LLP185 pLVP-dCas9-DNMT3a V2
(Plasmid #100936)

• Purpose: dCas9 and DNMT WT on C terminus, 4 NLS, driven by pGK promoter, with P2A site and PURO gene, within a lentiviral transfer backbone
• Depositing Lab: Ryan Lister
• Publication: Pflueger et al Genome Res. 2018 Jun 15. pii: gr.233049.117. doi: 10.1101/gr.233049.117.

Backbone

• Vector backbone: pLVP
• Backbone size w/o insert (bp): 6235
• Total vector size (bp): 12934
• Modifications to backbone: Addgene Plasmid #41816 hCas9_D10A with PCR mutagenesis to introduce the H840A mutation to generate the catalytically dead Cas9 (dCas9) and ZF-598-DNMT3A plasmid (Gift from Pilar Blancafort) was used to PCR amplify DNMT3a catalytic domain
• Vector type: Mammalian Expression, Lentiviral, CRISPR
• Selectable markers: Puromycin

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: dCas9, DNMT3a catalytic domain
• Species: H. sapiens (human); S.pyogenes
• Insert Size (bp): 5157
• Mutation: D10A, H840A for dCas9
• Entrez Gene: DNMT3A (a.k.a. DNMT3A2, HESJAS, M.HsaIIIA, TBRS)
• Tags / Fusion Proteins:
  • 3xHA (N terminal on insert)
  • 3xTy1 (C terminal on insert)

Resource Information

• Supplemental Documents:
  • pLVP-dCas9-DNMT WT V2.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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