1GFP/RNase H1 D210N
(Plasmid #174448)

• Purpose: Express GFP-tagged RNase H1 D210N in E. coli.
• Depositing Lab: Karlene Cimprich
• Publication: Crossley et al J Cell Biol. 2021 Sep 6;220(9). pii: 212458. doi: 10.1083/jcb.202101092. Epub 2021 Jul 7.

Backbone

• Vector backbone: 1GFP
• Backbone manufacturer: California Institute for Quantitative Biosciences, University of California, Berkeley MacroLab
• Backbone size w/o insert (bp): 6075
• Total vector size (bp): 6858
• Vector type: Bacterial Expression

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: RNase H1
• Species: H. sapiens (human)
• Insert Size (bp): 783
• Mutation: D210N
• Entrez Gene: RNASEH1 (a.k.a. H1RNA, PEOB2, RNH1)
• Promoter: T7 promoter
• 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

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

Expression of His-GFP-RNase H1 D210N was induced with 0.3 mM IPTG at 16C for 18-20 hours in BL21(DE3)/Rosetta after OD600 reaching 0.3-0.5.