LOCUS       Exported                7212 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Himar1-delivery and mutagenesis vector pHBurk3, complete sequence.
ACCESSION   EU919403
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7212
                     /organism="Himar1-delivery and mutagenesis vector pHBurk3"
                     /mol_type="other DNA"
                     /db_xref="taxon:580601"
     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
                     /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)."
     CDS             complement(1838..2632)
                     /codon_start=1
                     /gene="npt-II"
                     /product="neomycin phosphotransferase II"
                     /label=npt-II
                     /note="Npt-II; kanamycin resistance protein"
                     /protein_id="ACK37847.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(1838..2632)
                     /gene="npt-II"
                     /label=npt-II
     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      3173..3881
                     /label=ori R6K present on Himar1
                     /note="ori R6K present on Himar1"
     rep_origin      3280..3668
                     /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      4554..5227
                     /label=pUC ori derived from pCR2.1
                     /note="pUC ori derived from pCR2.1"
     rep_origin      4557..5145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5433..5454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5625..6458)
                     /codon_start=1
                     /product="ts replication protein"
                     /label=ts replication protein
                     /note="Rep-ts"
                     /protein_id="ACK37848.1"
                     /translation="MASPPMVYKSNALVEAAYCLSVQEQRIVLASISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMGSTHAIRLYELLTQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKARKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     rep_origin      6459..6859
                     /note="region containing broad-host-range origin of 
                     replication from pRO1600"
     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            6896..7155
     oriT            7040..7148
                     /note="incP origin of transfer"