Purpose(Empty Backbone) Used to construct AAV-based targeting vectors that carry an IVS-2Aneo cassette for promoter-trapping
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||80033||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size (bp) 2892
Modifications to backbonea NotI-NotI fragment containing pAAV-MCS backbone was used for the cloning of pAAV-2Aneo v2.
Vector typeAAV, Cre/Lox ; a platform vector to construct targeting vectors by inserting 5' and 3' arms
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Growth Strain(s)NEB Stable
Copy numberHigh Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer SK primer
- 3′ sequencing primer KS primer (Common Sequencing Primers)
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)
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:pAAV-2Aneo v2 was a gift from Hiroyuki Konishi (Addgene plasmid # 80033 ; http://n2t.net/addgene:80033 ; RRID:Addgene_80033)
For your References section:Improved methods of AAV-mediated gene targeting for human cell lines using ribosome-skipping 2A peptide. Karnan S, Ota A, Konishi Y, Wahiduzzaman M, Hosokawa Y, Konishi H. Nucleic Acids Res. 2015 Dec 10. pii: gkv1338. 10.1093/nar/gkv1338 PubMed 26657635