Basic Vector Information
- Vector Name:
- prCol2(P)-FRT
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4899 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Ito K, Shinomura T.
- Promoter:
- SP6
prCol2(P)-FRT 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 ℃.
prCol2(P)-FRT vector Sequence
LOCUS 40924_36408 4899 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector prCol2(P)-FRT DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4899)
AUTHORS Ito K, Shinomura T.
TITLE Development and application of a new Silent reporter system to
quantitate the activity of enhancer elements in the type II Collagen
Gene
JOURNAL Gene (2016) In press
PUBMED 26992640
REFERENCE 2 (bases 1 to 4899)
AUTHORS Shinomura T.
TITLE Direct Submission
JOURNAL Submitted (06-OCT-2015) Contact:Tamayuki Shinomura Tokyo Medical and
Dental University, Tissue Regeneration; 1-5-45 Yushima, Bunkyo-ku,
Tokyo 113-8549, Japan
REFERENCE 3 (bases 1 to 4899)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4899)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene (2016)
In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-OCT-2015) Contact:Tamayuki Shinomura Tokyo Medical and Dental
University, Tissue Regeneration; 1-5-45 Yushima, Bunkyo-ku, Tokyo
113-8549, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4899
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 14..32
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
gene 78..1727
/gene="Col2a1"
/label=Col2a1
regulatory 78..1066
/gene="Col2a1"
/label=rat type II collagen
/note="rat type II collagen"
/regulatory_class="promoter"
exon 1067..1307
/gene="Col2a1"
/number=1
/note="rat type II collagen"
intron 1308..1351
/gene="Col2a1"
/number=1
/note="rat type II collagen partial intron"
intron 1556..1710
/gene="Col2a1"
/number=2
/note="rat type II collagen partial intron"
exon 1711..1727
/gene="Col2a1"
/number=3
/note="rat type II collagen partial exon"
protein_bind 1736..1783
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter complement(1798..1816)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1823..1839)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(1980..2435)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2769..2873
/label=AmpR promoter
CDS 2874..3731
/label=AmpR
/note="beta-lactamase"
rep_origin 3905..4493
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 4781..4802
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4817..4847
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4855..4871
/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 4879..4895
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
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