3XFlagNICD4
(Plasmid #20186)

• 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 4 Intracellular Domain
• Alt name: NICD4
• Alt name: Notch 4
• Species: M. musculus (mouse)
• Insert Size (bp): 1503
• Mutation: Contains murine Notch 4 Intracellular Domain (Val-1463 to Asn-1964)
• GenBank ID: NM_010929.2 NP_035059.2
• Entrez Gene: Notch4 (a.k.a. Int-3, Int3, N4)
• Promoter: CMV
• Tag / Fusion Protein:
  • 3X Flag (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: Xba1 (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-4-containing Int-3 plasmid from Dr. Jan Kitajewski (Columbia University, New York City, New York)
• Articles Citing this Plasmid:
  • 4 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). NICD4 utilized HindIII/XbaI sites. NICD4 (Val-1463 to Asn-1964 of full length mouse N4) was amplified from the Int-3 plasmid (gift from Dr. J. Kitajewski) using primers 5HinN4 (5'-GGCCAAGCTTGTCCTCCAGCTCATTCGGCGACGG-3
') and 3XbaN4
(5
'-GCTCTAGACGGCTTCCGGTGCCTAGTTCAG-3
').