Ratiometric FPX sensor for ERK kinase activity
PurposeFor imaging of ERK kinase activity using a single polypeptide RA-WW domain-ERK substrate-B protein and free GA-NES.
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||60974||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) 5500
- Total vector size (bp) 7493
Modifications to backboneMulti cloning sites has been revised
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert nameddRFP A-WW domain-ERK substrate-ddFP B
Alt nameRA-WW domain-ERK substrate-B
SpeciesSynthetic; synthetic construct
Insert Size (bp)1993
- Promoter CMV
/ Fusion Proteins
- WW domain is after ddRFP-A
- ERK substrate is before ddFP copy B
- ddFP copy B is after ERK substrate
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer T7 Promoter forward primer
- 3′ sequencing primer BGH Reverse primers (Common Sequencing Primers)
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:Ratiometric FPX sensor for ERK kinase activity was a gift from Robert Campbell (Addgene plasmid # 60974)
For your References section:Ratiometric biosensors based on dimerization-dependent fluorescent protein exchange. Ding Y, Li J, Enterina JR, Shen Y, Zhang I, Tewson PH, Mo GC, Zhang J, Quinn AM, Hughes TE, Maysinger D, Alford SC, Zhang Y, Campbell RE. Nat Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261. 10.1038/nmeth.3261 PubMed 25622108