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pCMVhTRIM65_3Flag
(Plasmid #53966)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 53966 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pCMV-3Tag-8
  • Backbone manufacturer
    Agilent (formerly Stratagene)
  • Backbone size w/o insert (bp) 5171
  • Total vector size (bp) 6725
  • Modifications to backbone
    Contruct: [XhoI site]CTCGAG-CTG-[HindIII]AAGCTT-(Kozak seq]GCC-insert[hTRIM65 CDS without stop, 1551bp]-[Clai]ATCGAT. The commercial multiple cloning site has been replaced as indicated. Backbone's 3xFLAGs are in frame with the restriction sites.
  • Vector type
    Mammalian Expression
  • Selectable markers
    Hygromycin

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    TRIM65
  • Alt name
    tripartite motif containing 65
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1554
  • GenBank ID
    BC021259.2 has same CDS.
  • Entrez Gene
    TRIM65 (a.k.a. 4732463G12Rik)
  • Promoter CMV
  • Tag / Fusion Protein
    • 3x FLAG (C terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site HindIII (not destroyed)
  • 3′ cloning site Clai (not destroyed)
  • 5′ sequencing primer CMV, T3 (28bp from insert)
  • 3′ sequencing primer T7, BGH reverse
  • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses

Depositor Comments

Cloned from human T98 glioblastoma line. The coding sequence is identical to that in BC021259.2

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:

    pCMVhTRIM65_3Flag was a gift from Martin Dorf (Addgene plasmid # 53966 ; http://n2t.net/addgene:53966 ; RRID:Addgene_53966)
  • For your References section:

    TRIM65 regulates microRNA activity by ubiquitination of TNRC6. Li S, Wang L, Fu B, Berman MA, Diallo A, Dorf ME. Proc Natl Acad Sci U S A. 2014 May 13;111(19):6970-5. doi: 10.1073/pnas.1322545111. Epub 2014 Apr 28. 10.1073/pnas.1322545111 PubMed 24778252