LOCUS       V003690                 7038 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003690
VERSION     V003690
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7038)
  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 7038)
  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 7038)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7038)
  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..7038
                     /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(3457..5078)
                     /direction=LEFT
                     /note="thermosensitive origin of replication; for
                     replication in Staphylococcus aureus.; pT181ts"
     rep_origin      complement(5219..5807)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5981..6838)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6839..6943)
                     /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 gcctctttac ttgaggtgac
     3481 taaagtttat aggggtaaat tattctttca cctaatataa attccccaaa tttttaaata
     3541 gacacttcat aaaaaaatcc tcctaaatta gaatagttta aatatatcat ttttgttcag
     3601 taatattaat atgtacgtac actatttcca aaatttaaat tcatgttgcc aaaaatcgat
     3661 ttgtttttgc aattgttttt cgttcgcttt taaagtcgat ttcattaagt ccgttaaatc
     3721 gactggcgaa atttctttta tcaaattctt atattttgtt ctggaatttc tgtgtagctt
     3781 tccccattct tcttcatcac tcaataacat aaaaactatt gctctatccg cagtgcgttg
     3841 gatagttttc caatctggtt gcaagatatg taaatcacta aagcaatcat tccagtaatc
     3901 caccatatct cttttaaatt cgatttctac acgccataaa tgttcagaca taacttcagc
     3961 atctgcatta tctttacgtt cttgcttttt attataaatt ctaataaatc tattactatc
     4021 tctcacgcca aaatattttg tttctggctt accattacga ccataaaaaa tagttttctt
     4081 aactgcttta tcagacattg catagtagtc actcaaatca tcttcaaaat caaaggctaa
     4141 atctaatctt gtaaaaccgt catcttccat gtagcttatt atattttgtt ttaaccaaat
     4201 catttcatct cgtgtaagtt tgtttggatt aaattcaata cgcatattac gtctatccca
     4261 actatctgct ttcactttgt catattcaat ataaactttt tcttgaagtg ctttagcttt
     4321 aaactttgtt tgaagaatat cccaaagtct tatttggggc tctacactca taaatttaga
     4381 aagggcttga gcgttgtctc ggttgagatt tccaacgatc gtcattgcat caaaacttaa
     4441 ttttggatca gatatgcaca cagtatgtgc gtccaaccgg ctattagagt agccggtttt
     4501 agaaaaattg tctaaatcgt gattttccaa atgatttgaa tgatttgcat gattgttttt
     4561 atacataaaa aatcgactcc ttaatctcaa tttcgtttaa ggaatcgctc acccaaatat
     4621 atatcttgat gtatatttaa atatcgttta atatctaaat atacaagatt ataaaaacaa
     4681 ctcagtgttt ttttctttga atgatgtcgt tcacaaactt tggtcaggtc gtgagcgact
     4741 cctttttatt ttgttattaa tataacacta tcaaaagatt tggtctaatc agatcaagtc
     4801 tttttttatt taagcatttg tattatctgg taaacagtta aaactactaa aacaccaagt
     4861 acatacttac ttgttataaa attctcaaag gcttctctga atttcatttc ttcaactcct
     4921 tttattgctt tattccaatt tccttattgg tcggaaccta cagggcttta tgtgttgttg
     4981 agttggtact ttcttgggat taatcccaat tcaagtccaa ccaactcgct aacaagttag
     5041 ctaacacata gcccattcca accaataagt tttctcggcg cgccggcacg cgtatcgatg
     5101 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5641 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6121 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6841 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6961 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     7021 tcacgaggcc ctttcgtc
//
