pmCherry-C1 R62D actin-3XNLS P2A mCherry
(Plasmid #58477)

• Purpose: Expresses nuclear-targeted non-polymerizing R62D mutant of human actin, with an mCherry expression reporter after a P2A protease cleavage site, on a CMV promoter
• Depositing Lab: Dyche Mullins
• Publication: Belin et al Elife. 2015 Aug 19;4. doi: 10.7554/eLife.07735.

Backbone

• Vector backbone: mCherry-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 5994
• 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): Top10
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: beta actin
• Species: H. sapiens (human)
• Insert Size (bp): 1125
• Mutation: Changed Arginine 62 to Aspartic Acid
• GenBank ID: NM_001101.3
• Entrez Gene: ACTB (a.k.a. BKRNS, BNS, BRWS1, CSMH, DDS1, PS1TP5BP1, THC8)
• Promoter: CMV

Cloning Information for Gene/Insert 1

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: CMV Forward
• 3′ sequencing primer: mCherry Reverse

Gene/Insert 2

• Gene/Insert name: 3XNLS
• Alt name: nuclear localization signal
• Species: SV40 virus
• Insert Size (bp): 72
• Promoter: CMV

Cloning Information for Gene/Insert 2

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: actin Forward
• 3′ sequencing primer: mCherry Reverse

Gene/Insert 3

• Gene/Insert name: P2A cleavage site
• Alt name: 2A Peptide Derived from Porcine Teschovirus-1
• Species: Porcine Teschovirus-1
• Promoter: CMV
• Tag / Fusion Protein:
  • mCherry (C terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: actin Forward
• 3′ sequencing primer: mCherry Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: P2A peptide and 3X NLS (with stop codons) were inserted into the pmCherry-C1 backbone via GenScript's custom cloning services. Human actin was cloned from cDNA using Gibson cloning into an NLS-P2A-mCherry backbone.
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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