pET28b-MBP-TEV-PWWP_DNMT3A
(Plasmid #186970)

• Purpose: Bacterial expression of human DNMT3A PWWP domain (residues 278–427) with His-MBP tag and TEV protease cleavage site.
• Depositing Lab: Brian Liau
• Publication: Lue et al Nat Chem Biol. 2023 Feb;19(2):176-186. doi: 10.1038/s41589-022-01167-4. Epub 2022 Oct 20.

Backbone

• Vector backbone: pET28b
• Backbone size w/o insert (bp): 5222
• Total vector size (bp): 6932
• 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: DNMT3A PWWP domain
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 1750
• Mutation: PWWP domain (residues 278–427)
• GenBank ID: NM_022552.4
• Entrez Gene: DNMT3A (a.k.a. DNMT3A2, HESJAS, M.HsaIIIA, TBRS)
• Promoter: T7
• Tag / Fusion Protein:
  • His-MBP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pET-SF CAGCAACCGCACCTGTGG
• 3′ sequencing primer: T7-SR GCTAGTTATTGCTCAGCGG

Resource Information

• Supplemental Documents:
  • pET28b-MBP-TEV-PWWP_DNMT3A.gb
• A portion of this plasmid was derived from a plasmid made by: The His-MBP-TEV insert was derived from MBP-FUS_FL_WT, a gift from N. Fawzi (Addgene #98651).

Terms and Licenses

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