piRFP
(Plasmid #31857)

• Purpose: a phytochrome-based near-infrared fluorescent protein, also known as iRFP713, used for in vivo imaging
• Depositing Lab: Vladislav Verkhusha
• Publication: Filonov et al Nat Biotechnol. 2011 Jul 17. doi: 10.1038/nbt.1918.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• 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: Low Copy

Gene/Insert

• Gene/Insert name: iRFP
• Alt name: iRFP713
• Species: Rhodopseudomonas palustris
• Insert Size (bp): 948
• Mutation: T2A, S13L, A92T, V104I, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T and N290Y relative to RpBphP2
• GenBank ID: JN247409
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (destroyed during cloning)
• 3′ cloning site: KpnI (destroyed during cloning)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: SV40pA-R

Resource Information

• Supplemental Documents:
  • Verkhusha lab feature article
  • Verkhusha Lab Spectral data-excel format
• Articles Citing this Plasmid:
  • 36 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

piRFP was derived from pEGFP-C1, whereby iRFP was inserted in place of EGFP. The iRFP in this plasmid has a stop codon at its end making it unsuitable for generating fusions. This plasmid can be used for cytoplasmic expression of iRFP or as a source of the iRFP gene.

For additional protocols on using iRFP in other tissue types please review:
Verkhusha. Deep-tissue photoacoustic tomography of a genetically encoded near-infrared fluorescent probe. Angewandte Chemie Int. Ed. 2012, 51: 1448-1451.
PubMed: https://www.ncbi.nlm.nih.gov/pubmed/22213541/

For more information on in vivo imaging, please see:
https://www.addgene.org/fluorescent_proteins/in_vivo/

Information for Cloning Grade DNA (Catalog # 31857-DNA.cg) ( Back to top )

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here