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pET15b+PCNA3 R112C/T180C
(Plasmid #66182)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 66182 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pET-15b(+)
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5698
  • Total vector size (bp) 6485
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 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
    DNA polymerase sliding clamp 3
  • Alt name
    PCNA3
  • Species
    Synthetic; Sulfolobus solfataricus P2
  • Insert Size (bp)
    798
  • Mutation
    R112C, T180C
  • GenBank ID
    NP_341944
  • Promoter T7 promoter
  • Tag / Fusion Protein
    • His6 tag (N 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 Terminal
    • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

The PCNA3 gene was subcloned from a synthetic gene whose codons are optimized for E. coli expression.

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:

    pET15b+PCNA3 R112C/T180C was a gift from Teruyuki Nagamune (Addgene plasmid # 66182 ; http://n2t.net/addgene:66182 ; RRID:Addgene_66182)

  • For your References section:

    Introduction of selective intersubunit disulfide bonds into self-assembly protein scaffold to enhance an artificial multienzyme complex's activity. Hirakawa H, Kakitani A, Nagamune T. Biotechnol Bioeng. 2013 Jul;110(7):1858-64. doi: 10.1002/bit.24861. Epub 2013 Feb 22. 10.1002/bit.24861 PubMed 23404255
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