LOCUS       V005912                 6601 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005912
VERSION     V005912
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6601)
  AUTHORS   Hamad MA, Skeldon AM, Valvano MA.
  TITLE     Construction of aminoglycoside-sensitive Burkholderia cenocepacia
            strains for use in studies of intracellular bacteria with the
            gentamicin protection assay
  JOURNAL   Appl. Environ. Microbiol. 76 (10), 3170-3176 (2010)
   PUBMED   20348312
REFERENCE   2  (bases 1 to 6601)
  AUTHORS   Hamad MA, Skeldon AM, Valvano MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-DEC-2012) Microbiology and Immunology, Western
            University, 1151 Richmond Street North, London, Ontario N6A 5C1,
            Canada
REFERENCE   3  (bases 1 to 6601)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6601)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2010"; volume: "76"; issue: "10";
            pages: "3170-3176"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (21-DEC-2012) Microbiology and Immunology, Western University, 1151
            Richmond Street North, London, Ontario N6A 5C1, Canada"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            626..735
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             768..1136
                     /label="traJ"
                     /note="oriT-recognizing protein"
     CDS             complement(1102..1527)
                     /codon_start=1
                     /product="TraI"
                     /label="TraI"
                     /protein_id="AGC79933.1"
                     /translation="MNVQVVGVVMHGTDALMLGEAKPSADAVLNRAQRLRVGLLSRAEA
                     NQQVIGLVGLGTGVAVLGRHHLGHDSGQGVCLAVRDAHVTQALGLALLVGDVLHQLREV
                     ALLDGAHGDVLGNHAHPPAVLAAKKVMALPSGGPRPS"
     rep_origin      1757..2145
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     misc_feature    2150..2178
                     /label="multiple cloning site 1"
                     /note="multiple cloning site 1"
     promoter        2194..2222
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             2254..3174
                     /gene="xylE"
                     /label="Metapyrocatechase"
                     /note="Metapyrocatechase from Pseudomonas putida.
                     Accession#: P06622"
     misc_feature    3183..3216
                     /label="multiple cloning site 2"
                     /note="multiple cloning site 2"
     promoter        3945..3973
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     CDS             4299..4532
                     /label="TpR"
                     /note="E. coli plasmid-associated dihydrofolate reductase"
     misc_feature    4538..4555
                     /label="I-SceI recognition site"
                     /note="I-SceI recognition site"
     promoter        4927..5029
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             5030..5668
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label="chloramphenicol acetyltransferase"
                     /protein_id="AGC79930.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
ORIGIN
        1 ggatcctttt tgtccggtgt tgggttgaag gtgaagccgg tcggggccgc agcgggggcc
       61 ggcttttcag ccttgccccc ctgcttcggc cgccgtggct ccggcgtctt gggtgccggc
      121 gcgggttccg cagccttggc ctgcggtgcg ggcacatcgg cgggcttggc cttgatgtgc
      181 cgcctggcgt gcgagcggaa cgtctcgtag gagaacttga ccttccccgt ttcccgcatg
      241 tgctcccaaa tggtgacgag cgcatagccg gacgctaacg ccgcctcgac atccgccctc
      301 accgccagga acgcaaccgc agcctcatca cgccggcgct tcttggccgc gcgggattca
      361 acccactcgg ccagctcgtc ggtgtagctc tttggcatcg tctctcgcct gtcccctcag
      421 ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga tattccacaa aacagcaggg
      481 aagcagcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
      541 tatccatcct ttttcgcacg atatacagga ttaatttgca aagggttcgt gtagactttc
      601 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
      661 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
      721 tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg gctgatgaaa
      781 ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca gacgaacgaa
      841 gagcgattga ggaaaaggcg gcggcggccg gcatgagcct gtcggcctac ctgctggccg
      901 tcggccaggg ctacaaaatc acgggcgtcg tggactatga gcacgtccgc gagctggccc
      961 gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa actctggctc accgacgacc
     1021 cgcgcacggc gcggttcggt gatgccacga tcctcgccct gctggcgaag atcgaagaga
     1081 agcaggacga gcttggcaag gtcatgatgg gcgtggtccg cccgagggca gagccatgac
     1141 ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg ccaagcacgt ccccatgcgc
     1201 tccatcaaga agagcgactt cgcggagctg gtgaagtaca tcaccgacga gcaaggcaag
     1261 accgagcgcc tgggtcacgt gcgcgtcacg aactgcgagg caaacaccct gcccgctgtc
     1321 atggccgagg tgatggcgac ccagcacggc aacacccgtt ccgaggccga caagacctat
     1381 cacctgctgg ttagcttccg cgcgggagag aagcccgacg cggagacgtt gcgcgcgatt
     1441 gaggaccgca tctgcgctgg gcttggcttc gccgagcatc agcgcgtcag tgccgtgcat
     1501 cacgacaccg acaacctgca cattcatatc gccatcaaca agattcaccc gacccgaaac
     1561 accatccatg agccgtatcg ggcctaccgc gccctcgctg acctctgcgc gacgctcgaa
     1621 cgggactacg ggcttgagcg tgacaatcac gaaacgcggc agcgcgtttc cgagaaccgc
     1681 gcgaacgaca tggagcggca cgcgggcgtg gaaagcctgg tcggctggat ccggccacga
     1741 tgcgtccggc gtagaggatc tgaagatcag cagttcaacc tgttgatagt acgtactaag
     1801 ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga
