pET28a- mCherry-FIVAR-HIS-ybbR
(Plasmid #169138)

• Purpose: E. coli expression of mCherry domain containing ybbR tag and FIVAR 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): 6272
• 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: mCherry
• Alt name: monomeric Cherry
• Species: Discosoma sp.
• Insert Size (bp): 1050
• Tags / Fusion Proteins:
  • FIVAR tag (C terminal on insert)
  • 6x Histag (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

• Supplemental Documents:
  • pET28a_mCherry-FIVAR-HIS-ybbR.gb
• A portion of this plasmid was derived from a plasmid made by: This plasmid was derived from a pBAD vector encoding mCherry made by Michael Davidson & Nathan Shaner & Roger Tsien (Addgene plasmid # 54630 ; http://n2t.net/addgene:54630 ; RRID:Addgene_54630).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• 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.