Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000034 | pBABEpuro-ERBB2 A775_G776insYVMA | 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:
- pBABEpuro-ERBB2 A775_G776insYVMA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8919 bp
- Type:
- Mammalian Expression, Retroviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Cloning Method:
- Gateway Cloning
- 5' Primer:
- TTTGTACACCCTAAGCCTC
- 3' Primer:
- GCATACTTCTGCCTGCTG
pBABEpuro-ERBB2 A775_G776insYVMA vector 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.
pBABEpuro-ERBB2 A775_G776insYVMA vector Sequence
LOCUS 40924_5444 8919 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8919)
AUTHORS Greulich H, Kaplan B, Mertins P, Chen TH, Tanaka KE, Yun CH, Zhang
X, Lee SH, Cho J, Ambrogio L, Liao R, Imielinski M, Banerji S,
Berger AH, Lawrence MS, Zhang J, Pho NH, Walker SR, Winckler W, Getz
G, Frank D, Hahn WC, Eck MJ, Mani DR, Jaffe JD, Carr SA, Wong KK,
Meyerson M
TITLE Functional analysis of receptor tyrosine kinase mutations in lung
cancer identifies oncogenic extracellular domain mutations of ERBB2.
JOURNAL Proc Natl Acad Sci U S A. 2012 Sep 4;109(36):14476-81. Epub 2012 Aug
20.
PUBMED 22908275
REFERENCE 2 (bases 1 to 8919)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8919)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A."; date: "2012-09-4"; volume: "109(36)"; pages:
"14476-81. Epub 2012 Aug 20"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8919
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(933..1790)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1791..1895)
/label=AmpR promoter
LTR 1926..2395
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 2460..2659
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 2860..3276
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
protein_bind 3304..3328
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(7109..7129)
/label=attB2
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 7172..7501
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 7511..8107
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
LTR 8209..8800
/label=LTR
/note="long terminal repeat from Moloney murine leukemia
virus"