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PurposeRetroviral expression of HA tagged phosphoinositide 3-kinase H1047R (Tumor-derived allele)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 12524 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepBabe puro
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Backbone manufacturerAvailable at Addgene (plasmid #1764)
- Backbone size w/o insert (bp) 5150
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Vector typeMammalian Expression, Retroviral
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Selectable markersPuromycin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namePIK3CA H1047R
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Alt namephosphoinositide 3-kinase
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Alt namePI3K
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Alt namep110 alpha
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SpeciesH. sapiens (human)
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Insert Size (bp)3277
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MutationH1047R. Tumor-derived allele.
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Entrez GenePIK3CA (a.k.a. CCM4, CLAPO, CLOVE, CWS5, HMH, MCAP, MCM, MCMTC, PI3K, PI3K-alpha, p110-alpha)
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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' (Common Sequencing Primers)
Resource Information
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Mutation generated using Strategene Quickchange site-directed mutagenesis kit. See article for primer information.
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.
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.
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For your Materials & Methods section:
pBabe puro HA PIK3CA H1047R was a gift from Jean Zhao (Addgene plasmid # 12524 ; http://n2t.net/addgene:12524 ; RRID:Addgene_12524) -
For your References section:
The oncogenic properties of mutant p110alpha and p110beta phosphatidylinositol 3-kinases in human mammary epithelial cells. Zhao JJ, Liu Z, Wang L, Shin E, Loda MF, Roberts TM. Proc Natl Acad Sci U S A. 2005 Dec 20. 102(51):18443-8. 10.1073/pnas.0508988102 PubMed 16339315