GLuc-FLAG-CB1
(Plasmid #158066)

• Purpose: Membrane expression detection of Cannabinoid receptor 1
• Depositing Lab: Adolfo Rivero-Muller
• Publication: Rodriguez-Rodriguez et al Sci Rep. 2020 Oct 23;10(1):18191. doi: 10.1038/s41598-020-75331-y.

Backbone

• Vector backbone: PX459v2
• Backbone manufacturer: Feng Zhang's Lab
• Backbone size w/o insert (bp): 9175
• Total vector size (bp): 10146
• Modifications to backbone: Promoter: chicken β-actin promoter with CMV enhancer
• Vector type: Mammalian Expression, AAV

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: CB1
• Alt name: CNR1 cannabinoid receptor 1
• Species: H. sapiens (human)
• Insert Size (bp): 5996
• Mutation: contains 4Kb of 3' UTR
• GenBank ID: 1268
• Entrez Gene: CNR1 (a.k.a. CANN6, CB-R, CB1, CB1A, CB1K5, CB1R, CNR)
• Promoter: pCAG
• Tag / Fusion Protein:
  • Gaussia Luciferase (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ggctgtaattagctgagcaagagg
• 3′ sequencing primer: GCTGTCCTGCCCCACCCCAC

Terms and Licenses

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

Depositor Comments

Note that the Addgene verified sequence contains a few ambiguous nucleotides in the CAG promoter region. These discrepancies do not impact plasmid function and are likely due to difficulties sequencing GC-rich regions. The N-terminus of GLuc-Flag-CB1 contains additional residues (MIKIATRKYLGKQNVYDIGVERDHNFALKNGFIASNCFN) which match DnaE-C (NCBI reference WP_114085349.1).