LOCUS       Exported                9633 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKaKa1, complete sequence.
ACCESSION   GU074524
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9633)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Knock-out and pull-out recombineering protocols for natural 
            transformable Burkholderia thailandensis and Burkholderia 
            pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9633)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-OCT-2009) Molecular Biosciences and Biological 
            Engineering, University of Hawaii at Manoa, 2538 McCarthy Mall, 
            Snyder Hall 207, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 9633)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9633
                     /organism="Cloning vector pKaKa1"
                     /mol_type="other DNA"
                     /db_xref="taxon:908464"
     rep_origin      885..1702
                     /label=broad-host-range origin of replication
                     /note="broad-host-range origin of replication"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2455
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="required for plasmid replication"
                     /protein_id="ADO63838.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            1793..2455
                     /gene="rep"
                     /label=rep
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3254..4267)
                     /codon_start=1
                     /gene="pheS"
                     /product="mutant phenylalanine tRNA synthetase alpha 
                     subunit"
                     /label=pheS
                     /note="incorporates phenylalanine and chlorinated 
                     phenylalanine"
                     /protein_id="ADO63839.1"
                     /translation="MDLDQIVADAQQSFEGAADITTLENEKARFLGKSGALTELLKGLG
                     KLDPETRKTEGARINVAKQQVEAALNARRQALADALLNQRLAAEAIDVTLPGRGAGAGS
                     LHPVMRTWERVEQIFRSIGFDVADGPEIETDWYNFTALNSPENHPARSMQDTFYVDGKD
                     ADGRPLLLRTHTSPMQVRYARMNRPPIKVIAPGRTYRVDSDATHSPMFNQVEGLWIDEN
                     VSFADLKGAYTDFLKKFFERDDILVRFRPSYFPFTEPSAEIDMMFEHGKNAGKWLEISG
                     SGQVHPTVIRNMGLDPERYIGFGFGSGLERLTMLRYGVQDLRLFFENDLRFLRQFA"
     gene            complement(3254..4267)
                     /gene="pheS"
                     /label=pheS
     terminator      4381..4625
                     /label=-lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(4626..5306)
                     /codon_start=1
                     /gene="exo"
                     /product="exonuclease"
                     /label=exo
                     /note="for lambda-red recombination"
                     /protein_id="ADO63840.1"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSQDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     gene            complement(4626..5306)
                     /gene="exo"
                     /label=exo
     CDS             complement(5303..6088)
                     /codon_start=1
                     /gene="beta"
                     /product="beta protein"
                     /label=beta
                     /note="for lambda-red recombination"
                     /protein_id="ADO63841.1"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPIRVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     gene            complement(5303..6088)
                     /gene="beta"
                     /label=beta
     CDS             complement(6094..6510)
                     /codon_start=1
                     /gene="gam"
                     /product="gam protein"
                     /label=gam
                     /note="for lambda-red recombination"
                     /protein_id="ADO63842.1"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     gene            complement(6094..6510)
                     /gene="gam"
                     /label=gam
     promoter        complement(6554..6838)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      complement(6562..6589)
                     /regulatory_class="promoter"
                     /note="ParaB; araB promoter"
     regulatory      6686..6714
                     /regulatory_class="promoter"
                     /note="Pc; arac promoter"
     CDS             6865..7743
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="repressor of ParaB"
                     /protein_id="ADO63843.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            6865..7743
                     /gene="araC"
                     /label=araC
     primer_bind     complement(7778..7794)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7824..7842)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7863..7879)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7887..7903
                     /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(7911..7941)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7956..7977
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(8382..9176)
                     /codon_start=1
                     /gene="kmr"
                     /product="kanamycin resistance"
                     /label=kmr
                     /protein_id="ADO63844.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(8382..9176)
                     /gene="kmr"
                     /label=kmr
     CDS             complement(9434..799)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /label=mob
                     /note="required for plasmid mobilization"
                     /protein_id="ADO63845.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"
     gene            complement(9434..799)
                     /gene="mob"
                     /label=mob