LOCUS       40924_24148        7244 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Himar1-delivery and mutagenesis vector pHBurk5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7244)
  AUTHORS   Rholl DA, Trunck LA, Schweizer HP.
  TITLE     In vivo Himar1 transposon mutagenesis of Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (24), 7529-7535 (2008)
  PUBMED    18952878
REFERENCE   2  (bases 1 to 7244)
  AUTHORS   Rholl DA, Trunck LA, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JUL-2008) Microbiology, Immunology and Pathology, 
            Colorado State University, 1692 Campus Delivery, Fort Collins, CO 
            80523, USA
REFERENCE   3  (bases 1 to 7244)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7244)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "24"; 
            pages: "7529-7535"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-JUL-2008) Microbiology, Immunology and Pathology, Colorado State
            University, 1692 Campus Delivery, Fort Collins, CO 80523, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(104..1150)
                     /codon_start=1
                     /gene="tnp"
                     /product="transposase"
                     /label=tnp
                     /note="Tnp"
                     /protein_id="ACK37849.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQQRVDDSERCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     gene            complement(104..1150)
                     /gene="tnp"
                     /label=tnp
     repeat_region   1537..1565
     promoter        complement(1669..1699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1717..1764
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1875..2666)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    3052..3099
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     3159..3175
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3312..3700)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     repeat_region   3987..4015
     rep_origin      4589..5177
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5465..5486
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5501..5531
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5539..5555
                     /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)."
     primer_bind     5563..5579
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5660..6490)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      6504..6855
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     oriT            7072..7180
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 ggccgctcga gcatgcatct agtaacggcc gccagtgtgc tggaattctg cagattccgg
       61 tctaacaaag aaaaacacat ttttttgtca aaattcgttt ttattattca acatagttcc
      121 cttcaagagc gatacaacga ttataacgac cttccaattt tttgatacca ttttggtagt
      181 actccttcgg ttttgcctca aaataggcct cagtttcggc gatcacctct tcattgcagc
      241 caaatttttt ccctgcgagc atccttttga ggtctgagaa caagaaaaag tcgctggggg
      301 ccagatctgg agaatacggt gggtggggaa gcaattcgaa gcccaattca tgaatttttg
      361 ccatcgttct caatgacttg tggcacggtg cgttgtcttg gtggaacaac acttttttct
      421 tcttcatgtg gggccgtttt gccgcgattt cgaccttcaa acgctccaat aacgccatat
      481 aatagtcact gttgatggtt tttcccttct caagataatc gataaaaatt attccatgcg
      541 catcccaaaa aacagaggcc attactttgc cagcggactt ttgagtcttt ccacgcttcg
      601 gagacggttc accggtcgct gtccactcag ccgactgtcg attggactca ggagtgtagt
      661 gatggagcca tgtttcatcc attgtcacat atcgacggaa aaactcgggt gtattacgag
      721 ttaacagctg caaacaccgc tcagaatcat caacacgttg ttgtttttgg tcaaatgtga
      781 gctcgcgcgg cacccatttt gcacagagct tccgcatatc caaatattga tgaatgatat
      841 gaccaacacg ttcctttgat atctttaagg cctctgctat ctcgatcaac ttcattttac
      901 ggtcattcaa aatcattttg tggatttttt tgatgttttc gtcggtaacc acctctttcg
      961 ggcgtccact gcgttcaccg tcctccgtgc tcatttcacc acgcttgaat tttgcatacc
     1021 aatcaattat tgttgatttc cctggggcag agtccggaaa ctcattatca agccaagttt
     1081 ttgcttccac cgtatttttt cccttcagaa aacagtattt tatcaaaaca cgaaattcct
     1141 ttttttccat atgtatatct ccttcttact ggtaaagcct atctgctgac cgatgttccg
     1201 gcctgccaga ctggacggtc tagtaacggc cgccagtgtg ctggaattct gcagatttta
     1261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcccgggtt
     1321 agagatctcg atgtcagcga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1501 ttgcccggcg tcaatacggg ataataccgc gccacataac aggttggctg ataagtcccc
     1561 ggtctaacaa agaaaaacac atttttttgt caaaattcgt ttttattatt caacatagtt
     1621 cccttcaaga gcgatacccg agctcgaatt gacgcgaatt cggcttcaca acatacgagc
     1681 cggaagcata aagtgtaaag ccgaattggg gatcttgaag tacctattcc gaagttccta
     1741 ttctctagaa agtataggaa cttcagagcg cttttgaagc tgatgtgctt aaaaacttac
     1801 tcaatggaat aattctagat aattcactag tgattggaaa tgttggatcc tcatactctt
     1861 cctttttcaa ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     1921 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     1981 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     2041 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     2101 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     2161 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     2221 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     2281 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     2341 ggagcaaggt gggatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     2401 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     2461 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     2521 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     2581 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     2641 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     2701 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     2761 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat
