Basic Vector Information
- Vector Name:
- pwFRT-pheS-gat
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4531 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Kang Y, Norris MH, Hoang TT.
pwFRT-pheS-gat 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-pheS-gat vector Sequence
LOCUS 40924_46538 4531 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pwFRT-pheS-gat, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4531)
AUTHORS Kang Y, Norris MH, Hoang TT.
TITLE Knock-out and pull-out recombineering protocols for natural
transformable Burkholderia thailandensis and Burkholderia
pseudomallei
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4531)
AUTHORS Kang Y, Norris MH, Hoang TT.
TITLE Direct Submission
JOURNAL Submitted (09-OCT-2009) Molecular Biosciences and Biological
Engineering, University of Hawaii at Manoa, 2538 McCarthy Mall,
Snyder Hall 207, Honolulu, HI 96822, USA
REFERENCE 3 (bases 1 to 4531)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4531)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-OCT-2009) Molecular Biosciences and Biological Engineering,
University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder Hall 207,
Honolulu, HI 96822, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4531
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..200
/label=AmpR promoter
CDS 201..1058
/label=AmpR
/note="beta-lactamase"
rep_origin 1232..1820
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2108..2129
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2144..2174
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2182..2198
/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 2206..2222
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2289..2356
/note="wild type FRT; Flp recognition target"
protein_bind complement(2306..2353)
/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 2393..2434
/note="PCS12; rpsL promoter from Burkholderia cenocepacia"
/regulatory_class="promoter"
RBS 2445..2467
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2478..3488
/gene="pheS"
/label=pheS
/note="Phenylalanine--tRNA ligase alpha subunit from
Burkholderia pseudomallei (strain 1710b). Accession#:
Q3JT08"
CDS 3527..3967
/codon_start=1
/gene="gat"
/product="glyphosate acetyl transferase"
/label=gat
/note="confers resistance to glyphosate"
/protein_id="ADO63837.1"
/translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
gene 3527..3967
/gene="gat"
/label=gat
protein_bind complement(4023..4070)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind complement(4137..4153)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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