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pPD3cC
(Plasmid #86972)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 86972 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) 5700
  • Total vector size (bp) 8775
  • Modifications to backbone
    A SapI recognition site (GAAGAGC) was modified to GTTGAGC
  • 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
    PCNA3 variant (R112C/T180C) fused to phosphite dehydrogenase and P450cam
  • Alt name
    PTDH-PCNA3R112C/T180C-P450cam
  • Species
    Synthetic; Pseudomonas, Sulfolobus
  • Insert Size (bp)
    3075
  • Mutation
    E175A/A176R in PTDH, N106R/R112C/T180C in PCNA3, silent mutations in P450cam
  • GenBank ID
    AAA25760.1 WP_009988781.1
  • 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)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    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.

Depositor Comments

According to the numbering of NCBI ref WP_009988781.1, the mutations in PCNA3 are N106R, R127C, and T195C.

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:

    pPD3cC was a gift from Teruyuki Nagamune (Addgene plasmid # 86972 ; http://n2t.net/addgene:86972 ; RRID:Addgene_86972)

  • For your References section:

    Supramolecular protein assembly supports immobilization of a cytochrome P450 monooxygenase system as water-insoluble gel. Tan CY, Hirakawa H, Nagamune T. Sci Rep. 2015 Mar 3;5:8648. doi: 10.1038/srep08648. 10.1038/srep08648 PubMed 25733255
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