pECFP-IETD2x-Venus
(Plasmid #24536)

• Depositing Lab: Peter Sorger
• Publication: Albeck et al PLoS Biol. 2008 Dec 2. 6(12):2831-52.

Backbone

• Vector backbone: pECFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4731
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ECFP-IETD2x-Venus
• Alt name: Caspase-8/10 FRET reporter
• Alt name: Initiator caspase reporter protein (IC-RP)
• Species: Synthetic
• Insert Size (bp): 804
• Promoter: CMV
• Tags / Fusion Proteins:
  • ECFP
  • Venus

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BspEI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40pA-R

Terms and Licenses

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

Depositor Comments

Initiator caspase reporter protein (IC-RP) carries tandem copies of IETD in its linker, a sequence that is efficiently cleaved by caspase-8, but poorly by caspases-3 and -7. IETD constitutes the site in procaspase-3 for initiator caspase cleavage, and IC-RP cleavage is therefore a good readout of procaspase-3 activation.

IC-RP was constructed by ligating Venus(YFP) between BamHI and EcoRI sites of pECFP-C1 and ligating linkers encoding cleavage sequences as BspEI-BamHI fragments between ECFP and Venus. Multiple serine and glycine residues flanking the cleavage sequences were added to increase linker flexibility and substrate accessibility. See https://www.ncbi.nlm.nih.gov/pubmed/18406323 for more information