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Addgene

His6-MBP-uTEV3
(Plasmid #135464)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 135464 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pYFJ16
  • Backbone manufacturer
    John Cronan
  • Backbone size w/o insert (bp) 4857
  • Total vector size (bp) 6785
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Use BL21(DE3)-RIL for protein expression. Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble TEV protease. Ampicillin for His6-MBP-uTEV3; Chloramphenicol for pRIL.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    His6-MBP-uTEV3
  • Species
    Synthetic
  • Mutation
    S219V mutation (improves stability) , I138T/S153N/T180A mutations improve activity. Last 6 amino acids from TEV were deleted (237-242)
  • Promoter T5
  • Tag / Fusion Protein
    • MBP, His6

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer GCTTTGTGAGCGGATAACAATTATAATAG
  • 3′ sequencing primer CTGGCAAACGGTCGCCAGAGATCCGCG
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.
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:

    His6-MBP-uTEV3 was a gift from Alice Ting (Addgene plasmid # 135464 ; http://n2t.net/addgene:135464 ; RRID:Addgene_135464)
  • For your References section:

    Directed evolution improves the catalytic efficiency of TEV protease. Sanchez MI, Ting AY. Nat Methods. 2019 Dec 9. pii: 10.1038/s41592-019-0665-7. doi: 10.1038/s41592-019-0665-7. 10.1038/s41592-019-0665-7 PubMed 31819267