Skip to main content
Addgene

pKM586
(Plasmid #19978)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 19978 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pZS*24-MCS1
  • Backbone manufacturer
    Herman Bujard
  • Backbone size w/o insert (bp) 4300
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    BL21-Pro
  • Growth instructions
    30 degrees
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    TEV protease
  • Species
    tobacco etch virus
  • Insert Size (bp)
    900
  • Mutation
    S219D mutant
  • Entrez Gene
    TEVgp1 (a.k.a. TEVgp1)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer n/a
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    A pZ derivative from - Lutz, R. and Bujard, H. (1997). Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucl. Acids Res. 25: 1203-10.

Terms and Licenses

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

Depositor Comments

This TEV protease expression vector is intended to be used for controlled intracellular processing of fusion proteins (TEV protease substrates) in E. coli. It is supplied in BL21-Pro cells so that transcription of TEV protease can be regulated by anhydrotetracycline.

Replicon: pSC101
Promoter: Synthetic ªPL/tetO

Inducer: anhydrotetracycline (100 ºg/liter)
Notes: Expression of the TEV protease gene on pKM586 will be constitutive unless the host strain produces Tet repressor. Regulated expression can be achieved in DH5alphaPro or BL21Pro cells (Clontech). For best results, perform intracellular processing experiments at 30 °C.

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:

    pKM586 was a gift from David Waugh (Addgene plasmid # 19978 ; http://n2t.net/addgene:19978 ; RRID:Addgene_19978)
  • For your References section:

    Controlled intracellular processing of fusion proteins by TEV protease. Kapust RB, Waugh DS. Protein Expr Purif. 2000 Jul . 19(2):312-8. 10.1006/prep.2000.1251 PubMed 10873547