Rlucrarr3-Flash2
(Plasmid #74130)

• Purpose: FlAsH-BRET reporter to monitor conformational changes in Arrestin3. Insertion of FlAsH motif (CCPGCC) following amino acid residue 139 of Arrestin3.
• Depositing Lab: Louis Luttrell
• Publication: Lee et al Nature. 2016 Mar 31;531(7596):665-8. doi: 10.1038/nature17154. Epub 2016 Mar 23.

Backbone

• Vector backbone: pcDNA3.1-RLucC2
• Backbone manufacturer: gift from M. Bouvier (University of Montreal)
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Arrestin3
• Alt name: Arr3, beta-Arrestin2
• Species: R. norvegicus (rat)
• Mutation: Insertion of FlAsH motif (CCPGCC) following amino acid residue 139 (see depositor comment below).
• Tag / Fusion Protein:
  • Renilla luciferase (rLuc) (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (unknown if destroyed)
• 3′ cloning site: ApaI (unknown if destroyed)
• 5′ sequencing primer: CMV-forward
• 3′ sequencing primer: BGH-Rev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pcDNA3.1-rLuc-arrestin3 backbone was a gift from M. Bouvier (University of Montreal, Montreal Quebec, Canada)
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Renilla Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Note: during Addgene's quality control, it was found that the FlAsH motif was inserted after amino acid 139, not 140. This difference has NO affect on plasmid function.