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Addgene

pET28b+R3(R112C/T180C)C
(Plasmid #108194)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 108194 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET-28b(+)
  • Backbone manufacturer
    Merck Millipore
  • Backbone size w/o insert (bp) 5271
  • Total vector size (bp) 8666
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    Supplement with 1% glucose is recommended to suppress leaky expression.
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    PdR-PCNA3(R112C/T180C)-P450cam fusion protein
  • Species
    Synthetic; Pseudomonas putida, Sulfolobus solfataricus P2
  • Insert Size (bp)
    3395
  • Mutation
    R112C/T180C(PCNA3)
  • Promoter T7 promoter
  • Tag / Fusion Protein
    • His6 tag (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NcoI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer T7
  • 3′ sequencing primer T7 Terminal
    • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Masahiro Goto, Kyushu University; Stephen Sligar, University of Illinois at Urbana-Champaign (pUS200, Addgene plasmid # 17788)

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:

    pET28b+R3(R112C/T180C)C was a gift from Teruyuki Nagamune (Addgene plasmid # 108194 ; http://n2t.net/addgene:108194 ; RRID:Addgene_108194)

  • For your References section:

    Artificial Self-Sufficient Cytochrome P450 Containing Multiple Auxiliary Proteins Demonstrates Improved Monooxygenase Activity. Haga T, Hirakawa H, Nagamune T. Biotechnol J. 2018 Dec;13(12):e1800088. doi: 10.1002/biot.201800088. Epub 2018 Aug 14. 10.1002/biot.201800088 PubMed 30039932
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