pLX304-EZH1-D745N
(Plasmid #203585)

• Purpose: Expresses V5-tagged mutant version of EZH1 partially resistant to JQ-EZ-05 in mammalian cells.
• Depositing Labs: Abhishek Chakraborty, William Kaelin
• Publication: Chakraborty et al Sci Transl Med. 2017 Jul 12;9(398):eaal5272. doi: 10.1126/scitranslmed.aal5272.

Backbone

• Vector backbone: pLX304
• Backbone size w/o insert (bp): 9377
• Total vector size (bp): 9963
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Blasticidin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: EZH1
• Alt name: Enhancer of zest 1 Polycomb repressive complex 2
• Alt name: KMT6B
• Species: H. sapiens (human)
• Insert Size (bp): 2313
• Mutation: changed Aspartic Acid 745 to Asparagine for partial resistance to JQ-EZ-05
• GenBank ID: 2145
• Entrez Gene: EZH1 (a.k.a. KMT6B)
• Promoter: CMV
• Tag / Fusion Protein:
  • V5-tagged (C terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: WPRE-R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Broad TRC (ccsbBroad304_00525)

Terms and Licenses

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

Depositor Comments

Mutations encoding the D745F were introduced by site-directed mutagenesis into the pLX304-EZH1 backbone