pET28a_6HT-TBP6.7(R49A)
(Plasmid #112726)

• Purpose: Arg49 of TBP6.7 and other TBPs is the only arginine that was lab-evolved from wild-type U1A to play a significant role in TAR RNA recognition. Mutation results in ~230-fold reduction in Kd.
• Depositing Lab: Joseph Wedekind
• Publication: Belashov et al Nucleic Acids Res. 2018 Jul 27;46(13):6401-6415. doi: 10.1093/nar/gky529.

Backbone

• Vector backbone: pET28a(+)
• Backbone size w/o insert (bp): 5277
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 6His-TEV-TBP6.7(R49A)
• Insert Size (bp): 359
• Mutation: R49A
• Promoter: T7
• Tag / Fusion Protein:
  • 6His-TEV (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Terms and Licenses

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