pNCS mCherry
(Plasmid #91769)

• Purpose: Bacterial expression of a fluorescent protein, monomeric Cherry (mCherry)
• Depositing Labs: Erik Rodriguez, Roger Tsien
• Publication: Shaner et al Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21.

Backbone

• Vector backbone: modified pRSETB
• Backbone manufacturer: Life Technologies
• 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: monomeric Cherry
• Alt name: mCherry
• Species: Discosoma sp. (mushroom coral)
• Insert Size (bp): 735
• Mutation: See Depositor Comments
• Promoter: T7
• Tag / Fusion Protein:
  • Hexa-Histidine tag, 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 (not destroyed)
• 3′ cloning site: EcoRI (not 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
  • Takara Bio Limited Use Label License (formerly Clontech)
• 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.

Addgene next-generation sequencing identified a deletion of Glutimate and Glycine residues (amino acids 73 and 74 relative to Genbank ID: 6MZ3_A). The depositing laboratory has observed expression of the chromophore in E. coli.