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pLenti-Tet-MKK6(DD)-Puro
(Plasmid #86094)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 86094 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pLenti CMV/TO Puro DEST (670-1)
  • Backbone manufacturer
    Eric Campeau
  • Backbone size w/o insert (bp) 8000
  • Total vector size (bp) 9194
  • Modifications to backbone
    L/R recombination to clone in MKK6-DD insert.
  • Vector type
    Mammalian Expression, Lentiviral
  • Selectable markers
    Puromycin

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    MKK6
  • Alt name
    MAP2K6
  • Alt name
    MEK6
  • Alt name
    MKK6-DD, constitutively active mutant
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1005
  • Mutation
    Serine 207 and Threonine 211 both changed to Aspartic Acid (D)
  • GenBank ID
    NM_002758.3
  • Entrez Gene
    MAP2K6 (a.k.a. MAPKK6, MEK6, MKK6, PRKMK6, SAPKK-3, SAPKK3)
  • Promoter CMV/TetO
  • Tag / Fusion Protein
    • Myc tag (N terminal on insert)

Cloning Information

  • Cloning method Gateway Cloning
  • 5′ sequencing primer TGGGCGGTAGGCGTGTACG
  • 3′ sequencing primer GCAGCGTATCCACATAGCG
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    MKK6-DD gene construct originally made in the lab of Angel Nebreda. Citation in comments.
  • Article Citing this Plasmid

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Note: The MKK6 insert in this plasmid has a S2A mutation compared to wild type, but the depositor has confirmed that it does not affect plasmid function.

For proper use, cell line must first be engineered to express the Tetracycline Repressor (TetR). For example: Addgene plasmid #17492. (Not compatible with rtTA system)

Source of MKK6-DD mutant construct:

Alonso, G., Ambrosino, C., Jones, M. and Nebreda, A.R., 2000. Differential activation of p38 mitogen-activated protein kinase isoforms depending on signal strength. Journal of Biological Chemistry, 275(51), pp.40641-40648.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pLenti-Tet-MKK6(DD)-Puro was a gift from Cindy Miranti (Addgene plasmid # 86094 ; http://n2t.net/addgene:86094 ; RRID:Addgene_86094)
  • For your References section:

    Human prostate luminal cell differentiation requires NOTCH3 induction by p38-MAPK and MYC. Frank SB, Berger PL, Ljungman M, Miranti CK. J Cell Sci. 2017 Apr 26. pii: jcs.197152. doi: 10.1242/jcs.197152. 10.1242/jcs.197152 PubMed 28446540