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Revision as of 11:52, 12 October 2009 by Chi-bin (talk | contribs) (Construction details)

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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:
PME-mCherryCAAX.png