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14-3-3 KO vector
(Plasmid #16551)

Full plasmid sequence is not available for this item.

Ordering

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

  • Vector backbone
    pBluescript
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 3000
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    14-3-3 knockout
  • Alt name
    14-3-3 sigma
  • Species
    H. sapiens (human)
  • Entrez Gene
    SFN (a.k.a. YWHAS)

Cloning Information

  • 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)

Resource Information

  • Terms and Licenses

Depositor Comments

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.

How to cite this plasmid ( Back to top)

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