pet30b-5M SrtA
(Plasmid #51140)

• Purpose: expression plasmid for C-terminal his tagged S. aureus SrtA with enhanced catalytic activity
• Depositing Lab: Hidde Ploegh
• Publication: Shi et al Proc Natl Acad Sci U S A. 2014 Jul 15;111(28):10131-6. doi: 10.1073/pnas.1409861111. Epub 2014 Jun 30.

Backbone

• Vector backbone: pET30b
• Backbone manufacturer: Novagen
• 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: S.aureus SrtA 5M
• Alt name: Sortase A pentamutant
• Species: S.aureus
• Insert Size (bp): 471
• Mutation: P94R, D160N, D165A, K190E, K196T
• Promoter: T7
• Tag / Fusion Protein:
  • 6x His (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7-term

Resource Information

• Articles Citing this Plasmid:
  • 7 References

Terms and Licenses

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

Depositor Comments

Pentamutant sortase A was originally described by David R. Liu's laboratory (Harvard University) in:
A General Strategy for the Evolution of Bond-Forming Enzymes Using Yeast Display. Chen I, Dorr BM, Liu DR. Proc Natl Acad Sci U S A. 2011 Jul 12;108(28):11399-404 (PubMed ID: 21697512)