pRSET-triple mutation-pa-GCaMP6f
(Plasmid #67570)

• Purpose: bacterial expression of a photoactivatable fluorescent calcium reporter
• Depositing Lab: Ehud Isacoff
• Publication: Berlin et al Nat Methods. 2015 Jul 13. doi: 10.1038/nmeth.3480.

Backbone

• Vector backbone: pRSET-B
• Backbone manufacturer: invitrogen
• Backbone size w/o insert (bp): 2897
• Total vector size (bp): 4250
• Modifications to backbone: introduction of BgLII mutation before start codon
• Vector type: Bacterial 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-GCaMP6f
• Alt name: photoactivatable GCaMP6f
• Species: Synthetic
• Insert Size (bp): 1353
• Mutation: V115H, L64F, T65S
• Promoter: T7
• Tag / Fusion Protein:
  • HIS-tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BgLII (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 reverse priming site

Resource Information

• A portion of this plasmid was derived from a plasmid made by: derived from Addgene clone #40755, published by Chen TW et al. article: ultrasensitive fluorescent proteins for imaging neuronal activity doi: 10.1038/nature12354

Terms and Licenses

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