pET28a-VHH-ddFLN4-HIS-ybbR
(Plasmid #169137)

• Purpose: E. coli expression of VHH domain containing ybbR tag and ddFLN4 fingerprint domain. This construct was designed for AFM-SMFS measurements.
• Depositing Lab: Michael Nash
• Publication: Liu et al JACS Au. 2022 May 18;2(6):1417-1427. doi: 10.1021/jacsau.2c00121. eCollection 2022 Jun 27.

Backbone

• Vector backbone: pET28a
• Backbone size w/o insert (bp): 5222
• Total vector size (bp): 6014
• 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: VHH
• Alt name: mCherry-Nanobody
• Alt name: LaM-4
• Species: Synthetic
• Insert Size (bp): 792
• Tags / Fusion Proteins:
  • ddFLN4 tag (C terminal on insert)
  • 6x His tag (C terminal on insert)
  • ybbr tag (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Resource Information

• A portion of this plasmid was derived from a plasmid made by: mCherry-nanobody (LaM-4) was originally developed by Dr. Rout et al. (Fridy PC, Li Y, Keegan S, Thompson MK, Nudelman I, Scheid JF, Oeffinger M, Nussenzweig MC, Fenyö D, Chait BT & Rout MP (2014) A robust pipeline for rapid production of versatile nanobody repertoires. Nat Meth 11: 1253–1260)

Terms and Licenses

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

Depositor Comments

Additional citation: Liu, H., Schittny, V. and Nash, M.A., 2019. Removal of a conserved disulfide bond does not compromise mechanical stability of a VHH antibody complex. Nano letters, 19(8), pp.5524-5529.