MO70 - Orai1 EC-Flag
(Plasmid #79590)

• Purpose: Transient expression and retroviral Expression
• Depositing Lab: Yousang Gwack
• Publication: Srikanth et al J Biol Chem. 2010 Feb 12;285(7):5066-75. doi: 10.1074/jbc.M109.072736. Epub 2009 Dec 10.

Backbone

• Vector backbone: MO70
• Backbone size w/o insert (bp): 7480
• Vector type: Mammalian Expression, Retroviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Orai1
• Alt name: TMEM142A
• Alt name: CRACM1
• Species: H. sapiens (human)
• Insert Size (bp): 901
• Entrez Gene: ORAI1 (a.k.a. CRACM1, IMD9, ORAT1, TAM2, TMEM142A)
• Promoter: CMV
• Tag / Fusion Protein:
  • FLAG (inserted between TM3 and TM4 of the protein)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: pCDH-Rev

Terms and Licenses

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

Depositor Comments

This plasmid contains a FLAG tag in its extracellular loop 2 between transmembranes 3 and 4. This plasmid can be used for surface staining to detect Orai1 expression on the plasma membrane.

Please note that compared to GenBank reference sequence NP_116179.2, the Orai1 insert in this plasmid is missing two amino acids (P8 and P9). This difference does not affect the function, as the clone has been tested for functionality in Orai1 KO MEFs and can reconstitute Orai1 function in Orai1 KO cells identical to WT (untagged) Orai1. The functional reconstitution data are presented in the original JBC paper (PMID 20007711) describing this construct.