LOCUS       Exported                3507 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pwFRT-GSr, complete sequence.
ACCESSION   FJ384986
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3507)
  AUTHORS   Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
  TITLE     Glyphosate resistance as a novel select-agent-compliant, 
            non-antibiotic-selectable marker in chromosomal mutagenesis of the 
            essential genes asd and dapB of Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 75 (19), 6062-6075 (2009)
  PUBMED    19648360
REFERENCE   2  (bases 1 to 3507)
  AUTHORS   Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Molecular Biology and Bio-Engineering, 
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, 
            Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 3507)
  AUTHORS   Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-APR-2009) Molecular Biology and Bio-Engineering, 
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, 
            Honolulu, HI 96822, USA
REFERENCE   4  (bases 1 to 3507)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Apr 7, 2009 this sequence version replaced FJ384986.1.
FEATURES             Location/Qualifiers
     source          1..3507
                     /organism="Cloning vector pwFRT-GSr"
                     /mol_type="other DNA"
                     /db_xref="taxon:578228"
     protein_bind    104..125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        140..170
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    178..194
                     /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)."
     primer_bind     202..218
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    285..358
                     /label=wild type Flp recognition target
                     /note="wild type Flp recognition target"
     protein_bind    complement(302..349)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      389..430
                     /regulatory_class="promoter"
                     /note="PCS12"
     RBS             441..463
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             499..939
                     /codon_start=1
                     /gene="gat"
                     /product="Gat"
                     /label=gat
                     /note="glyphosate acetyl transferase; confers resistance to
                     glyphosate"
                     /protein_id="ACJ70055.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            499..939
                     /gene="gat"
                     /label=gat
     misc_feature    978..1051
                     /label=wild type Flp recognition target
                     /note="wild type Flp recognition target"
     protein_bind    complement(995..1042)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(1109..1125)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1599..1703
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1704..2564
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="ACJ70056.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1704..2564
                     /gene="bla"
                     /label=bla
     rep_origin      2735..3323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2771..3387
                     /label=ColE1 origin of replication
                     /note="ColE1 origin of replication"