacgcta gagcctgttt tgcacgttca ccggcatcaa
     3301 cgttttcttt tcggatccgc cgcataacca gtgaaacagc attgctgtca cttggtcgtg
     3361 gcagcccgga ccgacgatga agcatgttta gctggcccaa atgttgctgg atagttttta
     3421 ctgccagacc gcgcgcctga agatatagaa gataatcgcg aacatcttca ggttctgcgg
     3481 gaaaccattt ccggttattc aacttgcacc atgccgccca cgaccggcaa acggacagaa
     3541 gcattttcca ggtatgctca gaaaacgcct ggcgatccct gaacatgtcc atcaggttct
     3601 tgcgaacctc atcactcgtt gcatcgaccg gtaatgcagg caaattttgg tgtacggtca
     3661 gtaaattgga catttaacac tcagataatg gttttaagta aagtgtacag gatcggctct
     3721 gcccctcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg gatccactag
     3781 ttctagaatc cggggttttt tctccttgac gttaaagtat agaggtatat taacaatttt
     3841 ttgttgatac ttttattaca tttgaataag aagtaataca aaccgaaaat gttgaaagta
     3901 ttagttaaag tggttatgca gtttttgcat ttatatatct gttaatagat caaaaatcat
     3961 cgcttcgctg attaattacc ccagaaataa ggctaaaaaa ctaatcgcat tatcatccta
     4021 tggttgttaa tttgattcgt tcatttgaag gtttgtgggg ccaggttact gccaattttt
     4081 cctcttcata accataaaag ctagtattgt agaatcttta ttgttcggag cagtgcggcg
     4141 cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac gaggacgcac ggaggagagt
     4201 cttccttcgg agggctgtca cccgctcggc ggcttctaat ccgtactaga gctccagctt
     4261 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
     4321 tgtgtgaaat tgttatccgc tcacaattcc acacaacata ggagccggaa gcataaagtg
     4381 taaagcctgg ggtgcctaat gagtgaggta actcacatta attgcgttgc gctcactgcc
     4441 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4501 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4561 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4621 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5461 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5521 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5581 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5641 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5701 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5761 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5821 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5881 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5941 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6001 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6061 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6121 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6181 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6241 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6301 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6361 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6421 aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat cacgtgctat
     6481 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     6541 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     6601 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     6661 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt
     6721 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     6781 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     6841 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     6901 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     6961 aggcaagcga tccgtccta
//
