pCDNA3.1-nAChr-beta3-CFP
(Plasmid #50485)

• Purpose: this report is the first to directly measure nAChr subunit stoichiometry using FRET and plasma membrane localization of Alpha6 and Beta3 containing receptors using TIRF
• Depositing Lab: Henry Lester
• Publication: Drenan et al Mol Pharmacol. 2008 Jan;73(1):27-41. Epub 2007 Oct 11.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Life Technologies
• Backbone size w/o insert (bp): 5597
• Total vector size (bp): 7827
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: nAChRBeta3 subunit, isoform 1
• Alt name: nAchR beta3-CFP
• Species: M. musculus (mouse)
• Insert Size (bp): 2230
• Mutation: CFP fusion in M3-M4 loop (after residue P379). Our CFP fusion cassette also consisted of a Gly-Ala-Gly flexible linker flanking the CFP open reading frame on both N and C terminal sides
• GenBank ID: AAL75573.1 AF467896_1
• Entrez Gene: Chrnb3 (a.k.a. 5730417K16Rik, Acrb3)
• Promoter: CMV
• Tag / Fusion Protein:
  • CFP

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRV (destroyed during cloning)
• 3′ cloning site: XbaI (unknown if destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Terms and Licenses

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