pNS19-LV-mutRFP-2A-GFP
(Plasmid #113716)

• Purpose: Lentiviral vector for fluorescent reporter system: expresses a green fluorescent protein that is preceded by an inactive version of a red fluorescent protein (2 ntd substitution at the fluorophore)
• Depositing Lab: Gerald Schwank
• Publication: Savic et al Elife. 2018 May 29;7. pii: 33761. doi: 10.7554/eLife.33761.

Backbone

• Vector backbone: Duet011
• Backbone size w/o insert (bp): 8339
• Total vector size (bp): 11062
• Vector type: Mammalian Expression, Lentiviral ; Reporter system
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: mutRFP-2A-GFP-IRES-Puro
• Alt name: eGFP
• Alt name: mutated turbo RFP
• Alt name: Puromycin resistant gene
• Insert Size (bp): 2723
• Mutation: Y64L (mutRFP)

Resource Information

• Supplemental Documents:
  • pNS19-LV-mutRFP-2A-GFP.gbk

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Cloning of pNS19-LV-mutRFP-2A-GFP: pEGIP (addgene plasmid #26777) was mutagenized using QuikChange Lightning Multi Site-Directed Mutagenesis Kit (Agilent Technologies) to destroy the start codon of eGFP. Next the vector was linearized with BamHI and In-Fusion HD Cloning Plus CE (Takara) was used to insert the mutRFP-2A gBlocks Gene Fragment (Integrated DNA Technologies).