sortase A pentamutant (eSrtA) in pET29
Purposeevolved sortase A (eSrtA) pentamutant with improved kinetics and activity
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||75144||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5370
Vector typeBacterial Expression
Growth in Bacteria
Bacterial Resistance(s)Kanamycin, 50 μg/mL
Copy numberHigh Copy
Gene/Insert namesortase A pentamutant
Alt name5M SrtA
Alt nameevolved sortase A (eSrtA)
Insert Size (bp)441
MutationP94R, D160N, D165A, K190E, K196T. Deleted amino acids 1-59 (which do not affect catalytic activity).
- Promoter T7
/ Fusion Protein
- 6x His (C terminal on backbone)
- 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 (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
This plasmid has been found to be somewhat unstable and prone to concatenation. Concatenation often does not impact plasmid function, but may reduce transformation efficiencies. You may need to screen multiple colonies to isolate the monomeric version of this plasmid. If you still have trouble isolating the monomeric version, you might consider linearizing, gel extracting, re-ligating, and transforming the plasmid.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:sortase A pentamutant (eSrtA) in pET29 was a gift from David Liu (Addgene plasmid # 75144 ; http://n2t.net/addgene:75144 ; RRID:Addgene_75144)
For your References section: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. doi: 10.1073/pnas.1101046108. Epub 2011 Jun 22. 10.1073/pnas.1101046108 PubMed 21697512