pmOrange 2-Actin
(Plasmid #199703)

• Purpose: visualization of cleavage furrow in living mammalian mitotic cells
• Depositing Lab: Kenji Sugimoto
• Publication: Tanaka et al Bioimages 25: 7-19, 2017

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Takara Bio.
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 5800
• Modifications to backbone: The pmOrange-actin plasmid was constructed by inserting an mOrange2 DNA fragment, amplified with pcDNA3-mOrange2 (a gift from Dr. Tsien, University of California at San Diego), into the NheI-BglII site and a 1.1-kb DNA fragment of actin cDNA into the BglII site of pEGFP-C1 (Clontech, Palo Alto, CA, USA).
• 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: Unknown

Gene/Insert

• Gene/Insert name: human cytoplasmic action
• Species: H. sapiens (human)
• Insert Size (bp): 700
• Promoter: CMV
• Tag / Fusion Protein:
  • mOrange 2 (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI for mOrange 2, BglII for Actin (unknown if destroyed)
• 3′ cloning site: BglII for mOrange 3, BamH1 for Actin (unknown if destroyed)
• 5′ sequencing primer: pCMV
• 3′ sequencing primer: SV40 Poly A

Resource Information

• A portion of this plasmid was derived from a plasmid made by: pAcGFP-Actin from Takara Bio. pcDNA3-mOrange2 (a gift from Dr. Tsien)

Terms and Licenses

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

Depositor Comments

Nhe I-Bgl II gragment of pAcGFP-Actin (not yet published ) was exchanged with Nhe I-Bgl II 700bp PCR fragment of mOrange2 DNA fragment, amplified with pcDNA3-mOrange2 (a gift from Dr. Tsien, University of California at San Diego).