pAAV-2Aneo v2
(Plasmid #80033)

• Purpose: (Empty Backbone) Used to construct AAV-based targeting vectors that carry an IVS-2Aneo cassette for promoter-trapping
• Depositing Lab: Hiroyuki Konishi
• Publication: Karnan et al Nucleic Acids Res. 2015 Dec 10. pii: gkv1338.

Backbone

• Vector backbone: pAAV-MCS
• Backbone manufacturer: Stratagene
• Backbone size (bp): 2892
• Modifications to backbone: a NotI-NotI fragment containing pAAV-MCS backbone was used for the cloning of pAAV-2Aneo v2.
• Vector type: AAV, Cre/Lox ; a platform vector to construct targeting vectors by inserting 5' and 3' arms
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: SK primer
• 3′ sequencing primer: KS primer

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Upon construction of a targeting vector with pAAV-2Aneo v2, the “frame adjuster” should be cleaved with BspEI or BsrGI and then self-religated, or left untreated, so that 2Aneo is translated in frame with the 5' portion of the targeted endogenous gene. This truncation of the frame adjuster should be done prior to the incorporation of 5' and 3' arms into pAAV-2Aneo v2, if arms carry restriction enzyme sites cleaved during the truncation of the frame adjuster (i.e., BspEI or BsrGI sites). This plasmid and pAAV-2Aneo differ only by the sequences of the frame adjusters.

A following article describes how to use pAAV-2Aneo v2 to construct AAV-based targeting vectors: Karnan et al. Bio-Protoc, 6(24): e2058, 2016. (http://www.bio-protocol.org/e2058)