LOCUS       Exported                4833 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pEX18Ap-pheS, complete sequence.
ACCESSION   EU332346
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4833)
  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 4833)
  AUTHORS   Barrett AR, Kang Y, Vukovich JM, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-DEC-2007) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall Snyder Hall Rm 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 4833)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4833
                     /organism="Cloning vector pEX18Ap-pheS"
                     /mol_type="other DNA"
                     /label=suicidal gene replacement vector
                     /note="suicidal gene replacement vector"
                     /db_xref="taxon:496656"
     CDS             complement(315..1328)
                     /codon_start=1
                     /gene="pheS"
                     /product="PheS"
                     /label=pheS
                     /note="phenylalanine t-RNA synthetase"
                     /protein_id="ABY73722.1"
                     /translation="MDLDQIVADAQQSFEGAADITTLENEKARFLGKSGALTELLKGLG
                     KLDPETRKTEGARINVAKQQVEAALNARRQALADALLNQRLAAEAIDVTLPGRGAGAGS
                     LHPVMRTWERVEQIFRSIGFDVADGPEIETDWYNFTALNSPENHPARSMQDTFYVDGKD
                     ADGRPLLLRTHTSPMQVRYARMNRPPIKVIAPGRTYRVDSDATHSPMFNQVEGLWIDEN
                     VSFADLKGAYTDFLKKFFERDDILVRFRPSYFPFTEPSAEIDMMFEHGKNAGKWLEISG
                     SGQVHPTVIRNMGLDPERYIGFGFGSGLERLTMLRYGVQDLRLFFENDLRFLRQFA"
     gene            complement(315..1328)
                     /gene="pheS"
                     /label=pheS
     regulatory      complement(1364..1405)
                     /regulatory_class="promoter"
                     /note="PS12"
     misc_feature    1423..1919
                     /label=contains origin of transfer
                     /note="contains origin of transfer"
     oriT            1441..1550
                     /note="incP origin of transfer"
     regulatory      1971..2296
                     /regulatory_class="terminator"
                     /note="transcriptional terminators T1T2"
     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="lacZ-alpha"
                     /product="beta-D-galactosidase alpha peptide"
                     /label=lacZ-alpha
                     /protein_id="ABY73723.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     gene            complement(2332..2616)
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     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"
     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."
     misc_feature    2830..3673
                     /label=region containing ColE1 origin of replication
                     /note="region containing ColE1 origin of replication"
     rep_origin      complement(3014..3602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3773..4633)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABY73724.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4634..4738)
                     /gene="bla"
                     /label=AmpR promoter