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Addgene

pMT643
(Plasmid #8610)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 8610 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pUC19
  • Backbone size w/o insert (bp) 2641
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    barstar A
  • Species
    Bacillus amyloliquefaciens
  • Insert Size (bp)
    439
  • Mutation
    Two cysteines of barstar changed to alanine, C40A and C82A
  • GenBank ID
    X15545

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site EcoRI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer M13F
  • 3′ sequencing primer M13R
  • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

As pMT316 but with the TGT codons for the two cysteines of barstar, C40 and C82, changed to GCG for alanine. The protein product, barstar A, has no sulfhydryl groups but is functional in vivo and in vitro, with only slight reductions in barnase binding and stability.

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:

    pMT643 was a gift from Robert Hartley (Addgene plasmid # 8610 ; http://n2t.net/addgene:8610 ; RRID:Addgene_8610)
  • For your References section:

    Directed mutagenesis and barnase-barstar recognition. Hartley RW. Biochemistry 1993 Jun 15;32(23):5978-84. 10.1021/bi00074a008 PubMed 8507637