     2821 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     2881 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     2941 cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccatgca
     3001 tttgttaatc gaattatcca ttgctgttga caaagggaat caggggatct tgaagttcct
     3061 attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt gaagctaatt
     3121 cgaagccgaa ttcgcgtcct cggtaccggg ccccccctcg aggtcgacgg tatcgataag
     3181 cttgatatcg aattcctgca gcccttaatt aaccccgaaa agtgccacct gacgtctaag
     3241 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3301 ttcaagaatt ctgtcagccg ttaagtgttc ctgtgtcact gaaaattgct ttgagaggct
     3361 ctaagggctt ctcagtgcgt tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa
     3421 agctttaaaa gccttatata ttcttttttt tcttataaaa cttaaaacct tagaggctat
     3481 ttaagttgct gatttatatt aattttattg ttcaaacatg agagcttagt acgtgaaaca
     3541 tgagagctta gtacgttagc catgagagct tagtacgtta gccatgaggg tttagttcgt
     3601 taaacatgag agcttagtac gttaaacatg agagcttagt acgtgaaaca tgagagctta
     3661 gtacgtacta tcaacaggtt gaactgctga tcttcagatc ctctacgccg gacgcatcgt
     3721 ggggatctgg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     3781 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3841 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3901 cagtttgaga tctgggaatc atttgaaggt tggtactata taaaaataat atgcatttaa
     3961 tactagcgac gccatctaga tgtgtcagac cggggactta tcagccaacc tgttagcaga
     4021 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4081 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4141 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4201 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca aatccatcac
     4261 actggcggcc tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4321 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4381 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4441 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4501 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4561 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4621 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4681 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     4741 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4801 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4861 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4921 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4981 attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5041 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5101 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5161 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5221 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5281 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5341 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5401 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     5461 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     5521 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     5581 atgattacgc caagcttggt accgagctcg gatccactag taacggccgc cagtgtgctg
     5641 gaattcggct tggcctctag gccagatcca gcggcatctg ggttagtcga gcgcgggccg
     5701 cttcccatgt ctcaccaggg cgagcctgtt tcgcgatctc agcatctgaa atcttcctgg
     5761 ccttgcgctt cgcaggggcc ttacccaccg ccttggcggg cttcttcggt ccaaaactga
     5821 acaacagatg tgtgaccttg cgcccggtct ttcgctgcgc ccactccacc tgtagcgggc
     5881 tgtgctcgtt gatctgcgtc acggctggat caagcactcg caacttgaag tccttgatcg
     5941 agggataccg gccttccagt tgaaaccact ttcgcagctg gtcaatttct atttcacgct
     6001 ggccgatgct gtcccattgc gtgagcagct cgtaaagcct gatcgcgtgg gtgctgccca
     6061 tcttggccac gtcagccaag gcgtatttgg tgaactgttt ggtgagttcc gtcagatacg
     6121 gcagcatgtc tttggtgaac ctgagttcta cacggccctc accctcccgg tagatgattg
     6181 tttgcaccca gccggtaatc atcacactcg gtcttttccc cttgccattg ggctcttggg
     6241 ttaaccggac ttcccgccgt ttcaggcgca gggccgcttc tttgagctgg ttgtaggaag
     6301 attcgatagg gacacccgcc atcgtcgcta tgtcctccgc cgtcactgaa tacatcactt
     6361 catcggtgac aggctcgctc ctcttcacct ggctaatact ggccagaacg atccgctgtt
     6421 cctgaacact gaggcaatac gcggcctcga ccagggcatt gcttttgtaa accattgggg
     6481 gtgaggccac gttcgacatt ccttgtgtat aaggggacac tgtatcagcg tcccacaata
     6541 caacaaatcc gtccctttac aacaacaaat ccgtcccttc ttaacaacaa atccgtccct
     6601 taatggcaac aaatccgtcc ctttttaaac tctacaggcc acggattacg tggcctgtag
     6661 acgtcctaaa aggtttaaaa gggaaaagga agaaaagggt ggaaacgcaa aaaacgcacc
     6721 actacgtggc cccgttgggg ccgcatttgt gcccctgaag gggcggggga ggcgtctggg
     6781 caatccccgt tttaccagtc ccctatcgcc gcctgagagg gcgcaggaag cgagtaatca
     6841 gggtatcgag gcggattcac ccttggcgtc caaccagcgg caccagcggc gaattcacga
     6901 aaagccgaat tctgcagatg ggattcctta aggtatactt tccgctgcat aaccctgctt
     6961 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     7021 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     7081 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     7141 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg ataagcttgg cgtaatcatg
     7201 gtcatcatat ggaattatcc atcacactgg cggccgctcg agca
//
