pcDNA3-Flag-Tet1 CD Mut
(Plasmid #72218)

• Purpose: Express flag-tagged mouse Tet1 catalytic domain - catalytic dead mutant.
• Depositing Lab: Yi Zhang
• Publication: Ito et al Science. 2011 Sep 2;333(6047):1300-3. doi: 10.1126/science.1210597. Epub 2011 Jul 21.

Backbone

• Vector backbone: pcDNA3
• Backbone size w/o insert (bp): 6000
• Total vector size (bp): 8000
• Modifications to backbone: A rabbit beta globin intron is inserted.
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ten-Eleven Translocation 1
• Species: M. musculus (mouse)
• Mutation: The plasmid expresses TET1 catalytic domain (catalytic dead mutations H1652Y and D1654A).
• Entrez Gene: Tet1 (a.k.a. 2510010B09Rik, Cxxc6, D10Ertd17e, LCX, mKIAA1676)
• Promoter: CMV
• Tag / Fusion Protein:
  • flag (N terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Terms and Licenses

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

Depositor Comments

Modified pcDNA3 vector with a beta globulin intron inserted. This insertion can help stabilize the transcripts and is particularly useful for the expression of large genes, such as p300 and Tet proteins.

Note that an additional mutation, K1581H, was introduced by PCR. This additional mutation does not affect the property of the mutant protein (as it is catalytically dead).

For additional information about how this plasmid was created and used, please see Ito et al., Nature 2010 Pubmed ID: 20639862