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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). | ||
| − | === | + | 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. |
| + | |||
| + | Oct 2009: We reisolated the correct clone, now named clone <em>550</em>, and have been distributing this for quite a while. Clone 450 is now obsolete. | ||
| + | |||
| + | === Sequence === | ||
| + | Annotated sequence, Genbank format:<br> | ||
| + | [[pME-mCherryCAAX Genbank]] | ||
| + | |||
FASTA file with the full-length sequence as well as sequences of individual components:<br> | FASTA file with the full-length sequence as well as sequences of individual components:<br> | ||
[[pME-mCherryCAAX sequence]] | [[pME-mCherryCAAX sequence]] | ||
Latest revision as of 11:55, 12 October 2009
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.
Oct 2009: We reisolated the correct clone, now named clone 550, and have been distributing this for quite a while. Clone 450 is now obsolete.
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:
