LOCUS       V003226                 7502 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003226
VERSION     V003226
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7502)
  AUTHORS   Shastri S, Spiewak HL, Sofoluwe A, Eidsvaag VA, Asghar AH, Pereira
            T, Bull EH, Butt AT, Thomas MS.
  TITLE     An efficient system for the generation of marked genetic mutants in
            members of the genus Burkholderia
  JOURNAL   Plasmid (2016) In press
   PUBMED   27825973
REFERENCE   2  (bases 1 to 7502)
  AUTHORS   Shastri S, Spiewak HL, Pereira T, Sofoluwe AO, Eidsvaag VA, Bull EH,
            Asghar AH, Thomas MS.
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7502)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7502)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid
            (2016) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (30-JUN-2016) Department of Infection, Immunity "
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1830..2468
                     /gene="cmlA"
                     /label="Chloramphenicol acetyltransferase 2"
                     /note="Chloramphenicol acetyltransferase 2 from Escherichia
                     coli. Accession#: P22615"
     misc_feature    3835..3853
                     /label="O end"
                     /note="O end"
     protein_bind    complement(3865..3881)
                     /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)."
     rep_origin      complement(3916..4304)
                     /direction=LEFT
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(4925..5293)
                     /label="traJ"
                     /note="oriT-recognizing protein"
     oriT            complement(5326..5435)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(6516..7373)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7374..7478)
                     /label="AmpR promoter"
ORIGIN
        1 tgcagatctg gccgcctagg ccgaattccc ggggatccgg tgattgattg agcaagcttt
       61 atgcttgtaa accgttttgt gaaaaaattt ttaaaataaa aaaggggacc tctagggtcc
      121 ccaattaatt agtaatataa tctattaaag gtcattcaaa aggtcatcca ccggatcaat
      181 tcccctgctc gcgcaggctg ggtgccaagc tctcgggtaa catcaaggcc cgatccttgg
      241 agcccttgcc ctcccgcacg atgatcgtgc cgtgatcgaa atccagatcc ttgacccgca
      301 gttgcaaacc ctcactgatc cgcacatcgg gatcacttgt tcggtgaggt agcagtcgaa
      361 gtcgcattgc agcgcacgct tgaggaacgg ggtgaggctg gtgccttcgg tggagcgcca
      421 ccgcgccatg acctggatgc aacgcctcaa gcgagtcttc aatattgata ttgaagtctg
      481 cgaacactgc ggcggtcacg tcaaagtgat tgccagcatc gaagatccga aggtcattga
      541 gcagattctc aagcatctga aacagaaaac agccaaggcg aatgccgcca agcagcgtga
      601 gctgccacca gaacgagcgc cgccactgac tcccagcctg ttcgatccat cacagagtcg
      661 tctctttgac tgacgacccc aaatccaaca ctgctcaaca ctgccaactt ttaaacgggg
      721 cggtggggca gtttgtatct ctcgagctat caggctagag attttaccgc caaatcgaac
      781 cttattagag cggtttaggc tggaccggca gttaaaattg gggcttgagc ggtaaacgag
      841 tgagggaatt tcaggtaaga tacttcggat gaggagcaaa aaggtggttt atacttccta
      901 tacccttgcc agcatcgaag atccgaaggt cattgagcag attctcaagc atctgaaaca
      961 gaaaacagcc aaggcgaatg ccgccaagca gcgtgagctg ccaccagaac gagcgccgcc
     1021 actgactccc agcctgttcg atccatcaca gagtcgtctc tttgactgac gaccccaaat
     1081 ccaacactgc tcaacactgc caacttttaa acggggcggt ggggcagttt gtatctctcg
     1141 agctatcagg ctagagattt taccgccaaa tcgaacctta ttagagcggt ttaggctgga
     1201 ccggcagtta aaattggggc ttgagcggta aacgagtgag ggaatttcag gtaagatact
     1261 tcggatgagg agcaaaaagg tggtttatac ttcctatacc caaaaaggtg gtttatactt
     1321 cctatacccc ggttatccgg tggttttctg tttgcggaat aaaacagatt agtgacagtt
     1381 ttctcagaat ggcgtgttcc ggtgacttct gttttcgtgt gctgcatccg gaagtgtcag
     1441 gcccgccgga tatgaaaaag gcctccggag agtaccgtga ctttattttc ggcacaaata
     1501 caggggtcga tgggataaat acggcgatag tttcctgacg gatgatccgt atgtaccggc
     1561 ggaagacaag ctgcaaacct gtcagatgga gattgattta atggcggatg tgctgagagc
     1621 accgccccgt gaatccgcag aactgatccg ctatgtgttt gcggatgatt ggccggaata
     1681 aataaagccg ggcttaatac agattaagcc cgtataggta ttattactga ataccaaaca
     1741 gcttacggag gacggaatgt tacccattga gacaaccaga ctgccttctg attattaata
     1801 tttttcacta ttaatcagaa ggaataacca tgaattttac ccggattgac ctgaatacct
     1861 ggaatcgcag ggaacacttt gccctttatc gtcagcagat taaatgcgga ttcagcctga
     1921 ccaccaaact cgatattacc gctttgcgta ccgcactggc ggagacaggt tataagtttt
     1981 atccgctgat gatttacctg atctcccggg ctgttaatca gtttccggag ttccggatgg
     2041 cactgaaaga caatgaactt atttactggg accagtcaga cccggtcttt actgtctttc
     2101 ataaagaaac cgaaacattc tctgcactgt cctgccgtta ttttccggat ctcagtgagt
     2161 ttatggcagg ttataatgcg gtaacggcag aatatcagca tgataccaga ttgtttccgc
     2221 agggaaattt accggagaat cacctgaata tatcatcatt accgtgggtg agttttgacg
     2281 gatttaacct gaacatcacc ggaaatgatg attattttgc cccggttttt acgatggcaa
     2341 agtttcagca ggaaggtgac cgcgtattat tacctgtttc tgtacaggtt catcatgcag
     2401 tatgtgatgg ctttcatgca gcacggttta ttaatacact tcagctgatg tgtgataaca
