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Updated: Aug 14, 2026

Massively Parallel Reporter Assays in Cultured Mammalian Cells
Published on: August 17, 2014
Molecular organization of the human Raf-1 promoter region
T W Beck1, U Brennscheidt, G Sithanandam
1Program Resources, Inc., Biological Carcinogenesis Development Program, Frederick, Maryland 21701.
Abstract:
A genomic DNA fragment containing the Raf-1 promoter region was isolated by using a cDNA extension clone. Nucleotide sequencing of genomic DNA clones, primer extension, and S1 nuclease assays have been used to identify the 5' ends of Raf-1 RNAs. Consistent with its ubiquitous expression, the Raf-1 promoter region had features of a housekeeping gene in that it was GC-rich (HTF-like), lacked TATA and CAAT boxes, and contained heterogeneous RNA start sites and four potential binding sites for the transcription factor SP1. In addition, an octamer motif (ATTTCAT), a potential binding site for the octamer family of transcription factors, was located at -734 base pairs. The Raf-1 promoter region drove reporter gene expression 30-fold over the promoterless reporter in Cos 7 cells.
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