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3XFlagNICD2
(Plasmid #20184)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 20184 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    p3XFLAG-CMV-7
  • Backbone manufacturer
    Sigma
  • Backbone size w/o insert (bp) 4700
  • Vector type
    Mammalian Expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Notch 2 intracellular domain
  • Alt name
    NICD2
  • Alt name
    Notch 2
  • Species
    M. musculus (mouse)
  • Insert Size (bp)
    2403
  • Mutation
    Contains murine Notch 2 Intracellular Domain (Met-1699 to Ala-2470)
  • GenBank ID
    NM_010928.2 NP_035058.2
  • Entrez Gene
    Notch2 (a.k.a. AI853703, N2)
  • Promoter CMV
  • Tag / Fusion Protein
    • 3X FLAG (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Xba1 (not destroyed)
  • 3′ cloning site BamH1 (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer hGH-pA-R
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    Notch 2 plasmid from Dr. Yoshio Hamada (NIBB, Japan)
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
  • Articles Citing this Plasmid

Depositor Comments

All NICD plasmids were cloned into the p3XFLAG-CMV-7(TM) expression vector (Sigma). NICD2 was cloned into XbaI/BamHI sites. NICD2 (Met-1699 to Ala-2470 of full-length mouse N2) was amplified from the N2 plasmid (gift from Dr. Y. Hamada) using primers 5XbaN2 (5'-GCTCTAGAATGGCCAAGCGGAAGCGCAAG-3') and 3BamN2
(5'-GCGGATCCCTATGCATACACCTGCATGTTGCTG-3').

Please note that Addgene sequencing results confirm the updated domain numbering to be Met 1701-Ala 2473.

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:

    3XFlagNICD2 was a gift from Raphael Kopan (Addgene plasmid # 20184 ; http://n2t.net/addgene:20184 ; RRID:Addgene_20184)
  • For your References section:

    Target selectivity of vertebrate notch proteins. Collaboration between discrete domains and CSL-binding site architecture determines activation probability. Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan R. J Biol Chem. 2006 Feb 24. 281(8):5106-19. 10.1074/jbc.M506108200 PubMed 16365048