pET28a-CohIII-CBM-HIS-LPETGG
(Plasmid #91697)

• Purpose: CohIII-CBM handle with C-Terminal Sortase Recognition Sequence LPETGG for Posttranslational Coupling to a N-terminal GGG
• Depositing Lab: Hermann Gaub
• Publication: Ott et al ACS Nano. 2017 Jun 12. doi: 10.1021/acsnano.7b02694.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5215
• Total vector size (bp): 6506
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For protein production transformed NiCo21(DE3) strains in auto induction media were grown for 24 hrs at 37°C.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: CohIII-CBM-HIS-LPETGG
• Alt name: Carbohydrate Binding Module
• Alt name: Cohesin Type III
• Species: Synthetic
• Insert Size (bp): 1290
• Mutation: Cysteine deletion in CBM C63S
• Promoter: T7
• Tags / Fusion Proteins:
  • Sortase-Tag (C terminal on insert)
  • HIS-Tag (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CCGATCTTCCCCATCGGT
• 3′ sequencing primer: CTAGTTATTGCTCAGCGGT

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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