pBabe puro HA PIK3CA
(Plasmid #12522)

• Purpose: Retroviral expression of HA tagged phosphoinositide 3-kinase
• Depositing Lab: Jean Zhao
• Publication: Zhao et al Proc Natl Acad Sci U S A. 2005 Dec 20. 102(51):18443-8.

Backbone

• Vector backbone: pBabe puro
• Backbone manufacturer: Available at Addgene (plasmid #1764)
• Backbone size w/o insert (bp): 5150
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: PIK3CA
• Alt name: phosphoinositide 3-kinase
• Alt name: PI3K
• Alt name: p110 alpha
• Species: H. sapiens (human)
• Insert Size (bp): 3277
• Entrez Gene: PIK3CA (a.k.a. CCM4, CLAPO, CLOVE, CWS5, HMH, MCAP, MCM, MCMTC, PI3K, PI3K-alpha, p110-alpha)
• Tag / Fusion Protein:
  • HA (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: pBABE 5'
• 3′ sequencing primer: pBABE-3

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest for 12522
• Articles Citing this Plasmid:
  • 19 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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.