Basic Vector Information
- Vector Name:
- p3xFLAG-CMV-8
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4763 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Sigma-Aldrich
- Copy Number:
- High copy number
- Promoter:
- CMV
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
p3xFLAG-CMV-8 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 ℃.
p3xFLAG-CMV-8 vector Sequence
LOCUS p3xFLAG-CMV-8. 4763 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector for expression and secretion of N-terminally 3xFLAG-tagged
proteins in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS p3xFLAG-CMV-8.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4763)
AUTHORS Sigma-Aldrich
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4763)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4763
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 141..157
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 318..697
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 698..901
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 932..976
/label=preprotrypsin leader
/note="leader sequence from mouse preprotrypsin"
CDS 977..1042
/label=3xFLAG
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
misc_feature 1040..1106
/label=MCS
/note="MCS"
/note="multiple cloning site"
polyA_signal 1104..1726
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
promoter 1755..2084
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter complement(2123..2141)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2155..2171)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2179..2195)
/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)."
promoter complement(2203..2233)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2248..2269)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2557..3145)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3319..4176)
/label=AmpR
/note="beta-lactamase"
promoter complement(4177..4281)
/label=AmpR promoter
rep_origin 4308..4763
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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