3XFlagNICD3
(Plasmid #20185)

• 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 3 Intracellular Domain
• Alt name: NICD3
• Alt name: Notch 3
• Species: M. musculus (mouse)
• Insert Size (bp): 1962
• Mutation: Contains murine Notch 3 Intracellular Domain (Met-1664 to Ala-2318)
• GenBank ID: NM_008716.2 NP_032742.1
• Entrez Gene: Notch3 (a.k.a. N3, hpbk)
• 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 3 plasmid from Dr. Urban Lendahl (Karolinska Institutet, Stockholm, Sweden).
• Articles Citing this Plasmid:
  • 5 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). NICD3 was cloned into XbaI/BamHI sites. NICD3 (Met-1664 to Ala-2318 of full-length mouse N3) was amplified from the N3 plasmid (gift from Dr. U. Lendahl) with 5XbaN3 (5'-GCTCTAGAATGGTTGCCAGGCGAAAGCGAG-3'
) and 3BamN3
(5
'-GCGGATCCCTAGGCCATCACCTGCCTCTTGGG-3
') primers.