LOCUS       V003681                 7357 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003681
VERSION     V003681
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7357)
  AUTHORS   Redder P, Linder P.
  TITLE     New Range of Vectors with a Stringent 5-Fluoroorotic Acid-Based
            Counterselection System for Generating Mutants by Allelic
            Replacement in Staphylococcus aureus
  JOURNAL   Appl. Environ. Microbiol. 78 (11), 3846-3854 (2012)
   PUBMED   22447609
REFERENCE   2  (bases 1 to 7357)
  AUTHORS   Redder P, Linder P.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAR-2012) Department of Microbiology and Molecular
            Medicine, University of Geneva, Faculty of Medicine, Rue
            Michel-Servet 1, Geneva 1203, Switzerland
REFERENCE   3  (bases 1 to 7357)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7357)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: BioEdit, CAP v. 7.0.9.0
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2012"; volume: "78"; issue: "11";
            pages: "3846-3854"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (12-MAR-2012) Department of Microbiology and Molecular Medicine,
            University of Geneva, Faculty of Medicine, Rue Michel-Servet 1,
            Geneva 1203, Switzerland"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    396..452
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(465..481)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(489..505)
                     /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(513..543)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(558..579)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(717..1364)
                     /gene="pyrE"
                     /label="Orotate phosphoribosyltransferase"
                     /note="Orotate phosphoribosyltransferase from Bacillus
                     subtilis (strain 168). Accession#: P25972"
     CDS             complement(1346..2062)
                     /gene="pyrF"
                     /label="Orotidine 5'-phosphate decarboxylase"
                     /note="Orotidine 5'-phosphate decarboxylase from Bacillus
                     subtilis (strain 168). Accession#: P25971"
     CDS             complement(2450..3826)
                     /gene="tet"
                     /label="Tetracycline resistance protein"
                     /note="Tetracycline resistance protein from Staphylococcus
                     aureus. Accession#: P02983"
     rep_origin      complement(4233..5397)
                     /direction=LEFT
                     /note="thermosensitive origin of replication; for
                     replication in Staphylococcus aureus; pE194ts"
     rep_origin      complement(5538..6126)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6300..7157)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7158..7262)
                     /label="AmpR promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct gtcgaatggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata gctgtttcct
      481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
      601 gctttccagt cgggaaccac tcgagccatg gattaataaa aaaacaggcc tcacagcctg
      661 ttttcaagta cagcaaggct ttgaagcctc tttacatcat ttgaatttat tttttatttt
      721 ttaaaccaat cttttgactc gggatttcgt ttccatgttt gcagcttttt tagatcatct
      781 gaatgaatat ttccgttttc aagcgcgacc tctgtgagcg tatcataatc ggttaatgag
      841 tagtatggca gttctgcctt tgcgaaggct tcctccgctt taggaagtcc gtacgtaaag
      901 attgagacga caccaagcac ttcacagccg gccgcttgta aagctgcaca ggcttcaagc
      961 acgctgcctc ctgtggaaat taagtcttca atgacgactg ttttttgccc ttcttgcaca
     1021 gctccctcaa tctgattgcc ttttccgtgc gccttcggct tgctcctcac ataacacatc
     1081 ggaagattca aatggtccgc cgcaagagca gcatgaggaa taccggcagt tgctgttccc
     1141 gcaatcattt cagcttcagg aaaatgctct ttaacaagtt tgctgatacc tgaagcaaca
     1201 tcgtttctga cctctgggaa agatagcgta aggcggttgt cacagtagat cggtgataaa
     1261 atgccgcttg cccatgtgaa cggctcgttc gggcgtaaaa atacagcttg gatgtctaat
     1321 agatgttttg cgatgatttg tttcaagatt tgattccctc ccattcaagt ctgacagcct
     1381 tataggcttt taccgggtct tccgcttttg taatcgagcg tcctactaca atcgctgatg
     1441 aacctttctc tcttgcaatg gcaggcgtcg ctacgcgaac ttggtcattc gcagcgtcct
     1501 ctgacattct gatccccgga gtgacagtca gaaatgaagg cgacaccgct tggtaaatgg
     1561 cttttgcttc atgaacagag cagaccactc catccagtcc gctttcttcc gcctgtttgc
     1621 tgtagtgcac aaccgtatca atcagagact tttcgatcag cagttcatct ttcatgattt
     1681 gctcagatgt gcttgtcagc tgggttaccg cgataagtga cggacgtttt tttccagccg
     1741 gcgtaccttc ttctaagcct tcgagagctg cctgcatcat ttttttgccc ccggcagcat
     1801 gaacattgac gaggtctact ccaagactgg caaggcgctt catcgctttg tttacagtag
     1861 tcgggatgtc atgaagtttt aagtctaaaa atagctcgca gtttctttct tttagttgtt
     1921 tcacgataga tggcccttct tgataaaaaa gctccatccc aacctttaca aataacggtt
     1981 cttgctgaaa aggcgctaag aacgcaagtg tttcttcagc tgacgcaaaa tcaagcgcga
     2041 tgatgggcag gttgtttttc atgattccag ctccttccga ttccatacat tatgccacac
     2101 cttgtagata aagtcaacaa ctttttgcaa aatttttcag gaattttagc agaggttgtt
     2161 ctggatgtag aacaaaacat ctttccgctc ttgtgctgtt aggatatctt tcttggaagc
     2221 taggtaggca agggctacct caaataaatc ttcttcaggg tgcgctattt ttaaggtgcc
     2281 tagtgaggtc ttgaccactt cacccataat ttcagtgccg aatagtctgg actgggctgt
     2341 gtagatctta cgtgtgctct gcgaggctta aacctgtttt tgctaacgca aaaaatgagt
     2401 ggcaaaatgc tagccactca tagttctaaa ccaaaatata atataactat tcaaactgct
     2461 tttcagaacg tttaaataca ataatcgtca aaagacaaca taaaataata aggatagcca
