LOCUS       Exported                5161 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pEX18Km-pheS, complete sequence.
ACCESSION   EU491017
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5161)
  AUTHORS   Barrett AR, Kang Y, Inamasu KS, Son MS, Vukovich JM, Hoang TT.
  TITLE     Genetic tools for allelic replacement in Burkholderia species
  JOURNAL   Appl. Environ. Microbiol. 74 (14), 4498-4508 (2008)
  PUBMED    18502918
REFERENCE   2  (bases 1 to 5161)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Inamasu K, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-FEB-2008) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 5161)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5161
                     /organism="Suicide vector pEX18Km-pheS"
                     /mol_type="other DNA"
                     /db_xref="taxon:508456"
     CDS             complement(315..1328)
                     /codon_start=1
                     /gene="pheS"
                     /product="PheS"
                     /label=pheS
                     /note="tRNA phenylalanyl synthetase alpha subunit"
                     /protein_id="ACA52080.1"
                     /translation="MDLDQIVADAQQSFEGAADITTLENEKARFLGKSGALTELLKGLG
                     KLDPETRKTEGARINVAKQQVEAALNARRQALADALLNQRLAAEAIDVTLPGRGAGAGS
                     LHPVMRTWERVEQIFRSIGFDVADGPEIETDWYNFTALNSPENHPARSMQDTFYVDGKD
                     ADGRPLLLRTHTSPMQVRYARMNRPPIKVIAPGRTYRVDSDATHSPMFNQVEGLWIDEN
                     VSFADLKGAYTDFLKKFFERDDILVRFRPSYFPFTEPSAEIDMMFEHGKNAGKWLEISG
                     SGQVHPTVIRNMGLDPERYIGFGFGSGLERLTMLRYGVQDLRLFFENDLRFLRQFA"
     gene            complement(315..1328)
                     /gene="pheS"
                     /label=pheS
     regulatory      complement(1364..1405)
                     /regulatory_class="promoter"
                     /note="Burkholderia pseudomallei PS12 promoter"
     oriT            1441..1550
                     /note="incP origin of transfer"
     regulatory      complement(1971..2296)
                     /regulatory_class="terminator"
     terminator      1971..1998
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2090..2176
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2332..2616)
                     /codon_start=1
                     /gene="lacZa"
                     /product="LacZa"
                     /label=lacZa
                     /note="LacZ alpha subunit"
                     /protein_id="ACA52079.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(2332..2616)
                     /gene="lacZa"
                     /label=lacZa
     primer_bind     2526..2542
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2546..2602
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2612..2628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2629..2814)
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    2636..2652
                     /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)."
     promoter        complement(2660..2690)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2705..2726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2950..3566
                     /label=Escherichia coli ColE1 origin
                     /note="Escherichia coli ColE1 origin"
     rep_origin      complement(3014..3602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3973..4767)
                     /codon_start=1
                     /gene="kanR"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=kanR
                     /note="kanamycin resistance"
                     /protein_id="ACA52078.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(3973..4767)
                     /gene="kanR"
                     /label=kanR
     promoter        complement(4916..4944)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(4962..5066)
                     /gene="bla"
                     /label=AmpR promoter