pGEX-4T-1 3xHA-MKK4-EE
(Plasmid #47577)

• Purpose: Bacterial expression plasmid for GST-3xHA-tagged constitutively active MKK4
• Depositing Lab: Kevin Janes
• Publication: Bose et al Mol Cell Proteomics. 2013 Mar;12(3):797-806. doi: 10.1074/mcp.O112.024059. Epub 2012 Dec 11.

Backbone

• Vector backbone: pGEX-4T-1
• Backbone manufacturer: Amersham
• Backbone size w/o insert (bp): 5062
• Total vector size (bp): 6154
• Modifications to backbone: 3xHA tag inserted at BamHI site of pGEX-4T-1, regenerating the BamHI site downstream of 3xFlag, with a unique HindIII site added between the BamHI and EcoRI sites of pGEX-4T-1.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): BL21
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: MKK4-EE
• Alt name: MAP2K4
• Alt name: MEK4
• Species: H. sapiens (human)
• Insert Size (bp): 1092
• Mutation: Lacks the first 36 residues of the RefSeq protein. Ser257 to Glu, Thr261 to Glu
• GenBank ID: NM_003010
• Entrez Gene: MAP2K4 (a.k.a. JNKK, JNKK1, MAPKK4, MEK4, MKK4, PRKMK4, SAPKK-1, SAPKK1, SEK1, SERK1, SKK1)
• Tags / Fusion Proteins:
  • GST (N terminal on backbone)
  • 3xHA (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: pGEX 5'
• 3′ sequencing primer: pGEX 3'

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Addgene plasmid #14813.

Terms and Licenses

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