Basic Vector Information
- Vector Name:
- pwFRT-GSr
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3507 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Norris MH, Kang Y, Lu D, Wilcox BA, Hoang TT.
pwFRT-GSr vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pwFRT-GSr vector Sequence
LOCUS 40924_46518 3507 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pwFRT-GSr, complete sequence.
ACCESSION .
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)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 3507)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2009"; volume: "75"; issue: "19";
pages: "6062-6075"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(14-OCT-2008) Molecular Biology and Bio-Engineering, University of
Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, Honolulu, HI 96822,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(07-APR-2009) Molecular Biology and Bio-Engineering, University of
Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, Honolulu, HI 96822,
USA"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT On Apr 7, 2009 this sequence version replaced FJ384986.1.
FEATURES Location/Qualifiers
source 1..3507
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 104..125
/label=CAP binding site
/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
/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
/label=PCS12
/note="PCS12"
/regulatory_class="promoter"
RBS 441..463
/label=RBS
/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
protein_bind complement(995..1042)
/label=FRT
/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
/label=AmpR promoter
CDS 1704..2561
/label=AmpR
/note="beta-lactamase"
rep_origin 2735..3323
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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