|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||16414||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepBluescript SK+
- Backbone size w/o insert (bp) 3000
Vector typeMammalian Expression, Cre/Lox
Selectable markersHygromycin, Zeocin
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert namePGK-HygR & ZeoR
- Cloning method Restriction Enzyme
- 5′ cloning site BstXI (unknown if destroyed)
- 3′ cloning site KpnI (unknown if destroyed)
- 5′ sequencing primer mPGK-F
- 3′ sequencing primer T7 (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
Part of vector system to facilitate and simplify the use of rAAV for creating KOs in human cells. The other vectors for this system, pAAV-MCS, pAAV-RC, and pHelper, can be purchased from Stratagene.
The sequence information is the sequence between linkers A and B.
See author's map and article for more information. In the author's map, restriction sites for the generation of pNeDaKO fragments used for fusion PCR are shown. A and B refer to linkers A and B; yellow arrows: loxP sites; PGK: phosphoglycerate kinase eukaryotic promoter; Hyg: hygromycin resistance gene; EM 7: EM7 prokaryotic promoter; Zeo: zeomycin resistance gene; Amp: ampicillin resistance gene.
Note that though the Hyg-R gene has a point mutation, the Vogelstein lab has used this construct to for Hyg selection extensively.
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:pNeDaKO-Hyg was a gift from Bert Vogelstein (Addgene plasmid # 16414 ; http://n2t.net/addgene:16414 ; RRID:Addgene_16414)
For your References section:Facile methods for generating human somatic cell gene knockouts using recombinant adeno-associated viruses. Kohli M, Rago C, Lengauer C, Kinzler KW, Vogelstein B. Nucleic Acids Res. 2004 . 32(1):e3. 10.1093/nar/gnh009 PubMed 14704360
Map uploaded by the depositor.