Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006650 | pCMV Flag WT-PTEN | 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:
- pCMV Flag WT-PTEN
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5539 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T3
- 3' Primer:
- T7
pCMV Flag WT-PTEN 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.
pCMV Flag WT-PTEN vector Sequence
LOCUS V006650 5539 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006650
VERSION V006650
KEYWORDS pCMV Flag WT-PTEN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 5539)
AUTHORS Mosessian S, Avliyakulov NK, Mulholland DJ, Boontheung P, Loo JA, Wu
H
TITLE Analysis of PTEN complex assembly and identification of hnRNP C as a
component of the PTEN-associated complex.
JOURNAL J Biol Chem. 2009 Sep 9. ():.
PUBMED 19740742
REFERENCE 2 (bases 1 to 5539)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5539)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2009 Sep 9. ():."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5539
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 68..371
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 621..639
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 674..683
/label="Kozak sequence"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 683..706
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 722..1930
/gene="PTEN"
/label="Phosphatidylinositol 3,4,5-trisphosphate
3-phosphatase and dual-specificity protein phosphatase
PTEN"
/note="Phosphatidylinositol 3,4,5-trisphosphate
3-phosphatase and dual-specificity protein phosphatase PTEN
from Canis lupus familiaris. Accession#: P60483"
primer_bind complement(1969..1985)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2042..2060)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2334..2455
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2462..2917)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2944..3046
/label="AmpR promoter"
promoter 3050..3407
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3442..4233
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
primer_bind complement(4424..4443)
/label="TK-pA-R"
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4468..4515
/label="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4844..5432
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"