DF-UAS-spa-GCaMP6f
(Plasmid #67568)

• Purpose: Zebrafish 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: pDestTol2pA2 (modified)
• Backbone manufacturer: Drew Friedman (modification)
• Backbone size w/o insert (bp): 4500
• Total vector size (bp): 6000
• Modifications to backbone: a 10x UAS promoter and SV40 polyA
• Vector type: zebrafish transgenesis (see :A Spinal Opsin Controls Early Neural Activity and Drives a Behavioral Light Response)

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: spa-GCaMP6f
• Alt name: superfolder photoactivatable GCaMP6f
• Species: Synthetic
• Insert Size (bp): 1353
• Mutation: K65T, I83V, G87S, Y92D, V115H, K118V, S187R, Y196N, L221F, T222S, I250V, F256S, N262T
• Tag / Fusion Protein:
  • HIS-tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AscI (not destroyed)
• 3′ cloning site: MluI (not destroyed)
• 5′ sequencing primer: GFP reverse
• 3′ sequencing primer: SV40 reverse

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
  • National Institute of Genetics Limited Use Label License for Tol2 System
• Industry Terms:
  • Not Available to Industry