pD-MSP1D1
(Plasmid #161607)

• Purpose: Bacterial expression of mutated membrane scaffold protein ("dark" MSP1D1) for Nanodisc formation
• Depositing Lab: Stephen Sligar
• Publication: McLean et al Anal Biochem. 2020 Oct 15;607:113860. doi: 10.1016/j.ab.2020.113860. Epub 2020 Aug 1.

Backbone

• Vector backbone: pET 28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5300
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: for protein expression use strain with internal source of T7 polymerase e.g. BL21(DE3)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: D-MSP1D1
• Alt name: membrane scaffold protein
• Alt name: APOA1
• Alt name: PDB: 1AV1
• Species: synthetic gene
• Insert Size (bp): 640
• Mutation: mutant of MSP1D1; all tryptophans were replaced by phenylalanines
• Promoter: T7
• Tag / Fusion Protein:
  • 7-His tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Nco 1 (not destroyed)
• 3′ cloning site: Hind III (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7 term

Terms and Licenses

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