pCS2-UTX-F-MT2
(Plasmid #40619)

• Purpose: Expression of enzyme-dead UTX in mammalian cells
• Depositing Lab: Kai Ge
• Publication: Wang et al Proc Natl Acad Sci U S A. 2012 Sep 4.

Backbone

• Vector backbone: pCS2+ NLSMT
• Backbone size w/o insert (bp): 4400
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: UTX
• Alt name: ubiquitously transcribed tetratricopeptide repeat, X chromosome
• Species: H. sapiens (human)
• Insert Size (bp): 4200
• Mutation: changed Histidine 1146 and Glutamic Acid 1148 to Alanine
• GenBank ID: NM_021140
• Entrez Gene: KDM6A (a.k.a. KABUK2, UTX, bA386N14.2)
• Promoter: CMV
• Tags / Fusion Proteins:
  • 6xMyc (N terminal on backbone)
  • Flag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Xho I (destroyed during cloning)
• 3′ cloning site: Xba I (not destroyed)
• 5′ sequencing primer: SP6
• 3′ sequencing primer: T7

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

Full length human UTX with a C-terminal FLAG was PCR-amplified from MGC clone BC093868 and cloned into the XhoI-XbaI sites downstream of the CMV promoter and in frame with the N-terminal 6myc tag in the pCS2+ NLSMT vector. Two amino acids essential for UTX enzymatic activity, H1146 and E1148 are mutated to alanine.

UTX insert contains K220E, V503A, N606D, V607A, T726K, Y1173F point mutations relative to closest reference sequence.