SpyTag-β-Lactamase-SpyCatcher
(Plasmid #52656)

• Purpose: Expresses SpyTag-SpyCatcher mediated cyclized version of TEM-1 β-lactamase (BLA). Cyclization of BLA led to an increase in thermal resilience of >60 degrees celsius.
• Depositing Lab: Mark Howarth
• Publication: Schoene et al Angew Chem Int Ed Engl. 2014 Jun 10;53(24):6101-4. doi: 10.1002/anie.201402519. Epub 2014 May 9.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5369
• Total vector size (bp): 6532
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: TEM-1 β-Lactamase
• Alt name: BLA
• Species: Synthetic; E. coli
• Insert Size (bp): 1284
• GenBank ID: YP_006952162.1 KJ645919
• Promoter: T7 promoter
• Tags / Fusion Proteins:
  • SpyTag (N terminal on insert)
  • SpyCatcher (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: T7F
• 3′ sequencing primer: T7R

Resource Information

• Supplemental Documents:
  • Instructions on how to easily insert your favorite protein into the SpyTag-SpyCatcher scaffold
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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