pcDNA3-blueCKAR(T/A)
(Plasmid #118473)

• Purpose: Negative-control mutant for blueCKAR biosensor.
• Depositing Lab: Jin Zhang
• Publication: Mehta et al Nat Cell Biol. 2018 Sep 24. pii: 10.1038/s41556-018-0200-6. doi: 10.1038/s41556-018-0200-6.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5517
• Total vector size (bp): 6753
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: blueCKAR(T/A)
• Species: Synthetic
• Insert Size (bp): 1236
• Mutation: Contains Thr-to-Ala mutation in substrate sequence.
• Promoter: CMV
• Tags / Fusion Proteins:
  • 6xHIS (N terminal on backbone)
  • T7 tag (gene 10 leader) (N terminal on backbone)
  • Xpress (TM) tag (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • pcDNA3-blueCKAR(TA).gb

Terms and Licenses

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