pBABEpuro-ERBB2 vector (V000698)

COA reports Sequencing Results MSDS Download GenBank File(.gb)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V000698	pBABEpuro-ERBB2	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

Antibiotic Resistance:: Ampicillin

Length:: 8907 bp

Type:: Mammalian Expression, Retroviral

Replication origin:: ori

Selection Marker:: Puromycin

Copy Number:: High Copy

Promoter:: SV40

Cloning Method:: Gateway Cloning

5' Primer:: tttgtacaccctaagcctc

3' Primer:: gcatacttctgcctgctg

pBABEpuro-ERBB2 vector Map

Copy Sequence View 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 vector Sequence

LOCUS       V000698                 8907 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V000698
VERSION     V000698
KEYWORDS    pBABEpuro-ERBB2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8907)
  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 8907)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8907)
  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"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    542..741
                     /label="MMLV Psi"
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             942..1358
                     /label="gag (truncated)"
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     protein_bind    1386..1410
                     /gene="mutant version of attB"
                     /label="attB1"
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1410..5174
                     /gene="ERBB2"
                     /label="Receptor tyrosine-protein kinase erbB-2"
                     /note="Receptor tyrosine-protein kinase erbB-2 from Homo
                     sapiens. Accession#: P04626"
     protein_bind    complement(5179..5199)
                     /label="attB2"
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     promoter        5242..5571
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5581..6177
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             6279..6870
                     /label="LTR"
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     primer_bind     complement(7000..7017)
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7171..7759)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7922..8779)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8780..8884)
                     /label="AmpR promoter"