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Addgene

LEGO-AP1x6-GM55
(Plasmid #217415)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 217415 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    LeGO-iG
  • Backbone manufacturer
    Boris Fehse (Addgene #27358)
  • Vector type
    Mammalian Expression, Lentiviral, Luciferase

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Luciferase, GFP
  • Alt name
    SV40 PolyA sites, 6x AP-1 sites, Human -55 to +28 minimal CSF2 promoter, Promega pGL3 LUC+, IRES-EGFP
  • Species
    H. sapiens (human); Firefly
  • Promoter 6 AP-1 sites, Human -55 to +28 minimal CSF2 promoter

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site unknown (unknown if destroyed)
  • 3′ cloning site unknown (unknown if destroyed)
  • 5′ sequencing primer n/a
  • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This is an inducible lentiviral luciferase expression vector that has 6 RAS/MAPK-inducible AP-1 sites driving transcription from the minimal CSF2 promoter. LEGO-AP1x6-GM55 was constructed from LEGO-AP1x3-GM55 by duplicating the array of 3 AP-1 sites. LEGO-AP1x3-GM55 was previously constructed from LeGO-iG (Weber et al, 2008) (Addgene #27358) by replacing a NotI/XbaI fragment containing the U6 and SFFV promoters with a fragment from the plasmid pAP1-3 (Addgene #71258) that has 3 copies of the MMP3 gene AP-1 site in front of human CSF2 -55 to +28 minimal promoter driving expression of the Luc+ firefly luciferase gene originating from the plasmid pGL3 (Promega). The luciferase gene is linked to an IRES that in principle supports co-expression of luciferase and EGFP. The insert has 2 copies of the SV40 transcription termination/polyadenylation signal upstream of the multiple cloning site, which blocks background read-through transcription originating in the vector backbone, which itself can be substantial (Bert et al, 2000). The inserted luciferase gene and SV40 polyA site sequences originate from the plasmid pXPG (Bert et al, 2000) (Addgene #71248).
References:
Bert et al Plasmid. 2000 Sep;44(2):173-82. PMID 10964627.
Weber et al Mol Ther. 2008 Apr. 16(4):698-706. PMID 18362927.

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:

    LEGO-AP1x6-GM55 was a gift from Peter Cockerill (Addgene plasmid # 217415 ; http://n2t.net/addgene:217415 ; RRID:Addgene_217415)