pBabe RFP1-Smad2 neo
(Plasmid #58491)

• Purpose: mammalian expression of mRFP1-tagged mouse Smad2
• Depositing Lab: Kevin Janes
• Publication: Wang et al Nat Cell Biol. 2014 Apr;16(4):345-56. doi: 10.1038/ncb2930. Epub 2014 Mar 23.

Backbone

• Vector backbone: pBabe N-terminal RFP neo
• Backbone size w/o insert (bp): 5729
• Modifications to backbone: This construct is a variant of pBabe neo that expresses mRFP1 as an N-terminal fusion. A new XhoI site was inserted immediately upstream of the BamHI site by PCR during cloning. TO EXPRESS AN N-mRFP1 FUSION, GENES MUST HAVE A BAMHI- OR XHO1-COMPATIBLE 5' STICKY END.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Smad2
• Alt name: SMAD family member 2
• Species: M. musculus (mouse)
• Insert Size (bp): 1414
• GenBank ID: NM_010754
• Entrez Gene: Smad2 (a.k.a. 7120426M23Rik, Madh2, Madr2, Smad-2, mMad2)
• Tag / Fusion Protein:
  • mRFP1 (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (destroyed during cloning)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: pBABE 5'
• 3′ sequencing primer: pBABE 3'

Terms and Licenses

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

Depositor Comments

pBabe RFP1_Smad2 neo was constructed by PCR cloning from plasmid templates (Open Biosystems) into the retroviral vector pBabe N-terminal RFP neo. The PCR product was gel purified, digested with BclI and SalI, and then ligated into pBabe N-terminal RFP neo, which had been digested with BamHI and SalI and then dephosphorylated.