pICE-RNaseHI-D10R-E48R-NLS-mCherry
(Plasmid #60367)

• Purpose: Plasmid for expression of an inactive mutant of bacterial RNase HI tagged with NLS-mCherry that can be used to detect R-loops. Confers resistance to Puromycin. Expression is inducible in T-REx cells.
• Depositing Labs: Sébastien Britton, Patrick Calsou
• Publication: Britton et al Nucleic Acids Res. 2014 Oct 1;42(14):9047-62. doi: 10.1093/nar/gku601. Epub 2014 Jul 16.

Backbone

• Vector backbone: pICE
• Backbone size w/o insert (bp): 5044
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: RNase HI-D10R-E48R
• Alt name: rnhA
• Alt name: dasF
• Alt name: herA
• Species: Escherichia coli
• Insert Size (bp): 495
• Mutation: D10R+E48R; catalyticaly inactive
• Promoter: CMV-tet
• Tags / Fusion Proteins:
  • NLS from SV40 T antigen (C terminal on insert)
  • mCherry (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: CMVpromoter forward
• 3′ sequencing primer: BGHpolyA reverse

Resource Information

• Supplemental Documents:
  • Author annotated GenBank sequence
• Articles Citing this Plasmid:
  • 10 References

Terms and Licenses

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