RED
(Plasmid #139835)

• Purpose: Expresses RNase H1-EGFP- dCas9 fusion protein in human cells
• Depositing Lab: Karim Mekhail
• Publication: Abraham et al Nature. 2020 Sep;585(7824):298-302. doi: 10.1038/s41586-020-2497-0. Epub 2020 Jul 15.

Backbone

• Vector backbone: pcDNA4/TO
• Backbone manufacturer: Life Technologies/Thermo Fisher
• Backbone size w/o insert (bp): 5078
• Total vector size (bp): 10922
• Vector type: Mammalian Expression
• Selectable markers: Zeocin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: RNase H1-EGFP-dCas9
• Alt name: RNase H1-EGFP-deadCas9
• Species: RNase H1 (H. sapiens), dCas9 (S. pyogenes)
• Insert Size (bp): 5844
• Mutation: dCas9 protein contains two mutations: D10A and H840A
• GenBank ID: RNase H1 (BT019670.1) Cas9 (AAK33936.1)
• Entrez Gene: RNASEH1 (a.k.a. H1RNA, PEOB2, RNH1)
• Promoter: tet-inducible CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: BGH reverse

Resource Information

• Supplemental Documents:
  • Annotated Depositor's Sequence
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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