pGEX SH3(2)-4R
(Plasmid #112090)

• Purpose: Bacterial expression plasmid containing a GST tag with 4 repeats of the second SH3 domain from human NCK1.
• Depositing Lab: Michael Rosen
• Publication: Li et al Nature. 2012 Mar 7;483(7389):336-40. doi: 10.1038/nature10879.

Backbone

• Vector backbone: pGEX2T
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 6000
• Modifications to backbone: A Tev protease site has been added between the thrombin site and the multiple cloning site. The multiple cloning has been modified to begin with a NdeI (CATATG) site, and end with a BamHI (GGATCC) site.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: SH3(2)-4R
• Alt name: NCK adaptor protein 1, isoform 2, second SH3 repeat x 4
• Species: H. sapiens (human)
• Insert Size (bp): 997
• GenBank ID: NP_001177725.1
• Entrez Gene: NCK1 (a.k.a. NCK, NCKalpha, nck-1)
• Promoter: Lac
• Tag / Fusion Protein:
  • GST (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: pGEX-5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3'
• 3′ sequencing primer: pGEX-3’: 5'-CCGGGAGCTGCATGTGTCAGAGG-3'

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

There are two NdeI sites - NdeI was used to make the additional repeats.