pTriEx-mCherry-PA-Rac1-C450A
(Plasmid #22028)

• Purpose: Expression of Rac1 fused with light-insensitive LOV2 (C450A mutation)
• Depositing Lab: Klaus Hahn
• Depositing Organization: University of North Carolina at Chapel Hill
  Located in United States of America
• Publication: Wu et al Nature. 2009 Sep 3. 461(7260):104-8.
• Citations: 5 Articles

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: LOV2-Rac1
• Species: H. sapiens (human); Avena sativa (oat)
• Insert Size (bp): 1100
• Mutation: LOV(Leu404-Leu546, C450A). Rac1 starts at Ile4 and contains mutations Q61L, E91H and N92H.
• GenBank ID: AAA36544
• Entrez Gene: RAC1 (a.k.a. MIG5, MRD48, Rac-1, TC-25, p21-Rac1)
• Promoter: CMV, p10
• Tags / Fusion Proteins:
  • 6xHis (N terminal on backbone)
  • mCherry (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

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.

DNA (Catalog # 22028-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 22028-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.