Bcr GAP domain R1090A/pGEX 3X
(Plasmid #36406)

• Depositing Lab: Nora Heisterkamp
• Publication: Cho et al Mol Cell Biol. 2007 Feb;27(3):899-911. Epub 2006 Nov 20.

Backbone

• Vector backbone: pGEX 3X
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Rosetta2
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Bcr GAP domain encoding residues 1004-1271 with R1090A point mutation
• Species: H. sapiens (human)
• Insert Size (bp): 1300
• Entrez Gene: BCR (a.k.a. ALL, BCR1, CML, D22S11, D22S662, PHL)
• Tag / Fusion Protein:
  • GST (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pGEX-3X
• 3′ sequencing primer: pGEX-3X

Terms and Licenses

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

Depositor Comments

We constructed this in a 3-way ligation. The 5' end was a 0.16 kb BamHI-BstEII fragment from plasmid Bcr GAP domain in pGEX-3X, which provided the 5' BamHI site from pGEX-3X. The 3' end was a 1.2 kb BstEII-EcoRI fragment carrying the R1090A mutation. This was cloned into pGEX-3X digested with BamHI-EcoRI. The construct encodes the GAP-dead domain of Bcr which is inactive against GTP-bound Rac. It includes the natural stop codon and 3' untranslated sequences of the cDNA. Constructed by Nora Heisterkamp. The Rosetta2 strain may improve yield of the fusion protein.