pAAV-2Aneo
(Plasmid #80032)

• 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.
• 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

Terms and Licenses

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

Depositor Comments

Upon construction of a targeting vector with pAAV-2Aneo, the “frame adjuster” should be first cleaved with BspEI, MluI, or BsrGI and then self-religated 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, if arms carry restriction enzyme sites cleaved during the truncation of the frame adjuster (i.e., BspEI, MluI, or BsrGI sites). This plasmid and pAAV-2Aneo v2 differ only by the sequences of the frame adjusters.

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