Skip to main content
Addgene

Orai1-jGCaMP8f V203Y
(Plasmid #231631)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 231631 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pEGFP-N1
  • Backbone manufacturer
    Clontech
  • Backbone size w/o insert (bp) 3958
  • Total vector size (bp) 6169
  • Modifications to backbone
    The EGFP gene has been replaced with the genetically encoded calcium indicator jGCaMP8f V203Y.
  • 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
    Orai1-jGCaMP8f V203Y
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    2211
  • Entrez Gene
    ORAI1 (a.k.a. CRACM1, IMD9, ORAT1, TAM2, TMEM142A)
  • Promoter CMV Immediate Early
  • 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

  • A portion of this plasmid was derived from a plasmid made by
    the GENIE Project, Janelia Research Campus, plasmid pGP-CMV-jGCaMP8f (Addgene #162373) containing jGCaMP8f.

Terms and Licenses

  • Academic/Nonprofit Terms
  • 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).

How to cite this plasmid ( Back to top)

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.

  • 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