     1861 actaaaccct catggctaac gtactaagct ctcatggcta acgtactaag ctctcatgtt
     1921 tcacgtacta agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag
     1981 cctctaaggt tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta
     2041 aggtttaacg gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc
     2101 tctcaaagca attttgagtg acacaggaac acttaacggc tgacatggga attcgggccc
     2161 cgctagccag atcttcccgg gaagcttgag ctgttgacaa ttaatcatcg gctcgtataa
     2221 tgtgtggaat atcacagaag gagacagaat actatgaaca aaggtgtaat gcgaccgggc
     2281 catgtgcagc tgcgtgtact ggacatgagc aaggccctgg aacactacgt cgagttgctg
     2341 ggcctgatcg agatggaccg tgacgaccag ggccgtgtct atctgaaggc ttggaccgaa
     2401 gtggataagt tttccctggt gctacgcgag gctgacgagc cgggcatgga ttttatgggt
     2461 ttcaaggttg tggatgagga tgctctccgg caactggagc gggatctgat ggcatatggc
     2521 tgtgccgttg agcagctacc cgcaggtgaa ctgaacagtt gtggccggcg cgtgcgcttc
     2581 caggccccct ccgggcatca cttcgagttg tatgcagaca aggaatatac tggaaagtgg
     2641 ggtttgaatg acgtcaatcc cgaggcatgg ccgcgcgatc tgaaaggtat ggcggctgtg
     2701 cgtttcgacc acgccctcat gtatggcgac gaattgccgg cgacctatga cctgttcacc
     2761 aaggtgctcg gtttctatct ggccgaacag gtgctggacg aaaatggcac gcgcgtcgcc
     2821 cagtttctca gtctgtcgac caaggcccac gacgtggcct tcattcacca tccggaaaaa
     2881 ggccgcctcc atcatgtgtc cttccacctc gaaacctggg aagacttgct tcgcgccgcc
     2941 gacctgatct ccatgaccga cacatctatc gatatcggcc caacccgcca cggcctcact
     3001 cacggcaaga ccatctactt cttcgacccg tccggtaacc gcaacgaagt gttctgcggg
     3061 ggagattaca actacccgga ccacaaaccg gtgacctgga ccaccgacca gctgggcaag
     3121 gcgatctttt accacgaccg cattctcaac gaacgattca tgaccgtgct gacctgacct
     3181 gagcggccgc caatattgca tgcggtaccg tctagaagaa gcttgggatc cgtcggggat
     3241 ccggtgattg attgagcaag ctttatgctt gtaaaccgtt ttgtgaaaaa atttttaaaa
     3301 taaaaaaggg gacctctagg gtccccaatt aattagtaat ataatctatt aaaggtcatt
     3361 caaaaggtca tccaccggat cagcttagta aagccctcgc tagattttaa tgcggatgtt
     3421 gcgattactt cgccaactat tgcgataaca agaaaaagcc agcctttcat gatatatctc
     3481 ccaatttgtg tagggcttat tatgcacgct taaaaataat aaaagcagac ttgacctgat
     3541 agtttggctg tgagcaatta tgtgcttagt gcatctaacg cttgagttaa gccgcgccgc
     3601 gaagcggcgt cggcttgaac gaattgttag acattatttg ccgactacct tggtgatctc
     3661 gcctttcacg tagtggacaa attcttccaa ctgatctagc tttaatgcgg tagtttatca
     3721 cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc
     3781 atcctcggca ccgtcaccct ggatgctgta ggcataggct tggttatgcc ggtactgccg
     3841 ggcctcttgc gggattctac ggggtctgac gctcagtgga acgaaatcga tgagctcgca
     3901 cgaacccagt tgacataagc ctgttcggtt cgtaaagaac ccagttgaca taagcctgtt
     3961 cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     4021 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     4081 ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     4141 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag
     4201 gcagccgttg tgctggtgct ttctgatagt tgttgtgggg taggcagtca gagctcgatt
     4261 tgcttgtcgc cataatagat tcacaagaag gattcgacat gggtcaaagt agcgatgaag
     4321 ccaacgctcc cgttgcaggg cagtttgcgc ttcccctgag tgccaccttt ggcttagggg
     4381 atcgcgtacg caagaaatct ggtgccgctt ggcagggtca agtcgtcggt tggtattgca
     4441 caaaactcac tcctgaaggc tatgcggtcg agtccgaatc ccacccaggc tcagtgcaaa
     4501 tttatcctgt ggctgcactt gaacgtgtgg cctaagcatt accctgttat ccctaactgc
     4561 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4621 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4681 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4741 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     4801 atcgataagt ataggaactt cggcgcgcct acctgtgacg gaagatcact tcgcagaata
     4861 aataaatcct ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg
     4921 agacgttgat cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     4981 gggcgtattt tttgagttgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     5041 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     5101 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     5161 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     5221 ccgcctgatg aatgctcatc cggaattacg tatggcaatg aaagacggtg agctggtgat
     5281 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     5341 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     5401 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     5461 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     5521 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     5581 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagatg
     5641 cttaatgaat acaacagtac tgcgatgaat cgatctgcag tcaatggcaa caacgttgcg
     5701 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5761 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5821 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5881 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5941 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaagcgt
     6001 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6061 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6121 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6181 attgatttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     6241 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     6301 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6361 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     6421 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     6481 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     6541 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     6601 a
//
