pNCS BFP
(Plasmid #91757)

• Purpose: Bacterial expression of a fluorescent protein, Blue Fluorescent Protein (BFP)
• Depositing Labs: Erik Rodriguez, Roger Tsien
• Publication: Cubitt et al Trends Biochem Sci. 1995 Nov;20(11):448-55.

Backbone

• Vector backbone: modified pRSETB
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2900
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: E. coli expressing T7 polymerase necessary for fluorescent protein production. Expresses fluorescent protein constitutively. Do not add IPTG.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Blue Fluorescent Protein
• Alt name: BFP
• Species: Aequorea victoria (jellyfish)
• Insert Size (bp): 735
• Mutation: GFP (Y66H)
• Promoter: T7
• Tag / Fusion Protein:
  • Hexa-Histidine, Xpress epitope for detection with the anti-Xpress antibody, and enterokinase cleavage site. (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: EcoRI (unknown if destroyed)
• 5′ sequencing primer: T7 Promoter Forward
• 3′ sequencing primer: T7 Terminator

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

These plasmids are used in University Molecular Biology Laboratories and by High School students to purify fluorescent proteins and create fluorescent protein plate artwork.