pBabe-TetCMV-puro-mVenus-PA-Rac1
(Plasmid #22008)

• Purpose: Mammalian expression of photoactivatable Rac1
• Depositing Lab: Klaus Hahn
• Publication: Wu et al Nature. 2009 Sep 3. 461(7260):104-8.

Backbone

• Vector backbone: pBabe-TetCMV-puro
• Backbone size w/o insert (bp): 6000
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: PA-Rac1
• Alt name: LOV2-Rac1
• Species: H. sapiens (human); Avena sativa (oat)
• Insert Size (bp): 1800
• Mutation: LOV(Leu404-Leu546), Rac1 starts at I4 and contains mutations Q61L, E91H and N92H.
• Entrez Gene: RAC1 (a.k.a. MIG5, MRD48, Rac-1, TC-25, p21-Rac1)
• Tag / Fusion Protein:
  • mVenus (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: pBabe-TetCMV Forward

Terms and Licenses

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

Depositor Comments

Protocols for production and use of Hahn lab biosensors and photoactivatable proteins can be found on their web page, at the URL below. If you have suggestions for the protocols or have any questions, please feel free to contact the Hahn lab. Good luck with your experiments!

Hahn lab biosensor protocol: http://www.hahnlab.com/tools/index.html

Please note, Addgene has not confirmed all the mutations associated with this construct. We suggest sequence verifying this plasmid upon receipt.