pBAD mApple
(Plasmid #91772)

• Purpose: Bacterial expression of a fluorescent protein, monomeric Apple (mApple). Note: Addgene identified several mutations compared to the final published sequence.
• Depositing Labs: Erik Rodriguez, Roger Tsien
• Publication: Shaner et al Nat Methods. 2008 Jun;5(6):545-51. doi: 10.1038/nmeth.1209. Epub 2008 May 4.

Backbone

• Vector backbone: pBAD
• Backbone manufacturer: Life Technologies
• Backbone size w/o insert (bp): 4000
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: 0.02% (weight/volume) of Arabinose is necessary for fluorescent protein production.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: monomeric Apple
• Alt name: mApple
• Species: Discosoma sp. (mushroom coral)
• Insert Size (bp): 735
• Mutation: R97K, E152S, A180S, V207T compared to mApple (FP Base ID #T5UGO)
• Promoter: araBAD
• 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: pBAD Forward
• 3′ sequencing primer: unecessary

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. Please note that several mutations were identified compared to the final published sequence. It is unknown how these mutations affect the fluorophore's characteristics. Please see plasmids #54536 and 54537 for the published mApple sequence.