Salsa6f
(Plasmid #140188)

• Purpose: Salsa6f is a fluorescent ratiometric genetically encoded calcium indicator, (tdTomato linked to GCaMP6f by a V5 epitope tag)
• Depositing Lab: Michael Cahalan
• Publication: Dong et al eLife. 2017 Dec 14;6. pii: 32417. doi: 10.7554/eLife.32417.

Backbone

• Vector backbone: pEGFP-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3923
• Total vector size (bp): 6698
• Modifications to backbone: The EGFP gene has been replaced with the ratiometric genetically encoded calcium indicator Salsa6f (tdTomato-V5-GCaMP6f)
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: Salsa6f
• Alt name: tdTomato-v5-GCaMP6f
• Species: Synthetic
• Insert Size (bp): 2775
• Promoter: CMV Immediate Early
• Tag / Fusion Protein:
  • tdTomato-v5-GCaMP6f

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: TGACGTCAATGGGAGTTTG
• 3′ sequencing primer: ACATTGATGAGTTTGGAC

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The fusion protein in this plasmid is the fluorescent genetically encoded calcium indicator Salsa6f, which is a fusion of the dimeric red fluorescent protein tdTomato, the epitope tag V5, and the green fluorescent genetically encoded calcium indicator GCaMP6f, in order from N- to C-termini. The presence of tdTomato enables visualization of transfected cells in the absence of Ca2+ signaling and concentration-independent, ratiometric readout of Ca2+ signals. Salsa6f has been introduced into transgenic mice, where its expression is under Cre recombinase control (see PMID: 29239725).