LOCUS       V005257                 9633 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005257
VERSION     V005257
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9633)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Knock-out and pull-out recombineering protocols for natural
            transformable Burkholderia thailandensis and Burkholderia
            pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9633)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-OCT-2009) Molecular Biosciences and Biological
            Engineering, University of Hawaii at Manoa, 2538 McCarthy Mall,
            Snyder Hall 207, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 9633)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9633)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-OCT-2009) Molecular Biosciences and Biological Engineering,
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder Hall 207,
            Honolulu, HI 96822, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     CDS             1793..2452
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     3162..3178
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3188..3206
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3257..4267)
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Burkholderia pseudomallei (strain 1710b). Accession#:
                     Q3JT08"
     terminator      complement(4381..4625)
                     /label="lambda tL3 terminator"
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(4629..5306)
                     /label="Exo"
                     /note="5' to 3' double-stranded DNA exonuclease in the
                     lambda Red system"
     CDS             complement(5306..6088)
                     /label="Beta"
                     /note="single-stranded DNA binding recombinase in the
                     lambda Red system"
     CDS             complement(6097..6510)
                     /label="Gam"
                     /note="inhibitor of the host RecBCD nuclease in the lambda
                     Red system"
     promoter        complement(6554..6838)
                     /label="araBAD promoter"
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite
                     direction (Guzman et al., 1995)"
     CDS             6865..7740
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     primer_bind     complement(7778..7794)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        complement(7824..7842)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7863..7879)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(7887..7903)
                     /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(7911..7941)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7956..7977)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(8385..9176)
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc
     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac
     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     2641 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     2701 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     2761 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     2821 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag
     2881 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     2941 agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3001 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3181 gcgcgcgtaa tacgactcac tatagggcga attggagctc ggtacctcgc gaatgcatct
     3241 agatgaaaga tctttaggcg aactggcgga ggaaccggag gtcattttcg aaaaagagcc
     3301 ggaggtcctg gacaccatac cggagcatgg taaggcgttc caacccggaa ccgaagccga
     3361 agccaatgta acgctccggg tcgaggccca tattccgaat gaccgtgggg tggacctgac
     3421 cgctgccgct gatttccaac cacttaccag cattcttccc atgttcaaac atcatatcaa
     3481 tttccgcgct gggttccgtg aaggggaagt agctcgggcg gaaacggacc aagatgtcat
     3541 ctcgttcgaa aaatttcttg agaaagtcgg tgtaggcacc cttgagatcg gcaaacgaca
     3601 cgttttcatc aatccagagc ccctccacct gattaaacat ggggctatgg gtggcgtccg
     3661 aatcgacccg ataggtccga ccgggagcaa tcaccttaat cgggggtcga ttcatgcggg
     3721 cgtaccgcac ctgcatgggc gaggtatggg tccggaggag gaggggccgc ccatcggcgt
     3781 ccttaccatc cacatagaag gtgtcctgca tggaccgtgc ggggtggttt tcgggcgagt
     3841 tgagcgccgt gaaattatac caatccgtct caatttccgg tccatccgcg acatcgaagc
     3901 caatgctacg gaaaatctgc tccacacgtt cccaggttcg catgacgggg tgcagcgagc
     3961 cggcgcccgc cccgcgaccg gggagggtca catcaatggc ttcagccgcc aggcgctgat
     4021 tgagaagggc atccgccaag gcctgccgcc gggcattcaa ggccgcctcc acctgctgct
     4081 tggccacgtt aatccgggca ccctcggtct tccgggtttc cgggtccagc ttacccagcc
     4141 ccttcaagag ttccgtgagg gcaccgctct tgcccaggaa gcgggccttt tcattttcca
     4201 aggtcgtaat gtcggccgcg ccctcaaagc tctgctgggc atccgccacg atctgatcga
     4261 ggtccatatg cttgtccttt cgaagctcga tccgattgtc agacccggca tgatattccg
     4321 aaaatcccaa gcgagtcaac agctagtact atcggatccc catggattct tcgtctgttt
     4381 ctactggtat tggcacaaac ctgattccaa tttgagcaag gctatgtgcc atctcgatac
     4441 tcgttcttaa ctcaacagaa gatgctttgt gcatacagcc cctcgtttat tatttatctc
     4501 ctcagccagc cgctgtgctt tcagtggatt tcggataaca gaaaggccgg gaaataccca
     4561 gcctcgcttt gtaacggagt agacgaaagt gattgcgcct acccggatat tatcgtgagg
     4621 atgcgtcatc gccattgctc cccaaataca aaaccaattt cagccagtgc ctcgtccatt
     4681 ttttcgatga actccggcac gatctcgtca aaactcgcca tgtacttttc atcccgctca
     4741 atcacgacat aatgcaggcc ttcacgcttc atacgcgggt catagttggc aaagtaccag
     4801 gcattttttc gcgtcaccca catgctgtac tgcacctggg ccatgtaagc tgactttatg
     4861 gcctcgaaac caccgagccg gaacttcatg aaatcctggg aggtaaacgg gcatttcagt
