pAAV-Ef1a-Con/Fon-taCasp3-T2A-TEVp-WPRE3-BGHpA
(Plasmid #242456)

• Purpose: AAV plasmid for expressing taCaspase-3 and T2A linked TEVp for caspase mediated cell ablation, and an intersectional strategy for Cre-off and Flp-on expression. See comments for more detail.
• Depositing Lab: Andrew Steele
• Publication: Coff/Fon Caspase3 (unpublished)

Backbone

• Vector backbone: pAAV[Exp]:WPRE
• Backbone manufacturer: Vector Builder
• Backbone size w/o insert (bp): 4986
• Total vector size (bp): 6603
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Caspase 3
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 1617
• Entrez Gene: CASP3 (a.k.a. CPP32, CPP32B, SCA-1)
• Promoter: Ef1a

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: Ef1a

Resource Information

• Supplemental Documents:
  • VB250604-1113wje.gb
• A portion of this plasmid was derived from a plasmid made by: Vector Builder

Terms and Licenses

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

Depositor Comments

This AAV construct expresses taCaspase-3 and T2A linked TEVp using the design of Dr. Nirao Shah for caspase mediated cell ablation (Yang et al. 2013 Cell. https://doi.org/10.1016/j.cell.2013.04.017. PubMed 23663785) and an intersectional strategy for Cre-off and Flp-on expression.
Note that the Intersect strategy used here does not make use of "introns" as initially designed by the Deisseroth lab (Fenno et al. 2014 Nat Methods. https://doi.org/10.1038/nmeth.2996. PubMed 24908100). Rather it uses loxP sites in the same orientation to facilitate a Cre-mediated deletion for "off" and inverted FRT sites to facilitate a Flp-mediated inversion for "on".