LOCUS       V003685                 6663 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003685
VERSION     V003685
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6663)
  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 6663)
  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 6663)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6663)
  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..6663
                     /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(2410..3141)
                     /gene="ermC"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from
                     Staphylococcus aureus. Accession#: P02979"
     rep_origin      complement(3539..4703)
                     /direction=LEFT
                     /note="thermosensitive origin of replication; for
                     replication in Staphylococcus aureus; pE194ts"
     rep_origin      complement(4844..5432)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5606..6463)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6464..6568)
                     /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 gtagatctca caaaaaatag gcacacgaaa aacaagttaa gggatgcagt ttatgcatcc
     2401 cttaacttac ttattaaata atttatagct attgaaaaga gataagaatt gttcaaagct
     2461 aatattgttt aaatcgtcaa ttcctgcatg ttttaaggaa ttgttaaatt gattttttgt
     2521 aaatattttc ttgtattctt tgttaaccca tttcataacg aaataattat acttttgttt
     2581 atctttgtgt gatattcttg attttttcct acttaatctg ataagtgagc tattcacttt
     2641 aggtttagga tgaaaatatt ctcttggaac catacttaat atagaaatat caacttctgc
     2701 cattaaaagt aatgccaatg agcgttttgt atttaataat cttttagcaa acccgtattc
     2761 cacgattaaa taaatctcat tagctatact atcaaaaaca attttgcgta ttatatccgt
     2821 acttatgtta taaggtatat taccatatat tttataggat tggtttttag gaaatttaaa
     2881 ctgcaatata tccttgttta aaacttggaa attatcgtga tcaacaagtt tattttctgt
     2941 agttttgcat aatttatggt ctatttcaat ggcagttacg aaattacacc tctttactaa
     3001 ttcaagggta aaatggcctt ttcctgagcc gatttcaaag atattatcat gttcatttaa
     3061 tcttatattt gtcattattt tatctatatt atgttttgaa gtaataaagt tttgactgtg
     3121 ttttatattt ttctcgttca ttataaccct ctttaatttg gttatatgaa ttttgcttat
     3181 taacgattca ttataaccac ttattttttg tttggttgat aatgaactgt gctgattaca
     3241 aaaatactaa aaatgcccat attttttcct ccttataaaa ttagtataat tatagcacaa
     3301 gctctgataa atatgaacat gatgagtgat cgttaaattt atactgcaat cggatgcgat
     3361 tattgaataa aagatatgag agatttatct aatttctttt ttcttgtaaa aaaagaaagt
     3421 tcttaaagga tctgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3481 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3541 gcgccgtcgg cttaaaccag ttttcgttgg tgcgaaaaaa gagtgtcttg tgacactctt
     3601 aaattcaaaa tctatcggtc agatttatac cgatttgatt ttatatattc ttgaataaca
     3661 tacgccgagt tatcacataa aagcgggaac caatcatcaa atttaaactt cattgcataa
     3721 tccattaaac tcttaaattc tacgattcct tgttcatcaa taaactcaat catttcttta
     3781 attaatttat atctatctgt tgttgttttc tttaataatt catcaacatc tacaccgcca
     3841 taaactatca tatcttcttt ttgatattta aatttattag gatcgtccat gtgaagcata
     3901 tatctcacaa gacctttcac acttcctgca atctgcggaa tagtcgcatt caattcttct
     3961 gtaattattt ttatctgttc ataagattta ttaccctcat acatcactag aatatgataa
     4021 tgctcttttt tcatcctatc ttctgtatca gtatccctat catgtaatgg agacactaca
     4081 aattgaatgt gtaactcttt taaatactct aaccactcgg cttttgctga ttctggatat
     4141 aaaacaaatg tccaattacg tcctcttgaa tttttcttgt tttcagtttc ttttattaca
     4201 ttttcgctca tgatataata acggtgctaa tacatttaac aaaatttagt catagatagg
     4261 cagcatgcca gtgctgtcta tctttttttg tttaaaatgc accgtattcc tcctttgcat
     4321 atttttttat tagaataccg gttgcatctg atttgctaat attatatttt tctttgattc
     4381 tatttaatat ctcattttct tctgttgtaa gtcttaaagt aacagcaact tttttctctt
     4441 cttttctatc tacaaccatc actgtacctc ccaacatctg tttttttcac tttaacataa
     4501 aaaacaacct tttaacatta aaaacccaat atttatttat ttgtttggac aatggacaat
     4561 ggacacctag gggggaggtc gtagtacccc cctatgtttt ctcccctaaa taaccccaaa
     4621 aatctaagaa aaaaagacct caaaaaggtc tttaattaac atctcaaatt tcgcatttat
     4681 tccaatttcc tttttgcgtg tggcgcgccg gcacgcgtat cgatgctcac tcaaaggcgg
     4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5281 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5401 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5461 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5641 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5701 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5761 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5821 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5881 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5941 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6001 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6061 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6121 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6181 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6241 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6301 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6361 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6421 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6481 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6541 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     6601 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6661 gtc
//
