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URL: http://www.addgene.org/25422
Proper Citation: RRID:Addgene_25422
Insert Name: Myo D 5' flanking sequence
Organism: Homo sapiens
Bacterial Resistance: Ampicillin
Defining Citation: PMID:11458388
Vector Backbone Description: Backbone Marker:Fire et al., 1990; Backbone Size:6110; Vector Backbone:pPD46.21; Vector Types:Mammalian Expression, Reporter construct; Bacterial Resistance:Ampicillin
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).
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Source: Addgene (RRID:SCR_002037)
Description: Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
URL: http://www.addgene.org
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