3XFlagNICD2
(Plasmid #20184)

• Depositing Lab: Raphael Kopan
• Publication: Ong et al J Biol Chem. 2006 Feb 24. 281(8):5106-19.

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, 100 μg/mL
• 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. 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

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Notch 2 plasmid from Dr. Yoshio Hamada (NIBB, Japan)
• Articles Citing this Plasmid:
  • 11 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.