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PurposePlasmid for expression of bacterial RNase HI tagged with NLS-mCherry that can be used to degrade R-loops. Confers resistance to Puromycin. Expression is inducible in T-REx cells.
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Depositing Labs
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 60365 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepICE
- Backbone size w/o insert (bp) 5044
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Vector typeMammalian Expression
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Selectable markersPuromycin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)XL1 Blue
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameRNase HI
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Alt namernhA
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Alt namedasF
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Alt nameherA
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SpeciesEscherichia coli
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Insert Size (bp)495
- Promoter CMV-tet
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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 (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pICE-RNaseHI-WT-NLS-mCherry was a gift from Sébastien Britton & Patrick Calsou (Addgene plasmid # 60365 ; http://n2t.net/addgene:60365 ; RRID:Addgene_60365) -
For your References section:
DNA damage triggers SAF-A and RNA biogenesis factors exclusion from chromatin coupled to R-loops removal. Britton S, Dernoncourt E, Delteil C, Froment C, Schiltz O, Salles B, Frit P, Calsou P. Nucleic Acids Res. 2014 Oct 1;42(14):9047-62. doi: 10.1093/nar/gku601. Epub 2014 Jul 16. 10.1093/nar/gku601 PubMed 25030905