pET28a-ybbR-HIS-Xyl-DocI
(Plasmid #58711)

• Purpose: Characteristic fingerprint molecule to perform single molecule force spectroscopy experiments; covalently immobilized via ybbr tag
• Depositing Lab: Hermann Gaub
• Publication: Otten et al Nat Methods. 2014 Sep 7. doi: 10.1038/nmeth.3099.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5216
• Total vector size (bp): 6689
• 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: Xyl-DocI
• Alt name: Xylanase T6
• Alt name: dockerin type I
• Species: Synthetic; Geobacillus stearothermophilus, Clostridium thermocellum
• Insert Size (bp): 1473
• Mutation: changed Threonin 120 to Cysteine in Xylanase Domain
• Promoter: T7
• Tags / Fusion Proteins:
  • ybbR (N terminal on insert)
  • HIS (N terminal on insert)
  • HRV 3C (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: We thankfully received the xylanase and dockerin domain from Professor Edward Bayer (Department of Biological Chemistry, Weizmann Institute of Science, Israel) as recently described in this paper: Stahl, S. W., Nash, M. A., Fried, D. B., Slutzki, M., Barak, Y., Bayer, E. A., & Gaub, H. E. (2012). Single-molecule dissection of the high-affinity cohesin-dockerin complex. Proceedings of the National Academy of Sciences of the United States of America, 109(50), 20431–6. doi:10.1073/pnas.1211929109

Terms and Licenses

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