LOCUS       Exported                4565 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pSHAFT-GFP, complete sequence.
ACCESSION   KX485331
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4565)
  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 4565)
  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 4565)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4565
                     /organism="Suicide vector pSHAFT-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1918679"
     CDS             complement(10..729)
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /protein_id="APD15320.1"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYN"
     gene            complement(10..729)
                     /gene="gfp"
                     /label=gfp
     RBS             737..759
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    898..916
                     /label=O end
                     /note="O end"
     protein_bind    928..944
                     /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)."
     rep_origin      979..1367
                     /note="oriR6K; gamma replication origin from E. coli 
                     plasmid R6K"
     CDS             complement(1985..2356)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /protein_id="APD15321.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(1985..2356)
                     /gene="traJ"
                     /label=traJ
     oriT            2389..2498
                     /label=incP origin of transfer
                     /note="incP origin of transfer"
     CDS             complement(3576..4436)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers resistance to ampicillin"
                     /protein_id="APD15322.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3576..4436)
                     /gene="bla"
                     /label=bla
     regulatory      complement(4437..4541)
                     /regulatory_class="promoter"
                     /note="bla promoter"
     promoter        complement(4437..4541)
                     /gene="bla"
                     /label=AmpR promoter