     2461 tactgaaata aattaattaa ttctgtattt aagccaccgt atccggcaga atggtggctt
     2521 ttttttatat tttaaccgta atctgtaatt tcgtttcaga ctggttcagg atcacgttac
     2581 gataagtccc ccgcagttgg taataccctt aataaaaaga aacagtcaaa gactacagca
     2641 ataataataa agttaagcag ttaacaataa tattaacaac accagatgca gttataataa
     2701 tagtatttaa gacaccagaa agactgctgc gacagttatt ttgaacaaca ccaaaatgcc
     2761 gtaaaggcag tagtaacaac accagtgaaa gacatcacga tagcatagtg atatgcctga
     2821 gtgtgtgtaa ttaaacaata aataaaccgc catatataac agaaagatag tattctgaat
     2881 ggcatgcttt tctgttcagt ataaacatat catcccggtt ggtataagga tgatatataa
     2941 taagttaagc tgaacacata tttattttgg ttttatttta caaataaagt aagacgatcc
     3001 gttaagtcaa agcggggtat atttattata ccctgctttt ttatttgtcc gccgggcgcg
     3061 gataatggat cagattatgc agtgtcacaa tggccttacc gggattggcg taagcgtgcg
     3121 ggatatccgc atggaagcgc aggattcccc ggcagaaacg gtgtgccact catcccccag
     3181 ccgcagttgt aatgcgcctt ccagtacaat gacatgttct ctggttctga aatccatccc
     3241 tgtcggtgtt gcttatgcag tctggtcggg actcggcgtc gtcataatta cagccattgc
     3301 ctggttgctt catgggcaaa agctcgcgca gatcagttgg aagaatttgt ccactacgtg
     3361 aaaggcgaga tcaccaaggt agtcggcaaa taatgtctaa caattcgttc aagccgacgc
     3421 cgcttcgcgg cgcggcttaa ctcaagcgtt agatgcacta agcacataat tgctcacagc
     3481 caaactatca ggtcaagtct gcttttatta tttttaagcg tgcataataa gccctacaca
     3541 aattgggaga tatatcatga atggctggct ttttcttgtt atcgcaatag ttggcgaagt
     3601 aatcgcaaca tccgcattaa aatctagcga gggctttact aagctgatcc ggtggatgac
     3661 cttttgaatg acctttaata gattatatta ctaattaatt ggggacccta gaggtcccct
     3721 tttttatttt aaaaattttt tcacaaaacg gtttacaagc ataaagcttg ctcaatcaat
     3781 caccggatcc ccgggaattc gtcgacaagc ttcggccgcc taggccgcgg ccgcacttgt
     3841 gtataagagt cagaattcca caaattgtta tccgctcaca attccacatg tggaattcca
     3901 catgtggaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga
     3961 ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct
     4021 taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg
     4081 ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga
     4141 aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt
     4201 tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag
     4261 cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcctctac gccggacgca
     4321 tcgtggccgg atccagccga ccaggctttc cacgcccgcg tgccgctcca tgtcgttcgc
     4381 gcggttctcg gaaacgcgct gccgcgtttc gtgattgtca cgctcaagcc cgtagtcccg
     4441 ttcgagcgtc gcgcagaggt cagcgagggc gcggtaggcc cgatacggct catggatggt
     4501 gtttcgggtc gggtgaatct tgttgatggc gatatggatg tgcaggttgt cggtgtcgtg
     4561 atgcacggca ctgacgcgct gatgctcggc gaagccaagc ccagcgcaga tgcggtcctc
     4621 aatcgcgcgc aacgtctccg cgtcgggctt ctctcccgcg cggaagctaa ccagcaggtg
     4681 ataggtcttg tcggcctcgg aacgggtgtt gccgtgctgg gtcgccatca cctcggccat
     4741 gacagcgggc agggtgtttg cctcgcagtt cgtgacgcgc acgtgaccca ggcgctcggt
     4801 cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga
     4861 gcgcatgggg acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa
     4921 gtcatggctc tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct
     4981 tctcttcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg
     5041 ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc
     5101 gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga
     5161 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
     5221 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
     5281 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
     5341 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     5401 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
     5461 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
     5521 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag cgctgcttcc
     5581 ctgctgtttt gtggaatatc taccgactgg aaacaggcaa atgcaggaaa ttactgaact
     5641 gaggggacag gcgagagacg atgccaaaga gctacaccga cgagctggcc gagtgggttg
     5701 aatcccgcgc ggccaagaag cgccggcgtg atgaggctgc ggttgcgttc ctggcggtga
     5761 gggcggatgt cgaggcggcg ttagcgtccg gctatgcgct cgtcaccatt tgggagcaca
     5821 tgcgggaaac ggggaaggtc aagttctcct acgagacgtt ccgctcgcac gccaggcggc
     5881 acatcaaggc caagcccgcc gatgtgcccg caccgcaggc caaggctgcg gaacccgcgc
     5941 cggcacccaa gacgccggag ccacggcggc cgaagcaggg gggcaaggct gaaaagccgg
     6001 cccccgctgc ggccccgacc ggcttcacct tcaacccaac accggacaaa aaggatcctc
     6061 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     6121 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     6181 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
     6241 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6301 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6361 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     6421 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca taaatcaatc
     6481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     6841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     7141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7501 cc
//
