LOCUS       Exported                4128 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p34E-Km, complete sequence.
ACCESSION   KX485327
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  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 4128)
  AUTHORS   Shastri S, Spiewak HL, Pereira T, Sofoluwe AO, Eidsvaag VA, Bull EH,
            Asghar AH, Thomas MS.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUN-2016) Department of Infection, Immunity & 
            Cardiovascular Disease, The University of Sheffield, The Medical 
            School, Beech Hill Road, Sheffield, South Yorkshire S10 2RX, UK
REFERENCE   3  (bases 1 to 4128)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4128
                     /organism="Cloning vector p34E-Km"
                     /mol_type="other DNA"
                     /db_xref="taxon:1918673"
     regulatory      786..890
                     /regulatory_class="promoter"
                     /note="bla promoter"
     promoter        786..890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             891..1751
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers resistance to ampicillin"
                     /protein_id="APD15311.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            891..1751
                     /gene="bla"
                     /label=bla
     rep_origin      1922..2510
                     /note="oriR; ColE1/pMB1/pBR322/pUC origin of replication"
     regulatory      2762..2780
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        2762..2780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2789..2845
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     CDS             complement(2881..3675)
                     /codon_start=1
                     /gene="aphA2"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA2
                     /note="confers resistance to kanamycin"
                     /protein_id="APD15312.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2881..3675)
                     /gene="aphA2"
                     /label=aphA2
     regulatory      complement(3779..3807)
                     /regulatory_class="promoter"
                     /note="aphA2 promoter"
     misc_feature    4049..4105
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      complement(4110..4128)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     promoter        complement(4110..4128)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"