LOCUS       Exported                4488 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gene replacement vector pBAKA, complete sequence.
ACCESSION   EU277854
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4488)
  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 4488)
  AUTHORS   Barrett AR, Kang Y, Son MS, Vukovich JM, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-NOV-2007) Microbiology, University of Hawaii Manoa, 
            2538 McCarthy Mall Snyder Hall, Rm. 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 4488)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4488
                     /organism="Gene replacement vector pBAKA"
                     /mol_type="other DNA"
                     /db_xref="taxon:495166"
     oriT            20..129
                     /note="incP origin of transfer"
     regulatory      complement(550..875)
                     /regulatory_class="terminator"
     terminator      550..577
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      669..755
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(911..1195)
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ alpha peptide"
                     /label=lacZ
                     /protein_id="ABY75256.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(911..1195)
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     1105..1121
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1125..1181
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1191..1207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1215..1231
                     /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(1239..1269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1284..1305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1528..2146
     rep_origin      complement(1593..2181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2266..3381)
                     /codon_start=1
                     /gene="asd"
                     /product="aspartate semialdehyde dehydrogenase"
                     /label=asd
                     /note="Asd"
                     /protein_id="ABY75257.1"
                     /translation="MKRVGLIGWRGMVGSVLIQRMLEERDFDLIEPVFFTTSNVGAQAP
                     EVDKDIAPLKDAYSIDELKTLDVILTCQGGDYTSEVFPKLREAGWQGYWIDAASSLRME
                     DDAVIVLDPVNRKVIDQALDAGTRNYIGGNCTVSLMLMALGGLFDAGLVEWMSAMTYQA
                     ASGAGAQNMRDLLKQMGAAHASVADDLANPASAILDIDRKVAETLRSEAFPTEHFGAPL
                     GGSLIPWIDKELSQRRQSREEWKAQAETNKILARFKNPIPVDGICVRVGAMRCHSQALT
                     IKLNKDVPLTDIEGLIRQHNPWVKLVPNHREVSVRELTPAAVTGTLSVPVGRLRKLNMV
                     SQYLGAFTVGDQLLWGAAEPLRRMLRILLER"
     gene            complement(2266..3381)
                     /gene="asd"
                     /label=asd
     CDS             complement(3395..4408)
                     /codon_start=1
                     /gene="pheS"
                     /product="phenylalanyl-transfer RNA synthetase alpha 
                     subunit"
                     /label=pheS
                     /note="PheS"
                     /protein_id="ABY75258.1"
                     /translation="MDLDQIVADAQQSFEGAADITTLENEKARFLGKSGALTELLKGLG
                     KLDPETRKTEGARINVAKQQVEAALNARRQALADALLNQRLAAEAIDVTLPGRGAGAGS
                     LHPVMRTWERVEQIFRSIGFDVADGPEIETDWYNFTALNSPENHPARSMQDTFYVDGKD
                     ADGRPLLLRTHTSPMQVRYARMNRPPIKVIAPGRTYRVDSDATHSPMFNQVEGLWIDEN
                     VSFADLKGAYTDFLKKFFERDDILVRFRPSYFPFTEPSAEIDMMFEHGKNAGKWLEISG
                     SGQVHPTVIRNMGLDPERYIGFGFGSGLERLTMLRYGVQDLRLFFENDLRFLRQFA"
     gene            complement(3395..4408)
                     /gene="pheS"
                     /label=pheS
     regulatory      complement(4444..4486)
                     /regulatory_class="promoter"