pPL6873_2xcHS4::mCherry::2xcHS4_FRT-PuroR-P2A-mTagBFP2-NLS
(Plasmid #217970)

• Purpose: Provides expression from a CMV promoter surrounded by insulators. Integrates at FRT site. Confers puromycin resistance. Expresses nuclear localized BFP.
• Depositing Lab: Robert Piper
• Publication: Piper Lab Plasmids 2024 (unpublished)

Backbone

• Vector backbone: pcDNA5
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 2xcHS4::mCherry::2xcHS4_FRT-PuroR-P2A-mTagBFP2-NLS
• Species: Synthetic

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• Supplemental Documents:
  • pPL6873_2xcHS4--mCherry--2xcHS4_FRT-PuroR-P2A-mTagBFP2-NLS.gbk

Terms and Licenses

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

Depositor Comments

This plasmid integrates at FRT sites when co-transfected with a plasmid producing Flp-recombinase. This plasmid has two ORFS. The first ORF is built to take advantage of the 'Flp-in' system whereby a genomic FRT site provides an ATG translational start site for the puromycin resistance gene and nuclear-localized blue fluorescent protein. These two proteins are separated by a P2A cleavage site. The second ORF is surrounded by tandem core cHS4 insulators. The insulated ORF encodes the CMV promoter driving the production of mCherry. mCherry can be released by restriction enzymes and replaced with a gene of interest.