     2521 tggctacaag aatattacta tacactccag aagaataatt tataaattcc agaactagtt
     2581 tacgattaat caattgtagt gacaataaac ctcctacaat tgctatacct gttccctctg
     2641 ataaaaaact tgtgaaattt agcaaactca ttccagaagc aacttcttct tcagaaagac
     2701 tacttgatac tatttttgat ataactgttt tagtaaaaga taatccgccc ataacaaata
     2761 taaacataaa agtagtcaac cacatactaa actcaacaaa aaatgcaata gttaaaaaac
     2821 ttatagagat agacaatgat cctaaaataa aaacaaataa tgatcctttt ctatccacta
     2881 aaaaaccacc aaaataacca aaaacaataa cactcatggt tccaggaaaa ataacactat
     2941 tacctattgt cgctacattt acatgataaa tagttttcat catataaggc accattgata
     3001 taaaaccagc tactatagaa aatattagcc caccagaaaa caaaccaagc ataaacggaa
     3061 tgtttttccc tagtttagga ttaataaaag ggttagaaac tcttgaaata tgtttaataa
     3121 aaatcacaaa aaagattgtg aagagtatta aaaaagtcca attataattt gtcgtaaata
     3181 acataaaaca tataatactt atagacatta aaacaatacc tacgatatct aatgtatttt
     3241 ttgttgattt accaggtacc attactttaa taagaaaagg tatagttact attgtaatca
     3301 taggaagtat aagtaggtaa gaccaatgaa tataatgtgc tattattccc cctattgaag
     3361 gacctaaccc ttcacctaaa gctacaattg atcctataaa accaaaggct ttgccttgtt
     3421 tttttcttgt aatatttcta gctacaacca ccataatcag tgaagggaat gcagcagatc
     3481 ctactccttg tactaaccta ccaaaaatca aaataaaaaa gtgattgtga ccaataaaag
     3541 caatcaatga accaagacag ctcaaactaa taccaataat taacaatttt tttatattta
     3601 tataatcaga taattttcca tatactgctg ttcctatcga aaaagttaac atatatgcag
     3661 tgtttaccca gtttgtaatt ccaggagtag tattaaaatg atttgcaata tcaggtaaag
     3721 aaacatttaa aaccatttca tttaatacac taaaaaatga aagaatacaa agccaaaata
     3781 aaacgctata aaacaaacct ttaaattttt tatataaact aaacaatttt acctctttac
     3841 ttgaggtgac taaagtttat aggggtaaat tattctttca cctcatataa attccccaaa
     3901 tttttaaata gacacttcat aaaaaaatcc tcctaaatta gaatagttta aatatatcat
     3961 ttttgttcag taatattaat atgtacacta tttccaaaat ttaaattcat gttgccaaaa
     4021 atcgatttgt ttttgcaatt gtttttcgtt cgcttttaaa gtcgatttca ttaagtccgt
     4081 taaatcgact ggcgaaattt cttttatcaa attcttatat tggatctgct gcattaatga
     4141 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     4201 actgactcgc tgcgctcggt cgttcggctg cggcgcgccg tcggcttaaa ccagttttcg
     4261 ttggtgcgaa aaaagagtgt cttgtgacac tcttaaattc aaaatctatc ggtcagattt
     4321 ataccgattt gattttatat attcttgaat aacatacgcc gagttatcac ataaaagcgg
     4381 gaaccaatca tcaaatttaa acttcattgc ataatccatt aaactcttaa attctacgat
     4441 tccttgttca tcaataaact caatcatttc tttaattaat ttatatctat ctgttgttgt
     4501 tttctttaat aattcatcaa catctacacc gccataaact atcatatctt ctttttgata
     4561 tttaaattta ttaggatcgt ccatgtgaag catatatctc acaagacctt tcacacttcc
     4621 tgcaatctgc ggaatagtcg cattcaattc ttctgtaatt atttttatct gttcataaga
     4681 tttattaccc tcatacatca ctagaatatg ataatgctct tttttcatcc tatcttctgt
     4741 atcagtatcc ctatcatgta atggagacac tacaaattga atgtgtaact cttttaaata
     4801 ctctaaccac tcggcttttg ctgattctgg atataaaaca aatgtccaat tacgtcctct
     4861 tgaatttttc ttgttttcag tttcttttat tacattttcg ctcatgatat aataacggtg
     4921 ctaatacatt taacaaaatt tagtcataga taggcagcat gccagtgctg tctatctttt
     4981 tttgtttaaa atgcaccgta ttcctccttt gcatattttt ttattagaat accggttgca
     5041 tctgatttgc taatattata tttttctttg attctattta atatctcatt ttcttctgtt
     5101 gtaagtctta aagtaacagc aacttttttc tcttcttttc tatctacaac catcactgta
     5161 cctcccaaca tctgtttttt tcactttaac ataaaaaaca accttttaac attaaaaacc
     5221 caatatttat ttatttgttt ggacaatgga caatggacac ctagggggga ggtcgtagta
     5281 cccccctatg ttttctcccc taaataaccc caaaaatcta agaaaaaaag acctcaaaaa
     5341 ggtctttaat taacatctca aatttcgcat ttattccaat ttcctttttg cgtgtggcgc
     5401 gccggcacgc gtatcgatgc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5461 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5521 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5581 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5641 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5701 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5761 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5821 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5881 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5941 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6001 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6061 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6121 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6181 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6241 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6301 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6361 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6421 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6481 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6541 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6601 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6661 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6721 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6781 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6841 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6901 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6961 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7021 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7081 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7141 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7201 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7261 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7321 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
