pEGFP-Nck1 (M3)
(Plasmid #45896)

• Depositing Lab: Louise Larose
• Publication: Kebache et al Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5406-11.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Nck-1 M3 mutant
• Alt name: NCK1
• Alt name: NCK adaptor protein 1
• Species: H. sapiens (human)
• Mutation: M3 (W229R) in SH3 domain
• Entrez Gene: NCK1 (a.k.a. NCK, NCKalpha, nck-1)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40pA-R

Resource Information

• Supplemental Documents:
  • Nck1 schematic of mutations

Terms and Licenses

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

Depositor Comments

Wild type pEGFP-Nck1 (Addgene plasmid 45903) is described in JBC 26:26633-44 2006 (PMID 16835242). Mutations are described in the article linked to this plasmid (PMID 11959995) and then subcloned into the pEGFP-C1 vector using PCR amplification with primers containing XhoI and EcoRI sites.