LOCUS       40924_40082        6869 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Recombinase expression vector pSH63, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6869)
  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   2  (bases 1 to 6869)
  AUTHORS   Gueldener U, Heinisch JH, Voss D, Koehler G, Hegemann JH.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   3  (bases 1 to 6869)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6869)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2002"; volume: "30"; issue: "6"; pages: "E23"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        187..467
                     /label=TRP1 promoter
     CDS             468..1139
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1904..2151)
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(2473..3501)
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(3599..3615)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3630..4071)
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     promoter        complement(4095..4113)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4134..4150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4158..4174)
                     /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(4182..4212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4227..4248)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4536..5124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5298..6155)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6156..6260)
                     /label=AmpR promoter
     misc_feature    6297..6800
                     /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 tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg gccgcaaatt aaagccttcg
     1921 agcgtcccaa aaccttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc
     1981 tgtacagaaa aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta
     2041 taaaaaaata aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag
     2101 cggatgtggg gggagggcgt gaatgtaagc gtgacataac taattacatg actcgaggtc
     2161 gacaattcca accttaccca agagttcgcc aaactcagac atcactttan gcaaaaccgc
     2221 gccgtgcttc ttcctcggtg gcattcatca cgaaatgttc agcactacgc atacttttga
     2281 caggaaacgc aacggatatt gagtcaatat caggcattct atcgctcagc tttacagtga
     2341 caatgacggc tggcgactga atattagtgc ttacagacag cactacatat tttccgtcga
     2401 tgttgaaatc ctttctcata tgtcaccata aatatcaaat aattatagca atcatttacg
     2461 cgttaatggc taatcgccat cttccagcag gcgcaccatt gcccctgttt cactatccag
     2521 ggtacggata tagttcatga caatatttac attggtccag ccaccagctt gcatgatctc
     2581 cggtattgaa actccagcgc gggccatatc tcgcgcggct ccgacacggg cactgtgtcc
     2641 agaccaggcc aggtatctct gaccagagtc atccttagcg ccgtaaatca atcgatgagt
     2701 tgcttcaaaa atcccttcca gggcgcgagt tgatagctgg ctggtggcag atggcgcggc
     2761 aacaccattt tttctgaccc ggcaaaacag gtagttattc ggatcatcag ctacaccaga
     2821 gacggaaatc catcgctcga ccagtttagt tacccccagg ctaagtgcct tctctacacc
     2881 tgcggtgcta accagcgttt tcgttctgcc aatatggatt aacattctcc caccgtcagt
     2941 acgtgagata tctttaaccc tgatcctggc aatttcggct atacgtaaca gggtgttata
     3001 agcaatcccc agaaatgcca gattacgtat atcctggcag cgatcgctat tttccatgag
     3061 tgaacgaacc tggtcgaaat cagtgcgttc gaacgctaga gcctgttttg cacgttcacc
     3121 ggcatcaacg ttttcttttc ggatccgccg cataaccagt gaaacagcat tgctgtcact
     3181 tggtcgtggc agcccggacc gacgatgaag catgtttagc tggcccaaat gttgctggat
     3241 agtttttact gccagaccgc gcgcctgaag atatagaaga taatcgcgaa catcttcagg
     3301 ttctgcggga aaccatttcc ggttattcaa cttgcaccat gccgcccacg accggcaaac
     3361 ggacagaagc attttccagg tatgctcaga aaacgcctgg cgatccctga acatgtccat
     3421 caggttcttg cgaacctcat cactcgttgc atcgaccggt aatgcaggca aattttggtg
     3481 tacggtcagt aaattggaca tttaacactc agataatggt tttaagtaaa gtgtacagga
     3541 tcggctctgc ccctcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga
     3601 tccactagtt ctagaatccg gggttttttc tccttgacgt taaagtatag aggtatatta
     3661 acaatttttt gttgatactt ttattacatt tgaataagaa gtaatacaaa ccgaaaatgt
     3721 tgaaagtatt agttaaagtg gttatgcagt ttttgcattt atatatctgt taatagatca
     3781 aaaatcatcg cttcgctgat taattacccc agaaataagg ctaaaaaact aatcgcatta
     3841 tcatcctatg gttgttaatt tgattcgttc atttgaaggt ttgtggggcc aggttactgc
     3901 caatttttcc tcttcataac cataaaagct agtattgtag aatctttatt gttcggagca
     3961 gtgcggcgcg aggcacatct gcgtttcagg aacgcgaccg gtgaagacga ggacgcacgg
     4021 aggagagtct tccttcggag ggctgtcacc cgctcggcgg cttctaatcc gtactagagc
     4081 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
     4141 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc
     4201 ataaagtgta aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc
     4261 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4321 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5221 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5401 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5581 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5881 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6241 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc cttttcatca
     6301 cgtgctataa aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag
     6361 aaagtaaaaa aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc
     6421 tagggggatc gccaacaaat actacctttt atcttgctct tcctgctctc aggtattaat
     6481 gccgaattgt ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca
     6541 ttttacttat cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata
     6601 tatatgtaaa gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc
     6661 attccgtaac tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa
     6721 ataaataaac acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg
     6781 taagttacag gcaagcgatc cgtcctaaga aaccattatt atcatgacat taacctataa
     6841 aaataggcgt atcacgaggc cctttcgtc
//
