-24lacZ
(Plasmid #25422)

• Depositing Lab: David J. Goldhamer
• Publication: Chen et al Dev Dyn. 2001 Jul . 221(3):274-88.

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

Terms and Licenses

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

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).