pMBP-SpyCatcher
(Plasmid #79053)

• Purpose: Expresses maltose binding protein (MBP) SpyCatcher fusion
• Depositing Lab: James Bowie
• Publication: Min et al Protein Sci. 2016 Aug;25(8):1535-44. doi: 10.1002/pro.2952. Epub 2016 Jun 6.

Backbone

• Vector backbone: pDEST14
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 3644
• Total vector size (bp): 5168
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: MBP-SpyCatcher
• Species: Streptococcus pyogenes
• Insert Size (bp): 1524
• Mutation: Derived from pDEST14-SpyCatcher (Plasmid #35044); deletion of C-terminal till I113, and point mutations I34E and M69Y
• Promoter: T7
• Tags / Fusion Proteins:
  • Maltose binding protein (N terminal on insert)
  • 6xHis (N terminal on insert)

Cloning Information

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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Howarth lab - plasmid is a derivative of pDEST14-SpyCatcher (Plasmid #35044)

Terms and Licenses

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

Depositor Comments

Transform plasmid into E. coli BL21(DE3) for expression. Use amylose resin for purification; His6 tag is partially inaccessible and not suitable for purification.