pCDFDuet-1 MKK4(EE)-MKK7a1(EE)
(Plasmid #47580)

• Purpose: Bacterial expression plasmid for co-expression of constitutively active MKK4 and MKK7a1
• 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: pCDFDuet-1
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 3781
• Total vector size (bp): 5851
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): BL21
• Copy number: Low Copy

Gene/Insert 1

• 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)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SalI (not destroyed)
• 5′ sequencing primer: ACYCDuetUP1
• 3′ sequencing primer: DuetDOWN1

Gene/Insert 2

• Gene/Insert name: MKK7a1-EE
• Alt name: Map2k7
• Alt name: Mek7
• Species: M. musculus (mouse)
• Insert Size (bp): 1041
• Mutation: Residues 73-420 of RefSeq sequence. Ser271 to Glu, Thr275 to Glu.
• GenBank ID: NM_001042557
• Entrez Gene: Map2k7 (a.k.a. 5930412N11Rik, JNKK 2, Jnkk, Jnkk2, MAPKK 7, MEK 7, Mapkk7, Mek7, Mkk7, Prkm, Prkmk7, sek, sek2)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (destroyed during cloning)
• 3′ cloning site: XhoI (destroyed during cloning)
• 5′ sequencing primer: DuetUP2 Primer
• 3′ sequencing primer: T7 Terminator Primer

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Addgene plasmids #47577 and #47578.
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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