pcDNA3-PKCsub(T/A)-YPet-NES
(Plasmid #138208)

• Purpose: Encodes negative-control C-terminal (substrate) fragment of FRET-based bimolecular PKC activity reporter (bimCKAR); cytosol targeted. Use in conjunction with pcDNA3-Cerulean-FHA1-NES
• Depositing Lab: Jin Zhang
• Publication: Herbst et al J Am Chem Soc. 2011 Apr 20;133(15):5676-9. doi: 10.1021/ja1117396. Epub 2011 Mar 25.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5472
• Total vector size (bp): 6288
• 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: PKCsub(T/A)-YPet-NES
• Species: Synthetic
• Insert Size (bp): 816
• Mutation: Target Thr residue in substrate domain mutated to Ala.
• Promoter: CMV
• Tags / Fusion Proteins:
  • YPet (C terminal on insert)
  • Nuclear export signal (NES) (C terminal on insert)
  • 6xHIS - T7 tag (gene 10 leader) - Xpress (TM) tag (N terminal on backbone)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pcDNA3-PKCsub(TA)-YPet-NES.gb

Terms and Licenses

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