LOCUS       40924_40097        7346 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Recombinase expression vector pSH67, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7346)
  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 7346)
  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 7346)
  AUTHORS   Hegemann JH, Heick SB.
  TITLE     Delete and Repeat: A Comprehensive Toolkit for Sequential Gene 
            Knockout in the Budding Yeast Saccharomyces cerevisiae
  JOURNAL   Methods Mol. Biol. 765, 189-206 (2011)
  PUBMED    21815094
REFERENCE   4  (bases 1 to 7346)
  AUTHORS   Hegemann JH, Heick SB.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung 
            der Mikroorganismen, Heinrich-Heine-Universitaet, 
            Universitatsstrasse 1, Duesseldorf, NRW 40225, Germany
REFERENCE   5  (bases 1 to 7346)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 7346)
  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: "Methods 
            Mol. Biol. 765, 189-206 (2011)"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: "Submitted 
            (15-OCT-2010) Lehrstuhl fur Funktionelle Genomforschung der 
            Mikroorganismen, Heinrich-Heine-Universitaet, Universitatsstrasse 1,
            Duesseldorf, NRW 40225, Germany"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            241..1597
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     rep_origin      complement(1718..2173)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2318..2334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2344..2362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2381..2628)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2950..3978)
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(4076..4092)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4107..4548)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(4572..4590)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4611..4627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4635..4651)
                     /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(4659..4689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4704..4725)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5013..5601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5775..6632)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6633..6737)
                     /label=AmpR promoter
     misc_feature    6774..7277
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatagg tctagagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc
      241 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga
      301 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata
      361 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag
      421 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga
      481 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc
      541 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga
      601 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
      661 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg
      721 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
      781 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
      841 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc
      901 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
      961 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1021 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1081 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat
     1141 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1201 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1261 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1321 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1381 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     1441 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     1501 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     1561 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     1621 ccatccagtg tcgaaaacga gctctcgaga acccttaatc ggatctatgc ggtgtgaaat
     1681 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaaacgt taatattttg
     1741 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     1801 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     1861 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     1921 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     1981 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     2041 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     2101 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     2161 ctacagggcg cgtcgcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     2221 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     2281 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
     2341 gcgtaatacg actcactata gggcgaattg ggtaccggcc gcaaattaaa gccttcgagc
     2401 gtcccaaaac cttctcaagc aaggttttca gtataatgtt acatgcgtac acgcgtctgt
     2461 acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac ataactataa
     2521 aaaaataaat agggacctag acttcaggtt gtctaactcc ttccttttcg gttagagcgg
     2581 atgtgggggg agggcgtgaa tgtaagcgtg acataactaa ttacatgact cgaggtcgac
     2641 aattccaacc ttacccaaga gttcgccaaa ctcagacatc actttangca aaaccgcgcc
     2701 gtgcttcttc ctcggtggca ttcatcacga aatgttcagc actacgcata cttttgacag
     2761 gaaacgcaac ggatattgag tcaatatcag gcattctatc gctcagcttt acagtgacaa
     2821 tgacggctgg cgactgaata ttagtgctta cagacagcac tacatatttt ccgtcgatgt
     2881 tgaaatcctt tctcatatgt caccataaat atcaaataat tatagcaatc atttacgcgt
     2941 taatggctaa tcgccatctt ccagcaggcg caccattgcc cctgtttcac tatccagggt
     3001 acggatatag ttcatgacaa tatttacatt ggtccagcca ccagcttgca tgatctccgg
     3061 tattgaaact ccagcgcggg ccatatctcg cgcggctccg acacgggcac tgtgtccaga
     3121 ccaggccagg tatctctgac cagagtcatc cttagcgccg taaatcaatc gatgagttgc
     3181 ttcaaaaatc ccttccaggg cgcgagttga tagctggctg gtggcagatg gcgcggcaac
     3241 accatttttt ctgacccggc aaaacaggta gttattcgga tcatcagcta caccagagac
     3301 ggaaatccat cgctcgacca gtttagttac ccccaggcta agtgccttct ctacacctgc
     3361 ggtgctaacc agcgttttcg ttctgccaat atggattaac attctcccac cgtcagtacg
     3421 tgagatatct ttaaccctga tcctggcaat ttcggctata cgtaacaggg tgttataagc
     3481 aatccccaga aatgccagat tacgtatatc ctggcagcga tcgctatttt ccatgagtga
     3541 acgaacctgg tcgaaatcag tgcgttcgaa cgctagagcc tgttttgcac gttcaccggc
     3601 atcaacgttt tcttttcgga tccgccgcat aaccagtgaa acagcattgc tgtcacttgg
     3661 tcgtggcagc ccggaccgac gatgaagcat gtttagctgg cccaaatgtt gctggatagt
     3721 ttttactgcc agaccgcgcg cctgaagata tagaagataa tcgcgaacat cttcaggttc
     3781 tgcgggaaac catttccggt tattcaactt gcaccatgcc gcccacgacc ggcaaacgga
     3841 cagaagcatt ttccaggtat gctcagaaaa cgcctggcga tccctgaaca tgtccatcag
     3901 gttcttgcga acctcatcac tcgttgcatc gaccggtaat gcaggcaaat tttggtgtac
     3961 ggtcagtaaa ttggacattt aacactcaga taatggtttt aagtaaagtg tacaggatcg
     4021 gctctgcccc tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc
     4081 actagttcta gaatccgggg ttttttctcc ttgacgttaa agtatagagg tatattaaca
     4141 attttttgtt gatactttta ttacatttga ataagaagta atacaaaccg aaaatgttga
     4201 aagtattagt taaagtggtt atgcagtttt tgcatttata tatctgttaa tagatcaaaa
     4261 atcatcgctt cgctgattaa ttaccccaga aataaggcta aaaaactaat cgcattatca
     4321 tcctatggtt gttaatttga ttcgttcatt tgaaggtttg tggggccagg ttactgccaa
     4381 tttttcctct tcataaccat aaaagctagt attgtagaat ctttattgtt cggagcagtg
     4441 cggcgcgagg cacatctgcg tttcaggaac gcgaccggtg aagacgagga cgcacggagg
     4501 agagtcttcc ttcggagggc tgtcacccgc tcggcggctt ctaatccgta ctagagctcc
     4561 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     4621 tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc cggaagcata
     4681 aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc gttgcgctca
     4741 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4801 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt
     6781 gctataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
     6841 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
     6901 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
     6961 gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt
     7021 tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat
     7081 atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt
     7141 ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata
     7201 aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa
     7261 gttacaggca agcgatccgt cctaagaaac cattattatc atgacattaa cctataaaaa
     7321 taggcgtatc acgaggccct ttcgtc
//
