LOCUS       40924_24143        7212 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Himar1-delivery and mutagenesis vector pHBurk3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7212)
  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 7212)
  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 7212)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7212)
  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..7212
                     /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="ACK37846.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQQRVDDSERCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     gene            complement(104..1150)
                     /gene="tnp"
                     /label=tnp
     repeat_region   1537..1565
                     /label=from Himar1
                     /note="from Himar1"
     protein_bind    1684..1731
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1841..2632)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    3018..3065
                     /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     3127..3143
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3280..3668)
                     /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   3955..3983
                     /label=from Himar1
                     /note="from Himar1"
     rep_origin      4557..5145
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5433..5454
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5469..5499
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5507..5523
                     /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     5531..5547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5628..6458)
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYCLSVQEQRIVLASISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMGSTHAIRLYELLTQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKARKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      6472..6823
                     /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            7040..7148
                     /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 gacgcggggt accgagctcg aattggggat
     1681 cttgaagtac ctattccgaa gttcctattc tctagaaagt ataggaactt cagagcgctt
     1741 ttgaagctga tgtgcttaaa aacttactca atggaataat tctagataat tcactagtga
     1801 tggaaatgtt ggatcctcat actcttcctt tttcaattca gaagaactcg tcaagaaggc
     1861 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     1921 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     1981 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     2041 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     2101 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     2161 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     2221 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     2281 cagccatgat ggatactttc tcggcaggag caaggtggga tgacaggaga tcctgccccg
     2341 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     2401 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     2461 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     2521 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     2581 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     2641 atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca
     2701 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     2761 gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     2821 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     2881 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtgttccgct
     2941 tcctttagca gcccttgcgc catgcatttg ttaatcgaat tatccattgc tgttgacaaa
     3001 gggaatcagg ggatcttgaa gttcctattc cgaagttcct attctctaga aagtatagga
     3061 acttcagagc gcttttgaag ctaattcgag ctcggtaccc cgcgtcctcg gtaccgggcc
     3121 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccttaattaa
     3181 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3241 aataggcgta tcacgaggcc ctttcgtctt caagaattct gtcagccgtt aagtgttcct
     3301 gtgtcactga aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc
     3361 ttgttgtcca caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc
     3421 ttataaaact taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt
     3481 caaacatgag agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta
     3541 gtacgttagc catgagggtt tagttcgtta aacatgagag cttagtacgt taaacatgag
     3601 agcttagtac gtgaaacatg agagcttagt acgtactatc aacaggttga actgctgatc
     3661 ttcagatcct ctacgccgga cgcatcgtgg ggatctggtt ccgcgcacat ttccccgaaa
     3721 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3841 tagaccgaga tagggttgag tgttgttcca gtttgagatc tgggaatcat ttgaaggttg
     3901 gtactatata aaaataatat gcatttaata ctagcgacgc catctagatg tgtcagaccg
     3961 gggacttatc agccaacctg ttagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4201 catactcttc ctttttcaaa tccatcacac tggcggcctg ggtctcgcgg tatcattgca
     4261 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4321 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4381 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4441 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4501 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4681 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     4741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4921 accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga
     4981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5161 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     5221 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     5281 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     5341 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     5401 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5461 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5521 caatttcaca caggaaacag ctatgaccat gattacgcca agcttggtac cgagctcgga
     5581 tccactagta acggccgcca gtgtgctgga attcggcttg gcctctaggc cagatccagc
     5641 ggcatctggg ttagtcgagc gcgggccgct tcccatgtct caccagggcg agcctgtttc
     5701 gcgatctcag catctgaaat cttcctggcc ttgcgcttcg caggggcctt acccaccgcc
     5761 ttggcgggct tcttcggtcc aaaactgaac aacagatgtg tgaccttgcg cccggtcttt
     5821 cgctgcgccc actccacctg tagcgggctg tgctcgttga tctgcgtcac ggctggatca
     5881 agcactcgca acttgaagtc cttgatcgag ggataccggc cttccagttg aaaccacttt
     5941 cgcagctggt caatttctat ttcacgctgg ccgatgctgt cccattgcgt gagcagctcg
     6001 taaagcctga tcgcgtgggt gctgcccatc ttggccacgt cagccaaggc gtatttggtg
     6061 aactgtttgg tgagttccgt cagatacggc agcatgtctt tggtgaacct gagttctaca
     6121 cggccctcac cctcccggta gatgattgtt tgcacccagc cggtaatcat cacactcggt
     6181 cttttcccct tgccattggg ctcttgggtt aaccggactt cccgccgttt caggcgcagg
     6241 gccgcttctt tgagctggtt gtaggaagat tcgataggga cacccgccat cgtcgctatg
     6301 tcctccgccg tcactgaata catcacttca tcggtgacag gctcgctcct cttcacctgg
     6361 ctaatactgg ccagaacgat ccgctgttcc tgaacactga ggcaatacgc ggcctcgacc
     6421 agggcattgc ttttgtaaac cattgggggt gaggccacgt tcgacattcc ttgtgtataa
     6481 ggggacactg tatcagcgtc ccacaataca acaaatccgt ccctttacaa caacaaatcc
     6541 gtcccttctt aacaacaaat ccgtccctta atggcaacaa atccgtccct ttttaaactc
     6601 tacaggccac ggattacgtg gcctgtagac gtcctaaaag gtttaaaagg gaaaaggaag
     6661 aaaagggtgg aaacgcaaaa aacgcaccac tacgtggccc cgttggggcc gcatttgtgc
     6721 ccctgaaggg gcgggggagg cgtctgggca atccccgttt taccagtccc ctatcgccgc
     6781 ctgagagggc gcaggaagcg agtaatcagg gtatcgaggc ggattcaccc ttggcgtcca
     6841 accagcggca ccagcggcga attcacgaaa agccgaattc tgcagatggg attccttaag
     6901 gtatactttc cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     6961 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     7021 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     7081 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     7141 gctggccgat aagcttggcg taatcatggt catcatatgg aattatccat cacactggcg
     7201 gccgctcgag ca
//
