This website uses cookies to ensure you get the best experience. By continuing to use this site, you agree to the use of cookies.

Please note: Your browser does not support the features used on Addgene's website. You may not be able to create an account or request plasmids through this website until you upgrade your browser. Learn more

Please note: Your browser does not fully support some of the features used on Addgene's website. If you run into any problems registering, depositing, or ordering please contact us at [email protected] Learn more

pLNCX Myr His p110 KD
(Plasmid #10837)

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 10837 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

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
  • 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
  • (Common Sequencing Primers)

Resource Information

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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pLNCX Myr His p110 KD was a gift from Thomas Roberts (Addgene plasmid # 10837)
  • For your References section:

    Constitutive cellular expression of PI 3-kinase is distinct from transient expression. Auger KR, Wang J, Narsimhan RP, Holcombe T, Roberts TM. Biochem Biophys Res Commun. 2000 Jun 16. 272(3):822-9. 10.1006/bbrc.2000.2806 PubMed 10860837