Addgene - pBabe puro HA PIK3CA Plasmid Data

Plasmid 12522: pBabe puro HA PIK3CA

Gene/insert name:
PIK3CA
Alt name:
phosphoinositide 3-kinase
Alt name:
PI3K
Alt name:
p110 alpha
Insert size:
3277
Species:
H. sapiens (human)
Entrez Gene:
PIK3CA (CLOVE, CWS5, MCAP, MCM, MCMTC, PI3K, p110-alpha)
Fusion protein or tag:
HA
Terminal:
N terminal on insert
Vector backbone:
pBabe puro
(Search Vector Database)
Backbone manufacturer:
Available at Addgene (plasmid #1764)
Vector type:
Mammalian Expression, Retroviral
Backbone size w/o insert (bp):
5150
Cloning site 5':
BamHI
Site destroyed during cloning:
No
Cloning site 3':
SalI
Site destroyed during cloning:
No
5' sequencing primer:
pBABE 5' List of Sequencing Primers
3' sequencing primer:
pBABE-3
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Selectable markers:
Puromycin
Sequence: View sequences (2)
Supplemental document:
Notes from Addgene (1)
Principal Investigator:
Jean Zhao
Terms and Licenses:
MTA

Comments:

To construct a hemagglutinin (HA)-tagged allele of p110 alpha, a cDNA clone of human PIK3CA (obtained from the FLEXGene Repository of the Harvard Institute of Proteomics [HIP]) was subcloned into pBabepuro by PCR with primers containing HA-epitope coding sequence with subsequent restriction and ligation.

Please note that the original cDNA from HIP differed from the sequence posted on NCBI. This cDNA has an A->G alteration at the 424th nucleotide, which changes the amino acid from Ile -> Val. It could be a SNP. No functional changes have been observed so far in signaling assays.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: The oncogenic properties of mutant p110alpha and p110beta phosphatidylinositol 3-kinases in human mammary epithelial cells. Zhao et al (Proc Natl Acad Sci U S A. 2005 Dec 20. 102(51):18443-8. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 12522" in your Materials and Methods section.