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=== Construction details === | === Construction details === | ||
pME-mCherryCAAX was made by PCR amplification of mCherry-CAAX, using primers to add att sites, followed by a BP reaction. mCherry-CAAX is mCherry with a C-terminal fusion of 21 amino acids of human Harvey Ras, encoding a prenylation signal. The original maxiprep that was distributed (clone 232) had an H80D point mutation. Nov 2007: The new version (clone 450) has had this mutation fixed and has no remaining mutations (thanks a lot to Seok-yong Choi). | pME-mCherryCAAX was made by PCR amplification of mCherry-CAAX, using primers to add att sites, followed by a BP reaction. mCherry-CAAX is mCherry with a C-terminal fusion of 21 amino acids of human Harvey Ras, encoding a prenylation signal. The original maxiprep that was distributed (clone 232) had an H80D point mutation. Nov 2007: The new version (clone 450) has had this mutation fixed and has no remaining mutations (thanks a lot to Seok-yong Choi). | ||
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| + | Aug 2008: To our dismay, we realized that our current maxiprep of clone 450 again has mutation C238G, encoding the H80D amino acid change. We are currently reisolating the correct clone. | ||
=== Sequence === | === Sequence === | ||
Revision as of 17:44, 23 August 2008
Construction details
pME-mCherryCAAX was made by PCR amplification of mCherry-CAAX, using primers to add att sites, followed by a BP reaction. mCherry-CAAX is mCherry with a C-terminal fusion of 21 amino acids of human Harvey Ras, encoding a prenylation signal. The original maxiprep that was distributed (clone 232) had an H80D point mutation. Nov 2007: The new version (clone 450) has had this mutation fixed and has no remaining mutations (thanks a lot to Seok-yong Choi).
Aug 2008: To our dismay, we realized that our current maxiprep of clone 450 again has mutation C238G, encoding the H80D amino acid change. We are currently reisolating the correct clone.
Sequence
Annotated sequence, Genbank format:
pME-mCherryCAAX Genbank
FASTA file with the full-length sequence as well as sequences of individual components:
pME-mCherryCAAX sequence
Crude map
Screenshot from Sequencher showing locations of components:
