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pwFRT-GSr vector (V002141#)

Basic Vector Information

Basic Vector Information
Vector Name pwFRT-GSr Antibiotic Resistance Ampicillin
Length 3507 bp Type Cloning vector
Source Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.

pwFRT-GSr vector Vector Map

CAP binding site lac promoter lac operator M13 rev wild type Flp recognition target PCS12 RBS gat wild type Flp recognition target M13 fwd AmpR promoter bla ori

pwFRT-GSr vector Sequence

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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"

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