LOCUS       V003689                 6581 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003689
VERSION     V003689
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6581)
  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 6581)
  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 6581)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6581)
  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..6581
                     /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(2424..3071)
                     /gene="cat"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     rep_origin      complement(3458..4621)
                     /direction=LEFT
                     /note="thermosensitive origin of replication; for
                     replication in Staphylococcus aureus; pE194ts"
     rep_origin      complement(4762..5350)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5524..6381)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6382..6486)
                     /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 gtagatcttc ttcaactaac ggggcaggtt agtgacatta gaaaaccgac tgtaaaaagt
     2401 acagtcggca ttatctcata ttataaaagc cagtcattag gcctatctga caattcctga
     2461 atagagttca taaacaatcc tgcatgataa ccatcacaaa cagaatgatg tacctgtaaa
     2521 gatagcggta aatatattga attaccttta ttaatgaatt ttcctgctgt aataatgggt
     2581 agaaggtaat tactattatt attgatattt aagttaaacc cagtaaatga agtccaagga
     2641 ataatagaaa gagaaaaagc attttcaggt ataggtgttt tgggaaacaa tttccccgaa
     2701 ccattatatt tctctacatc agaaaggtat aaatcataaa actctttgaa gtcattcttt
     2761 acaggagtcc aaataccaga gaatgtttta gatacaccat caaaaattgt ataaagtggc
     2821 tctaacttat cccaataacc taactctccg tcgctattgt aaccagttct aaaagctgta
     2881 tttgagttta tcacccttgt cactaagaaa ataaatgcag ggtaaaattt atatccttcc
     2941 tgttttatgt ttcggtataa aacactaata tcaatttctg tggttatact aaaagtcgtt
     3001 tgttggttca aataatgatt aaatatctct tttctcttcc aattgtctaa atcaatttta
     3061 ttaaagttca tttgatgtgc ctcctaaatt tttatctaaa gtgaatttag gaggcttact
     3121 tgtctgcttt cttcattaga atcaatcctt ttttaaaagt caatattact gtaacataaa
     3181 tatatatttt aaaaatatcc cactttatcc aattttcgtt tgttgaacta atgggtgctt
     3241 tagttgaaga ataaaagacc acattaaaaa atgtggtctt ttgtgttttt ttaaaggatt
     3301 tgagcgtagc gaaaaatcct tttctttctt atcttgagaa tacatggatc tgctgcatta
     3361 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3421 gctcactgac tcgctgcgct cggtcgttcg gctgcggcgc gccgtcggct taaaccagtt
     3481 ttcgttggtg cgaaaaaaga gtgtcttgtg acactcttaa attcaaaatc tatcggtcag
     3541 atttataccg atttgatttt atatattctt gaataacata cgccgagtta tcacataaaa
     3601 gcgggaacca atcatcaaat ttaaacttca ttgcataatc cattaaactc ttaaattcta
     3661 cgattccttg ttcatcaata aactcaatca tttctttaat taatttatat ctatctgttg
     3721 ttgttttctt taataattca tcaacatcta caccgccata aactatcata tcttcttttt
     3781 gatatttaaa tttattagga tcgtccatgt gaagcatata tctcacaaga cctttcacac
     3841 ttcctgcaat ctgcggaata gtcgcattca attcttctgt aattattttt atctgttcat
     3901 aagatttatt accctcatac atcactagaa tatgataatg ctcttttttc atcctatctt
     3961 ctgtatcagt atccctatca tgtaatggag acactacaaa ttgaatgtgt aactctttta
     4021 aatactctaa ccactcggct tttgctgatt ctggatataa aacaaatgtc caattacgtc
     4081 ctcttgaatt tttcttgttt tcagtttctt ttattacatt ttcgctcatg atataataac
     4141 ggtgctaata catttaacaa aatttagtca tagataggca gcatgccagt gctgtctatc
     4201 tttttttgtt taaaatgcac cgtattcctc ctttgcatat ttttttatta gaataccggt
     4261 tgcatctgat ttgctaatat tatatttttc tttgattcta tttaatatct cattttcttc
     4321 tgttgtaagt cttaaagtaa cagcaacttt tttctcttct tttctatcta caaccatcac
     4381 tgtacctccc aacatctgtt tttttcactt taacataaaa aacaaccttt taacattaaa
     4441 aacccaatat ttatttattt gtttggacaa tggacaatgg acacctaggg gggaggtcgt
     4501 agtacccccc tatgttttct cccctaaata accccaaaaa tctaagaaaa aaagacctca
     4561 aaaaggtctt taattaacat ctcaaatttc gcatttattc caatttcctt tttgcgtgtg
     4621 gcgcgccggc acgcgtatcg atgctcactc aaaggcggta atacggttat ccacagaatc
     4681 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4741 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4801 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4861 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4921 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4981 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5041 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5101 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5161 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5221 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5281 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5341 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5401 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5461 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5521 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5581 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5641 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5701 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5761 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5821 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5881 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5941 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6001 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6061 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6121 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6181 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6241 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6301 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6361 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6421 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6481 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     6541 attaacctat aaaaataggc gtatcacgag gccctttcgt c
//
