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eSrtA(4S-9) pET29b
(Plasmid #75146)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 75146 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET29b
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5370
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    reprogrammed sortase A variant
  • Alt name
    eSrtA 4S-9
  • Species
    Staphylococcus aureus
  • Insert Size (bp)
    441
  • Mutation
    N98D S102C A104V A118T F122A K134R F144L I182V E189F compared to eSrtA. Deleted amino acids 1-59 (which do not affect catalytic activity).
  • 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 BamHI (not destroyed)
  • 5′ sequencing primer T7
  • 3′ sequencing primer T7-term
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

eSrtA(4S-9) has 120 fold change in substrate specificity with activity comparable to eSrtA and it prefers LPXSG substrate over wild-type LPXTG. See sortase A pentamutant (eSrtA) in pET29 for original eSrtA (Addgene plasmid# 75144).

How to cite this plasmid ( Back to top)

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:

    eSrtA(4S-9) pET29b was a gift from David Liu (Addgene plasmid # 75146 ; http://n2t.net/addgene:75146 ; RRID:Addgene_75146)
  • For your References section:

    Reprogramming the specificity of sortase enzymes. Dorr BM, Ham HO, An C, Chaikof EL, Liu DR. Proc Natl Acad Sci U S A. 2014 Sep 16;111(37):13343-8. doi: 10.1073/pnas.1411179111. Epub 2014 Sep 3. 10.1073/pnas.1411179111 PubMed 25187567