pNCS mOrange
(Plasmid #91764)

• Purpose: Bacterial expression of a fluorescent protein, monomeric Orange (mOrange)
• 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 Orange
• Alt name: mOrange
• Species: Discosoma sp. (mushroom coral)
• Insert Size (bp): 735
• 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

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.