pLNCX Myr His p110 KD
(Plasmid #10837)

• Depositing Lab: Thomas Roberts
• Publication: Auger et al Biochem Biophys Res Commun. 2000 Jun 16. 272(3):822-9.

Backbone

• Vector backbone: pLNCX
• Backbone manufacturer: BD Biosciences
• Backbone size w/o insert (bp): 6620
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Neomycin (select with G418)

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: p110 KD
• Alt name: PI3 Kinase
• Alt name: PI3K
• Alt name: PIK3CA
• Species: B. taurus (bovine)
• Mutation: D915S. Kinase Dead.
• GenBank ID: NM_174574
• Entrez Gene: PIK3CA
• Tags / Fusion Proteins:
  • Myr (N terminal on insert)
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: HindIII (destroyed during cloning)
• 5′ sequencing primer: CMV-F

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

A pair of overlapping oligomers were synthesized based on the c-src myristoylation signal peptide sequence. A Kozak sequence was engineered at the 5 of the c-src myristoylation signal sequence (MetGly-Ser-Ser-Lys-Ser-Lys-Pro-Lys-Asp-Pro-Ser-Gln-Arg-Arg-ArgArg-Ile-Arg-Thr). The fragment was ligated to the 5 end of bovine p110 sequence and cloned into a retroviral expression vector (pLNCX) by a three piece ligation reaction. A 6 histidine tag was incorporated on the 3 end of p110 by PCR. The kinase dead mutant has an additional BamHI site at D915S.