pET28a-HIS-Xyl-Xmod-DocIII
(Plasmid #60865)

• Purpose: Characteristic fingerprint molecule fused to dockerin type III domain to perform single molecule force spectroscopy experiments; covalently immobilized via introduced Cystein in Xylanase T120C
• Depositing Lab: Hermann Gaub
• Publication: Schoeler et al Nat Commun. 2014 Dec 8;5:5635. doi: 10.1038/ncomms6635.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 4304
• Total vector size (bp): 6194
• 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 1

• Gene/Insert name: Xyl
• Alt name: Xylanase
• Species: Synthetic; Bacillus stearothermophilus
• Insert Size (bp): 1161
• Mutation: Threonine 120 to Cysteine in Xylanase domain
• Promoter: T7
• Tag / Fusion Protein:
  • 6xHIS (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BspHI (destroyed during cloning)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Reverse

Gene/Insert 2

• Gene/Insert name: Xmod-DocIII
• Alt name: dockerin type III
• Alt name: x-module
• Species: Ruminococcus flavefaciens
• Insert Size (bp): 729
• Promoter: T7

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 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 plasmid from Professor Edward Bayer (Department of Biological Chemistry, Weizmann Institute of Science, Israel) as recently described in this paper: "Atypical cohesin-dockerin complex responsible for cell surface attachment of cellulosomal components: binding fidelity, promiscuity, and structural buttresses."Salama-Alber Orly, Jobby Maroor K., Chitayat Seth, Smith Steven P., White Bryan A., Shimon Linda J. W., Lamed Raphael, Frolow Felix, Bayer Edward A. (The Journal of biological chemistry, 2013, 288 (23).

Terms and Licenses

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