     4921 tcaaggccgt tgccgtcact gcataaacca tcgggagagc aggcggtacg catactttcg
     4981 tcgcgataga tgatcgggga ttcagtaaca ttcacgccgg aagtgaattc aaacagggtt
     5041 ctggcgtcgt tctcgtactg ttttccccag gccagtgctt tagcgttaac ttccggagcc
     5101 acaccggtgc aaacctcagc aagcagggtg tggaagtagg acattttcat gtcaggccac
     5161 ttctttccgg agcggggttt tgctatcacg ttgtgaactt ctgaagcggt gatgacgccg
     5221 agccgtaatt tgtgccacgc atcatccccc tgttcgacag ctctcacatc gatcccggta
     5281 cgctgcagga taatgtccgg tgtcatgctg ccaccttctg ctctgcggct ttctgtttca
     5341 ggaatccaag agcttttact gcttcggcct gtgtcagttc tgacgatgca cgaatgtcgc
     5401 ggcgaaatat ctgggaacag agcggcaata agtcgtcatc ccatgtttta tccagggcga
     5461 tcagcagagt gttaatctcc tgcatggttt catcgttaac cggagtgatg tcgcgttccg
     5521 gctgacgttc tgcagtgtat gcagtatttt cgacaatgcg ctcggcttca tccttgtcat
     5581 agataccagc aaatccgaag gccagacggg cacactgaat catggcttta tgacgtaaca
     5641 tccgtttggg atgcgactgc cacggccccg tgatttctct gccttcgcga gttttgaatg
     5701 gttcgcggcg gcattcatcc atccattcgg taacgcggat cggatgatta cggtccttgc
     5761 ggtaaatccg gcatgtacag gattcattgt cctgctcaaa gtccatgcca tcaaactgct
     5821 ggttttcatt gatgatgcgg gaccagccat caacgcccac caccggaacg atgccattct
     5881 gcttatcagg aaaggcgtaa atttctttcg tccacggatt aaggccgtac tggttggcaa
     5941 cgatcagtaa tgcgatgaac tgcgcatcgc tggcatcacc tttaaatgcc gtctggcgaa
     6001 gagtggtgat cagttcctgt gggtcgacag aatccatgcc gacacgttca gccagcttcc
     6061 cagccagcgt tgcgagtgca gtactcattc gttttatacc tctgaatcaa tatcaacctg
     6121 gtggtgagca atggtttcaa ccatgtaccg gatgtgttct gccatgcgct cctgaaactc
     6181 aacatcgtca tcaaacgcac gggtaatgga ttttttgctg gccccgtggc gttgcaaatg
     6241 atcgatgcat agcgattcaa acaggtgctg gggcaggcct ttttccatgt cgtctgccag
     6301 ttctgcctct ttctcttcac gggcgagctg ctggtagtga cgcgcccagc tctgagcctc
     6361 aagacgatcc tgaatgtaat aagcgttcat ggctgaactc ctgaaatagc tgtgaaaata
     6421 tcgcccgcga aatgccgggc tgattaggaa aacaggaaag ggggttagtg aatgcttttg
     6481 cttgatctca gtttcagtat taatatccat tttttataag cgtcgacgaa ctcgaattcc
     6541 caaaaaaacg ggtatggaga aacagtagag agttgcgata aaaagcgtca ggtaggatcc
     6601 gctaatctta tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca
     6661 atgtggactt ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg
     6721 gtcccgcttt gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag
     6781 agccagtaaa agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttcttct
     6841 ctgaatggcg ggagtatgaa aagtatggct gaagcgcaaa atgatcccct gctgccggga
     6901 tactcgttta atgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc
     6961 gattttttta tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc
     7021 ggtcaggggg tggtgaaaaa tcagggacga gaatttgttt gccgaccggg tgatattttg
     7081 ctgttcccgc caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat
     7141 caccagtggg tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca
     7201 atatttgcca atacggggtt ctttcgcccg gatgaagcgc accagccgca tttcagcgac
     7261 ctgtttgggc aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg
     7321 ataaatctgc ttgagcaatt gttactgcgg cgcatggaag cgattaacga gtcgctccat
     7381 ccaccgatgg ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac
     7441 agcaattttg atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca
     7501 catcttttcc gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc
     7561 agccaggcga agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat
     7621 gttggttttg acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc
     7681 ccgagcgagt tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca
     7741 taataaatgg gctgcaggaa ttcgatatca agcttatcga taccgtcgac ctcgaggggg
     7801 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     7861 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7921 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     7981 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     8041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gcatgcataa aaactgttgt
     8101 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     8161 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggg ggtgggcgaa
     8221 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     8281 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     8341 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     8401 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     8461 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     8521 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     8581 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     8641 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     8701 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     8761 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     8821 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     8881 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     8941 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     9001 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     9061 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     9121 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     9181 aacgatcctc atcctgtctc ttgatcagat cttgatcccc tgcgccatca gatccttggc
     9241 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     9301 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     9361 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     9421 cagtagctga cattcatccc aggtggcact tttcggggaa atgtgcgcgc ccgcgttcct
     9481 gctggcgctg ggcctgtttc tggcgctgga cttcccgctg ttccgtcagc agcttttcgc
     9541 ccacggcctt gatgatcgcg gcggccttgg cctgcatatc ccgattcaac ggccccaggg
     9601 cgtccagaac gggcttcagg cgctcccgaa ggt
//
