Ratiometric FPX sensor for ERK kinase activity
(Plasmid #60974)

• Purpose: For imaging of ERK kinase activity using a single polypeptide RA-WW domain-ERK substrate-B protein and free GA-NES.
• Depositing Lab: Robert Campbell
• Publication: Ding et al Nat Methods. 2015 Jan 26. doi: 10.1038/nmeth.3261.

Backbone

• Vector backbone: pcDNA3.0
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 7493
• Modifications to backbone: Multi cloning sites has been revised
• 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): DH10B
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ddRFP A-WW domain-ERK substrate-ddFP B
• Alt name: RA-WW domain-ERK substrate-B
• Species: Synthetic; synthetic construct
• Insert Size (bp): 1993
• GenBank ID: KM979350
• Promoter: CMV
• Tags / Fusion Proteins:
  • WW domain is after ddRFP-A
  • ERK substrate is before ddFP copy B
  • ddFP copy B is after ERK substrate

Cloning Information

• 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

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry