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PurposeSalsa6f is a fluorescent ratiometric genetically encoded calcium indicator, (tdTomato linked to GCaMP6f by a V5 epitope tag)
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Depositing Lab
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Depositing OrganizationUniversity of California, Irvine
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 140188 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 140188-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 140188-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFP-N1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 3923
- Total vector size (bp) 6698
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Modifications to backboneThe EGFP gene has been replaced with the ratiometric genetically encoded calcium indicator Salsa6f (tdTomato-V5-GCaMP6f)
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameSalsa6f
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Alt nametdTomato-v5-GCaMP6f
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SpeciesSynthetic
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Insert Size (bp)2775
- Promoter CMV Immediate Early
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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
- (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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).
DNA (Catalog # 140188-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
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Academic/Nonprofit Terms
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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 # 140188-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
-
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
Salsa6f was a gift from Michael Cahalan (Addgene plasmid # 140188 ; http://n2t.net/addgene:140188 ; RRID:Addgene_140188) -
For your References section:
T-cell calcium dynamics visualized in a ratiometric tdTomato-GCaMP6f transgenic reporter mouse. Dong TX, Othy S, Jairaman A, Skupsky J, Zavala A, Parker I, Dynes JL, Cahalan MD. eLife. 2017 Dec 14;6. pii: 32417. doi: 10.7554/eLife.32417. 10.7554/eLife.32417 PubMed 29239725