pGAD424-Rabex-5
(Plasmid #197043)

• Purpose: Expression of GAL4 transcriptional activation domain (AD)-Rabex-5 fusion protein in yeast (yeast two-hybrid assays)
• Depositing Lab: Juan Bonifacino
• Publication: Mattera et al J Biol Chem. 2006 Mar 10;281(10):6874-83. doi: 10.1074/jbc.M509939200. Epub 2006 Jan 5.

Backbone

• Vector backbone: pGAD424
• Backbone size w/o insert (bp): 6665
• Total vector size (bp): 8189
• Vector type: Yeast Expression
• Selectable markers: LEU2

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Rabex-5
• Species: B. taurus (bovine)
• Insert Size (bp): 1527
• Entrez Gene: RABGEF1 (a.k.a. RABEX5)
• Promoter: ADH1
• Tag / Fusion Protein:
  • GAL4 transcriptional activation domain (AD) fragment (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (destroyed during cloning)
• 3′ cloning site: BamH1 (destroyed during cloning)
• 5′ sequencing primer: GAL4 transcriptional activation domain forward
• 3′ sequencing primer: GAL4 transcriptional activation domain reverse

Terms and Licenses

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

Depositor Comments

NotI/SpeI fragment encoding Rabex-5 was polished wIth T4 DNA polymerse and subcloned into pGAD424 digested with BamH1 and also polished wIth T4 DNA polymerase. Insert contains an additional N-terminal GRSSP sequence preceding the Rabex-5 initiation codon.