Orai1-GCaMP6f
(Plasmid #73564)

• Purpose: Green fluorescent reporter of Orai1-associated calcium influx
• Depositing Lab: Michael Cahalan
• Publication: Dynes et al Proc Natl Acad Sci U S A. 2016 Jan 12;113(2):440-5. doi: 10.1073/pnas.1523410113. Epub 2015 Dec 28.

Backbone

• Vector backbone: pEGFP-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3958
• Total vector size (bp): 6166
• Modifications to backbone: The EGFP gene has been replaced with the genetically encoded calcium indicator 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: Orai1-GCaMP6f
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 2208
• GenBank ID: NM_032790.3
• Entrez Gene: ORAI1 (a.k.a. CRACM1, IMD9, ORAT1, TAM2, TMEM142A)
• Promoter: CMV Immediate Early
• Tag / Fusion Protein:
  • GCaMP6f (C terminal on insert)

Cloning Information

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

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The fusion protein in this plasmid contains the genetically encoded calcium indicator GCaMP6f from the plasmid pGP-CMV-GCaMP6f (Addgene plasmid # 40755) provided by Douglas Kim. Note that the hOrai1 gene isolated in the Cahalan lab contains a silent mutation/polymorphism (T798C; aa T266).