Construction and sequencing of a full-length human Scrib cDNA
The KIAA0147 partial cDNA clone was kindly provided by Takahiro Nagase (Kazusa DNA Research Institute, Japan). In order to obtain full-length 5'-cDNA sequences encoding human Scrib, RNA-linker mediated 5'-RACE (RLM-RACE) was performed according to published protocols [49] using RNA isolated from HEK293 cells. The RLM-anchor primer sequence is: 5'-GGGCATAGGCTGACCCTCGCTGAAA-3'. The gene-specific primers are 1) 5'-CACGTCCAGCTCCACCAGCTGCATG-3' and 2) 5'-GAAGTTGGCCACCTCGGGAGGCAAC-3' (nested). This allowed us to construct a composite cDNA of about 5.1 kb which was completely sequenced (Genbank accession no. AF240677).