Subject: Re: CaSpeR-hs
Dear Lynn,
Here is a sequence run off a GMR vector. The specific insert, a Src29A
cDNA, was cloned after the RI site and is deleted here, so this sequence
should be common to all GMRs. The primer was made to Casper-hs sequence
upstream of the Xho site, pointing forward into the promoter. The sequence
begins with a match to casper-hs up to the Xho site, about base 20. Then
the glass pentamer begins after the Xho- all 5 repeats are perfect. This
goes into a Sal site and polylinker (about base 200), and then the
promoter sequence. It ends with the PCR primer B. Hay lists in
Dev. 120:2121 , terminating in the EcoRI cloning site. Presumably the rest
of GMR is identical to casper-hs. The sequence is SS but unambiguous. Hope
it helps! If you have any trouble putting the whole plasmid together based
on this let me know.
GMR-Src29A promoter region, 497 bp:
TTTGGCAGAAAGAAAAcTCGAGCCCAGTGGAAACCCTTGAAATGCCTTTAAGTCGAG
CCCAGTGGAAACCCTTGAAATGCCTTTAAGTCGAGCCCAGTGGAAACCCTTGAAATG
CCTTTAAGTCGAGCCCAGTGGAAACCCTTGAAATGCCTTTAAGTCGAGCCCAGTGGA
AACCCTTGAAATGCCTTTAAGTCGACTCTAGAGGATCCGCGGCCGCGGGTACCGGGG
GATCCGGCGAAAAGAGCGCCGGAGTATAAATAGAGGCGCTTCGTCTACGGAGCGACA
ATTCAATTCAAACAAGCAAAGTGAACACGTCGCTAAGCGAAAGCTAAGCAAATAAAC
AAGCGCAGCTGAACAAGCTAAACAATCTGCAGTAAAGTGCAAGTTAAAGTGAATCAA
TTAAAAGTAACCAGCAACCAAGTAAATCAACTGCAACTACTGAAATCTGCCAAGAAG
TAATTATTGAATACAAGAAGAGAACTCTGAATAGGGAATTC
Kevin Edwards
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