Orai1-jGCaMP8f V203Y
(Plasmid
#231631)
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PurposeGreen fluorescent reporter of Orai1-associated calcium influx
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 231631 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepEGFP-N1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 3958
- Total vector size (bp) 6169
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Modifications to backboneThe EGFP gene has been replaced with the genetically encoded calcium indicator jGCaMP8f V203Y.
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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 nameOrai1-jGCaMP8f V203Y
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SpeciesH. sapiens (human), Synthetic
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Insert Size (bp)2211
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Entrez GeneORAI1 (a.k.a. CRACM1, IMD9, ORAT1, TAM2, TMEM142A)
- Promoter CMV Immediate Early
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Tag
/ Fusion Protein
- jGCaMP8f V203Y (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site XhoI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer TGACGTCAATGGGAGTTTG
- 3′ sequencing primer ACATTGATGAGTTTGGAC (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made bythe GENIE Project, Janelia Research Campus, plasmid pGP-CMV-jGCaMP8f (Addgene #162373) containing jGCaMP8f.
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 contains the genetically encoded calcium indicator jGCaMP8f from the plasmid pGP-CMV-jGCaMP8f (Addgene plasmid # 162373). jGCaMP8f has been mutated (V203Y, EGFP numbering) to minimize fluorophore photoswitching. Note that the hOrai1 gene isolated in the Cahalan lab contains a silent mutation/polymorphism (T798C; aa T266).
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:
Orai1-jGCaMP8f V203Y was a gift from Michael Cahalan (Addgene plasmid # 231631 ; http://n2t.net/addgene:231631 ; RRID:Addgene_231631) -
For your References section:
Photoswitching alters fluorescence readout of jGCaMP8 Ca(2+) indicators tethered to Orai1 channels. Dynes JL, Yeromin AV, Cahalan MD. Proc Natl Acad Sci U S A. 2023 Sep 26;120(39):e2309328120. doi: 10.1073/pnas.2309328120. Epub 2023 Sep 20. 10.1073/pnas.2309328120 PubMed 37729200