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Addgene

-24GFP
(Plasmid #25423)

Full plasmid sequence is not available for this item.

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 25423 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pEGFP-1
  • Backbone size w/o insert (bp) 4200
  • Vector type
    Mammalian Expression ; Reporter construct

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Myo D 5' flanking sequence
  • Alt name
    MyoD
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    24000
  • Entrez Gene
    MYOD1 (a.k.a. CMYP17, MYF3, MYOD, MYODRIF, PUM, bHLHc1)
  • Tag / Fusion Protein
    • EGFP (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SalI (not destroyed)
  • 3′ cloning site AscI (not destroyed)
  • 5′ sequencing primer N/A
  • 3′ sequencing primer EGFP-N
  • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

The MyoDGFP (-24GFP) transgene was constructed by excising 24 kb of human MyoD 5' flanking sequence from -24lacZ (Chen et al., 2001; Addgene plasmid #25422) by digestion with SalI and AscI (AscI linkers were added to the unique XhoI site at the 3' end of human genomic sequences) and inserting into SalI and AscI (produced by linker addition to the unique SmaI site) sites in the polylinker of pEGFP-1 (Clontech). Prior to subcloning, the NotI site just 3' of EGFP was destroyed by blunt-ending with Klenow, and a new NotI site was introduced by linker addition to the AflII site, which lies 3' of SV40 poly sequences.

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:

    -24GFP was a gift from David J. Goldhamer (Addgene plasmid # 25423 ; http://n2t.net/addgene:25423 ; RRID:Addgene_25423)
  • For your References section:

    Cloning and characterization of a novel MyoD enhancer-binding factor. Yamamoto M, Watt CD, Schmidt RJ, Kuscuoglu U, Miesfeld RL, Goldhamer DJ. Mech Dev. . 124(9-10):715-28. 10.1016/j.mod.2007.07.002 PubMed 17693064