pPL7552_ColE1_PiggyBac_2xcHS4::mCherry-IRES-PuroR-P2A-mTagBFP2-NLS::2xcHS4
(Plasmid #217971)

• Purpose: Provides expression from a CMV promoter surrounded by insulators. Integrates via piggyBac transposase. Confers puromycin resistance. Expresses nuclear localized BFP. High copy plasmid.
• Depositing Lab: Robert Piper
• Publication: Piper Lab Plasmids 2024 (unpublished)

Backbone

• Vector backbone: Bluescript
• 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-IRES-PuroR-mTagBFP2-NLS::2xcHS4
• Species: Synthetic

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: n/a

Resource Information

• Supplemental Documents:
  • pPL7552_ColE1_PiggyBac_2xcHS4--mCherry-IRES-PuroR-P2A-mTagBFP2-NLS--2xcHS4.gbk

Terms and Licenses

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

Depositor Comments

This plasmid integrates throughout the genome when co-expressed with piggyBac transposase. This plasmid expresses mCherry, which can be removed by restriction enzymes and replaced with a gene of interest. Expressed from the same mRNA via a downstream Internal Ribosome Entry Site is a fusion protein consisting of the Puromycin-resistance protein, a P2A site, and nuclear-localized blue fluorescent protein. The fusion protein is cleaved at the P2A site, producing two separate proteins. The ORF is expressed via the CMV promoter. The CMV_mCherry_IRES_Puro_P2A_mTagBFP2-NLS insert is flanked by dual core cHS4 insulators that potentiate the expression when stably integrated in the genome. This is a high copy plasmid.