pTriEx-PA-Rac1-C450A
(Plasmid #22025)

• Purpose: Expression of Rac1 fused with light-insensitive LOV2 (C450A mutation)
• Depositing Lab: Klaus Hahn
• Publication: Wu et al Nature. 2009 Sep 3. 461(7260):104-8.

Backbone

• Vector backbone: pTriEx
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 6000
• Vector type: Mammalian Expression, Bacterial Expression, Insect Expression

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-C450A
• Alt name: LOV(Leu404-Leu546 , C450A)-Rac1(Ile4)-Q61L/E91H/N92H
• Species: H. sapiens (human); Avena sativa (oat)
• Insert Size (bp): 1100
• Mutation: LOV domain contains light-insensitive mutation C450A.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)
• Promoter: CMV, p10
• Tag / Fusion Protein:
  • 6X His (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Bam HI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: pTriExUP

Resource Information

• Articles Citing this Plasmid:
  • 2 References

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.