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MLRV2: NF-kb-SMAD-DBE-P53-AP1
(Plasmid #124536)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 124536 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    Omega Destination-CMV::ELuc:bGH
  • Backbone manufacturer
    Venken Lab, #124528
  • Backbone size w/o insert (bp) 5000
  • Total vector size (bp) 13636
  • Vector type
    Mammalian Expression, Luciferase, Synthetic Biology

Growth in Bacteria

  • Bacterial Resistance(s)
    Spectinomycin, 50 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert 1

  • Gene/Insert name
    5 copies of the NF-kb DNA binding motif
  • Alt name
    Transcription Blocker + 5 copies of the NF-kb DNA binding motif + miniP+ Red Firefly luciferase
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    2215

Cloning Information for Gene/Insert 1

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsmBI (destroyed during cloning)
  • 3′ cloning site BsmBI (destroyed during cloning)
  • 5′ sequencing primer ACTCCACCCATTGACGTCAATGG
  • 3′ sequencing primer agaatggcgccgggcctttc
    • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    7xSMAD_RE::FLuc
  • Alt name
    Transcription Blocker + 7 copies of the SMAD DNA binding motif + miniP+ Firefly luciferase
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    2276

Cloning Information for Gene/Insert 2

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsmBI (destroyed during cloning)
  • 3′ cloning site BsmBI (destroyed during cloning)
  • 5′ sequencing primer CAAGAAGCCCGTGGCCAAGATG
  • 3′ sequencing primer CGGGGTCATAAACCTTGGAAGTCATTG
    • (Common Sequencing Primers)

Gene/Insert 3

  • Gene/Insert name
    3xDBE_BE::Renilla (FOXO)
  • Alt name
    Transcription Blocker + 3 copies of the DBE DNA binding motif + miniP + Renilla luciferase
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    1543

Cloning Information for Gene/Insert 3

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsmBI (destroyed during cloning)
  • 3′ cloning site BsmBI (destroyed during cloning)
  • 5′ sequencing primer agggcggaaagatcgccgtg
  • 3′ sequencing primer CGAAGTCCTCCAAGGTAAACACCATTG
    • (Common Sequencing Primers)

Gene/Insert 4

  • Gene/Insert name
    2xP53_RE::NLuc
  • Alt name
    Transcription Blocker + 2 copies of the p53 DNA binding motif + miniP + NLuc luciferase
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    1088

Cloning Information for Gene/Insert 4

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsmBI (destroyed during cloning)
  • 3′ cloning site BsmBI (destroyed during cloning)
  • 5′ sequencing primer GTCCTGAAGAACGAACAGTGAGCTTC
  • 3′ sequencing primer CTGGGTCGTAGACTTTGGAAGCC
    • (Common Sequencing Primers)

Gene/Insert 5

  • Gene/Insert name
    6xAP-1_RE::GrRenilla
  • Alt name
    Transcription Blocker + 6 copies of the AP-1 DNA binding motif + miniP + GreenRenilla luciferase
  • Insert Size (bp)
    1514

Cloning Information for Gene/Insert 5

  • Cloning method Restriction Enzyme
  • 5′ cloning site BsmBI (unknown if destroyed)
  • 3′ cloning site BsmBI (unknown if destroyed)
  • 5′ sequencing primer GAGGCTCTGCGAACGAATACTCG
  • 3′ sequencing primer ctt ttt acg gtt cct ggc ctt ttg
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.
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:

    MLRV2: NF-kb-SMAD-DBE-P53-AP1 was a gift from Koen Venken (Addgene plasmid # 124536 ; http://n2t.net/addgene:124536 ; RRID:Addgene_124536)

  • For your References section:

    Rapid and Efficient Synthetic Assembly of Multiplex Luciferase Reporter Plasmids for the Simultaneous Monitoring of Up to Six Cellular Signaling Pathways. Sarrion-Perdigones A, Gonzalez Y, Venken KJT. Curr Protoc Mol Biol. 2020 Jun;131(1):e121. doi: 10.1002/cpmb.121. 10.1002/cpmb.121 PubMed 32539183
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