pmCherry-imp α
(Plasmid #119719)

• Purpose: Expresses mCherry-tagged human importinα in mammalian cells
• Depositing Lab: Kenji Sugimoto
• Publication: Nakagawa et al Biosci Biotechnol Biochem. 2012;76(2):388-90. doi: 10.1271/bbb.110677. Epub 2012 Feb 7.

Backbone

• Vector backbone: pmCherry-C1
• Backbone manufacturer: not commercially available one. Note that Eco47III, AgeI, BspE1 are missing.
• Modifications to backbone: NheI-BglII DNA fragment encoding EGFP of pEGFP is replaced by the DNA fragment encoding mCherry, which is amplified by PCR using pRSET B mCherry (provided by Dr. Roger Y. Tsien).
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: importinα
• Species: H. sapiens (human)
• Mutation: contains amino acids 251-529
• Entrez Gene: KPNA2 (a.k.a. IPOA1, PTAC58, QIP2, RCH1, SRP1-alpha, SRP1alpha)
• Promoter: CMV
• Tag / Fusion Protein:
  • mCherry (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (unknown if destroyed)
• 3′ cloning site: BamHI (unknown if destroyed)
• 5′ sequencing primer: n/a

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pmCherry-impα was constructed by replacing the AgeI-BglII fragment encoding DsRed of pDsRed-imp α (Addgene plasmid 119679) with mCherry derived from pRSET-B-mCherry (provided by Professor Roger Y. Tsien, UCSD).