pcDNA3-PM-InPAkt
(Plasmid #181915)

• Purpose: Indicator of Phosphoinositides using Akt; targeted to the plasma membrane.
• Depositing Lab: Jin Zhang
• Publication: Ananthanarayanan et al Proc Natl Acad Sci U S A. 2005 Oct 18;102(42):15081-6. doi: 10.1073/pnas.0502889102. Epub 2005 Oct 7.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5517
• Total vector size (bp): 7551
• Vector type: Mammalian Expression
• 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: pm-InPAkt
• Alt name: InPAkt-Kras
• Alt name: truncECFP-PH(Akt)-pseudoligand-Citrine-Kras
• Insert Size (bp): 2034
• Mutation: Contains the following mutations with respect to original Citrine: D21V; A206K; GGSGGT linker between L231 and G232
• Promoter: CMV
• Tags / Fusion Proteins:
  • 6xHIS - T7 tag (gene 10 leader) - Xpress (TM) tag (N terminal on backbone)
  • truncECFP (N terminal on insert)
  • Citrine (C terminal on insert)
  • C-terminal targeting sequence from Kras (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-InPAkt-Kras.gb

Terms and Licenses

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