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PurposeExpress GFP-tagged RNase H1 WT in E. coli.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 174447 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbone1GFP
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Backbone manufacturerCalifornia Institute for Quantitative Biosciences, University of California, Berkeley MacroLab
- Backbone size w/o insert (bp) 6075
- Total vector size (bp) 6858
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberLow Copy
Gene/Insert
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Gene/Insert nameRNase H1
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SpeciesH. sapiens (human)
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Insert Size (bp)783
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Entrez GeneRNASEH1 (a.k.a. H1RNA, PEOB2, RNH1)
- Promoter T7 promoter
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Tag
/ Fusion Protein
- His-tag, GFP (N terminal on backbone)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer GFP forward primer: 5'-GTCCTGCTGGAGTTCGTGACC-3'
- 3′ sequencing primer T7 terminator reverse primer (Common Sequencing Primers)
Resource Information
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Article 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.
Depositor Comments
Expression of His-GFP-RNase H1 WT was induced with 0.3 mM IPTG at 16C for 18-20 hours in BL21(DE3)/Rosetta after OD600 reaching 0.3-0.5.
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:
1GFP/RNase H1 WT was a gift from Karlene Cimprich (Addgene plasmid # 174447 ; http://n2t.net/addgene:174447 ; RRID:Addgene_174447) -
For your References section:
Catalytically inactive, purified RNase H1: A specific and sensitive probe for RNA-DNA hybrid imaging. Crossley MP, Brickner JR, Song C, Zar SMT, Maw SS, Chedin F, Tsai MS, Cimprich KA. J Cell Biol. 2021 Sep 6;220(9). pii: 212458. doi: 10.1083/jcb.202101092. Epub 2021 Jul 7. 10.1083/jcb.202101092 PubMed 34232287