pET24a-VHH-tev-mCherry
(Plasmid #117752)

• Purpose: Bacterial expression of a functionalized anti-GFP (VHH) nanobody fused to mCherry. VHH-mCherry also contains a T7, HA, BAP, tev cleavage site and His6 epitope
• Depositing Lab: Martin Spiess
• Publication: Buser et al J Vis Exp. 2019 Feb 21;(144). doi: 10.3791/59111.

Backbone

• Vector backbone: pET24a
• Backbone manufacturer: Merck - Novagen
• Total vector size (bp): 6501
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: anti-GFP nanobody fused to a T7, tev site, mCherry, HA, BAP and His6 epitope
• Alt name: VHHGFP4
• Alt name: mCherry
• Species: Synthetic
• Insert Size (bp): 1263
• Promoter: T7
• Tags / Fusion Proteins:
  • T7 (N terminal on insert)
  • HA (C terminal on insert)
  • BAP (C terminal on insert)
  • His6 (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: BamHi (not destroyed)
• 5′ sequencing primer: TAATACGACTCACTATAGGG
• 3′ sequencing primer: TGCTAGTTATTGCTCAGCGG

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry