14-3-3 KO vector
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||16551||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 3000
Vector typeBacterial Expression
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert name14-3-3 knockout
Alt name14-3-3 sigma
SpeciesH. sapiens (human)
Entrez GeneSFN (a.k.a. YWHAS)
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer T3 (Common Sequencing Primers)
Terms and Licenses
A BAC clone was digested with BamHI, and two fragments, one 4.7 kilobases (kb) and the second 3.8 kb, were used to construct the 5' and 3' arms of the targeting vector, respectively. The 4.7-kb subclone contained the region immediately 5' of the initiating codon of the 14-3-3sigma coding region. The 3.8-kb subclone contained a region beginning 900 bp distal to the initiating codon. A synthetic EcoRV site plus the sequence 5'-CGTGGAGAGGGACT GGCAG-3', derived from a region of the 14-3-3sigma locus deleted by the targeting construct, were ligated to the 5' end of a geneticin-resistance gene. This product was placed into the pBluescript plasmid. To facilitate knockout of both alleles, loxP sites surrounding the geneticin-resistance gene were incorporated into the vector.
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:14-3-3 KO vector was a gift from Bert Vogelstein (Addgene plasmid # 16551 ; http://n2t.net/addgene:16551 ; RRID:Addgene_16551)
For your References section:14-3-3Sigma is required to prevent mitotic catastrophe after DNA damage. Chan TA, Hermeking H, Lengauer C, Kinzler KW, Vogelstein B. Nature. 1999 Oct 7. 401(6753):616-20. 10.1038/44188 PubMed 10524633
Map uploaded by the depositor.