Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012084 | EGFR L858R | 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:
- EGFR L858R
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8725 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- pBABE5'
EGFR L858R 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.
EGFR L858R vector Sequence
LOCUS EGFR_L858R. 8725 bp DNA circular SYN 13-MAY-2021
DEFINITION Retroviral construct for expressing human EGFR with L858R mutation
in human cells.
ACCESSION .
VERSION .
KEYWORDS EGFR L858R
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8725)
AUTHORS Greulich H, Chen TH, Feng W, Janne PA, Alvarez JV, Zappaterra M,
Bulmer SE, Frank DA, Hahn WC, Sellers WR, Meyerson M
TITLE Oncogenic Transformation by Inhibitor-Sensitive and -Resistant EGFR
Mutants.
JOURNAL PLoS Med. 2005 Oct 4. 2(11):e313.
PUBMED 16187797
REFERENCE 2 (bases 1 to 8725)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8725)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Med.
2005 Oct 4. 2(11):e313."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8725
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..3630
/label=EGFR
/note="precursor of human epidermal growth factor receptor"
promoter 3675..4004
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4014..4610
/label=PuroR
/note="puromycin N-acetyltransferase"
LTR 4712..5303
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5433..5450)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5604..6192)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6355..7212)
/label=AmpR
/note="beta-lactamase"
promoter complement(7213..7317)
/label=AmpR promoter
LTR 7348..7817
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 7882..8081
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 8282..8698
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"