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Addgene

-24lacZ
(Plasmid #25422)

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 25422 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pPD46.21
  • Backbone manufacturer
    Fire et al., 1990
  • Backbone size w/o insert (bp) 6110
  • Vector type
    Mammalian Expression ; Reporter construct

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 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
    • lacZ (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SalI (unknown if destroyed)
  • 3′ cloning site AscI (unknown if destroyed)
  • 5′ sequencing primer N/A
  • 3′ sequencing primer LacZ-R
  • (Common Sequencing Primers)

Terms and Licenses

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

Depositor Comments

-24lacZ contains approximately 24 kb of 5' flanking sequences of the human MyoD gene, cloned upstream of the lacZ reporter gene in the vector pPD46.21 (Fire et al., 1990).

MyoD flanking sequences were derived from the CAT reporter construct, -24CAT (Goldhamer et al., 1992), as follows. The unique XhoI site, which is just 3' of human sequences in -24CAT, was Klenow-filled and AscI linkers (New England Biolabs) were added by blunt-end ligation using standard methods. AscI linkers were also added to the unique SmaI site in the polylinker of pPD46.21. The 24-kb fragment was excised from -24CAT by digestion with SalI and AscI, purified by agarose gel electrophoresis and GENECLEAN Xbio-gene), and directionally cloned into pPD46.21 that had been similarly prepared.

The resulting clone contains contiguous MyoD 5' flanking sequences from approximately 24 kb upstream to 170 bp downstream of the MyoD transcriptional start site (-37 relative to the start of translation).

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:

    -24lacZ was a gift from David J. Goldhamer (Addgene plasmid # 25422 ; http://n2t.net/addgene:25422 ; RRID:Addgene_25422)
  • For your References section:

    Two upstream enhancers collaborate to regulate the spatial patterning and timing of MyoD transcription during mouse development. Chen JC, Love CM, Goldhamer DJ. Dev Dyn. 2001 Jul . 221(3):274-88. 10.1002/dvdy.1138 PubMed 11458388