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pcDNA4/TO-bio-myc-cCE
(Plasmid #112711)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 112711 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pcDNA4/TO
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5078
  • Total vector size (bp) 7203
  • Vector type
    Mammalian Expression
  • Selectable markers
    Zeocin

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
    Rngtt
  • Alt name
    CE, mCE
  • Alt name
    Capping enzyme
  • Alt name
    RNA guanylyltransferase and 5'-phosphatase
  • Species
    M. musculus (mouse)
  • Insert Size (bp)
    2189
  • Mutation
    Silent mutation at Arg 132 (CGT to CGC), deleted amino acids 573-576 (KRKY nuclear localization signal)
  • Entrez Gene
    Rngtt (a.k.a. AU020997, HCE, MCE1)
  • Promoter CMV with Tet repressor binding sites
  • Tags / Fusion Proteins
    • biotinylation signal peptide (Tagwerker et al. 2006 Mol Cell Proteomics, 10.1074/mcp.M500368-MCP200) (N terminal on insert)
    • myc tag (N terminal on insert)
    • HIV Rev nuclear localization signal (NES) (N terminal on insert)
    • Stop codon followed by an out-of-frame FLAG tag (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site KpnI (not destroyed)
  • 3′ cloning site ApaI (not destroyed)
  • 5′ sequencing primer pcDNA4TO-CMV-F (CGCAAATGGGCGGTAGGCGTG)
  • 3′ sequencing primer pcDNA4TO-R (AGCATGCCTGCTATTGTCTTCCCA)
    • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Source of Rngtt sequence: pCR21-mCE, provided by Aaron Shatkin, Rutgers University
  • Article Citing this Plasmid

Terms and Licenses

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

Depositor Comments

For more information about this plasmid, see Mukherjee et al. 2014 PLoS Biol (10.1371/journal.pbio.1001933). The insert sequence of this plasmid was subcloned from pcDNA3-bio-myc-NES-mCEΔNLS described in this publication.

Additional internal sequencing primers:
IntCE1-F: GAATGCCCCACCACTGAGAATACTG
IntCE2-F: GTTAGGAGAGGTGCAGCAGAAATGTC
IntCE3-F: CAAAGAAGTCAGCCATGAAATGGATGGAC

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:

    pcDNA4/TO-bio-myc-cCE was a gift from Daniel Schoenberg (Addgene plasmid # 112711 ; http://n2t.net/addgene:112711 ; RRID:Addgene_112711)

  • For your References section:

    RNA-binding proteins and heat-shock protein 90 are constituents of the cytoplasmic capping enzyme interactome. Trotman JB, Agana BA, Giltmier AJ, Wysocki VH, Schoenberg DR. J Biol Chem. 2018 Oct 26;293(43):16596-16607. doi: 10.1074/jbc.RA118.004973. Epub 2018 Aug 30. 10.1074/jbc.RA118.004973 PubMed 30166341
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