LOCUS       V003682                 7404 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003682
VERSION     V003682
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7404)
  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 7404)
  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 7404)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7404)
  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..7404
                     /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(5585..6173)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6347..7204)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7205..7309)
                     /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 tttgttctgg aatttctgtg
     4141 tagctttccc cattcttctt catcactcaa taacataaaa actattgctc tatccgcagt
     4201 gcgttggata gttttccaat ctggttgcaa gatatgtaaa tcactaaagc aatcattcca
     4261 gtaatccacc atatctcttt taaattcgat ttctacacgc cataaatgtt cagacataac
     4321 ttcagcatct gcattatctt tacgttcttg ctttttatta taaattctaa taaatctatt
     4381 actatctctc acgccaaaat attttgtttc tggcttacca ttacgaccat aaaaaatagt
     4441 tttcttaact gctttatcag acattgcata gtagtcactc aaatcatctt caaaatcaaa
     4501 ggctaaatct aatcttgtaa aaccgtcatc ttccatgtag cttattatat tttgttttaa
     4561 ccaaatcatt tcatctcgtg taagtttgtt tggattaaat tcaatacgca tattacgtct
     4621 atcccaacta tctgctttca ctttgtcata ttcaatataa actttttctt gaagtgcttt
     4681 agctttaaac tttgtttgaa gaatatccca aagtcttatt tggggctcta cactcataaa
     4741 tttagaaagg gcttgagcgt tgtctcggtt gagatttcca acgatcgtca ttgcatcaaa
     4801 acttaatttt ggatcagata tgcacacagt atgtgcgtcc aaccggctat tagagtagcc
     4861 ggttttagaa aaattgtcta aatcgtgatt ttccaaatga tttgaatgat ttgcatgatt
     4921 gtttttatac ataaaaaatc gactccttaa tctcaatttc gtttaaggaa tcgctcaccc
     4981 aaatatatat cttgatgtat atttaaatat cgtttaatat ctaaatatac aagattataa
     5041 aaacaactca gtgttttttt ctttgaatga tgtcgttcac aaactttggt caggtcgtga
     5101 gcgactcctt tttattttgt tattaatata acactatcaa aagatttggt ctaatcagat
     5161 caagtctttt tttatttaag catttgtatt atctggtaaa cagttaaaac tactaaaaca
     5221 ccaagtacat acttacttgt tataaaattc tcaaaggctt ctctgaattt catttcttca
     5281 actcctttta ttgctttatt ccaatttcct tattggtcgg aacctacagg gctttatgtg
     5341 ttgttgagtt ggtactttct tgggattaat cccaattcaa gtccaaccaa ctcgctaaca
     5401 agttagctaa cacatagccc attccaacca ataagttttc tcggcgcgcc ggcacgcgta
     5461 tcgatgctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6001 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7321 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7381 ggcgtatcac gaggcccttt cgtc
//
