|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||14860||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5400
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameC kinase activity reporter
/ Fusion Proteins
- CFP (N terminal on insert)
- YFP (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site XbaI (not destroyed)
- 5′ sequencing primer T7 (Common Sequencing Primers)
Terms and Licenses
Articles Citing this Plasmid
PKC substrate sequence flanked by flexible linker sequence: GGSGG RFRRFQTLKIKAKA GGSGG. A206K mutation present in both CFP (mCFP) and citrine (mYFP) to reduce the intrinsic homoaffinity of all GFPs and preclude intermolecular FRET by CFP-YFP dimerization. CFP was amplified by PCR from a plasmid template to encode a HindIII restriction site followed by a consensus initiation site for translation (CGCCACC) before the initiating ATG of CFP, and a KpnI restriction site at the 3' end instead of a terminating codon. FHA2, a gift from Michael Yaffe (MIT, Cambridge, MA) was amplified by PCR to include KpnI and BamHI restriction sites at the 5' and 3' ends, respectively. Citrine was amplified by PCR to include a 5' BamHI followed by the PKC substrate sequence and a 3' XbaI following the terminating codon.
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:CKAR was a gift from Alexandra Newton (Addgene plasmid # 14860 ; http://n2t.net/addgene:14860 ; RRID:Addgene_14860)
For your References section:A genetically encoded fluorescent reporter reveals oscillatory phosphorylation by protein kinase C. Violin JD, Zhang J, Tsien RY, Newton AC. J Cell Biol. 2003 Jun 9. 161(5):899-909. 10.1083/jcb.200302125 PubMed 12782683