-24GFP
(Plasmid #25423)

• Depositing Lab: David J. Goldhamer
• Publication: Yamamoto et al Mech Dev. . 124(9-10):715-28.

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

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

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.