Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017163 | pSF3-Flag-CBir-FRB_Myc-NBir-FKBP | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pSF3-Flag-CBir-FRB_Myc-NBir-FKBP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8529 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Retrovirus
- Selection Marker:
- Hyg
- Promoter:
- minimal CMV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pSF3-Flag-CBir-FRB_Myc-NBir-FKBP vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pSF3-Flag-CBir-FRB_Myc-NBir-FKBP vector Sequence
LOCUS V017163 8529 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017163
VERSION V017163
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8529)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8529
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 387..979
/label="LTR"
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 1042..1399
/label="MMLV Psi"
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1473..1880
/label="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
polyA_signal 2058..2192
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(2382..2705)
/label="FKBP"
/note="human FK506-binding protein FKBP12"
CDS 3278..3298
/note="Neuropeptide F from Helicoverpa zea. Accession#:
P85106"
/label="Neuropeptide F"
CDS complement(3534..3563)
/label="Myc"
/note="Myc (human c-Myc proto-oncogene) epitope tag"
protein_bind complement(3715..3985)
/label="tetracycline response element"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 4000..4038
/label="minimal CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 4150..4173
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 4441..4719
/label="FRB"
/note="FKBP-rapamycin binding domain of human FRAP"
protein_bind 4760..4807
/label="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind complement(5891..5907)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 6381..6485
/label="AmpR promoter"
CDS 6486..7343
/label="AmpR"
/note="beta-lactamase"
rep_origin 7517..8105
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 8393..8414
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8429..8459
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 8467..8483
/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 8491..8507
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"