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pECFP-DEVDR-Venus
(Plasmid #24537)

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Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 24537 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

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-DEVDR-Venus
  • Alt name
    Caspase-3 FRET reporter
  • Alt name
    effector caspase reporter protein (EC-RP)
  • Species
    Synthetic
  • Insert Size (bp)
    792
  • 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
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Effector caspase reporter protein (EC-RP) used to monitor caspase-3 activity (and caspase-7 activity to a lesser extent). EC-RP construct is composed of a FRET donor-acceptor pair ECFP and Venus connected via a flexible linker (SGLRSSGDEVDRVYGSGS) that contains the caspase cleavage sequence DEVDR. When the linker is cleaved, energy transfer is lost and the ECFP signal increases. The DEVDR linker in EC-RP is expected to have 20-fold greater selectivity for caspase-3 relative to caspase-8 than the DEVDG linker used in other caspase reporters.

EC-RP was constructed by ligating Venus (YFP) between BamHI and EcoRI sites in 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.

How to cite this plasmid ( Back to top)

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:

    pECFP-DEVDR-Venus was a gift from Peter Sorger (Addgene plasmid # 24537 ; http://n2t.net/addgene:24537 ; RRID:Addgene_24537)
  • For your References section:

    Modeling a snap-action, variable-delay switch controlling extrinsic cell death. Albeck JG, Burke JM, Spencer SL, Lauffenburger DA, Sorger PK. PLoS Biol. 2008 Dec 2. 6(12):2831-52. 10.1371/journal.pbio.0060299 PubMed 19053173