From Tol2Kit
(New page: === Construction details === p3E-mCherrypA was made by PCR amplification of mCherry and an SV40 late polyadenylation signal from a pCS2+-based construct, using primers to add att sites, fo...) |
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=== Construction details === | === Construction details === | ||
p3E-mCherrypA was made by PCR amplification of mCherry and an SV40 late polyadenylation signal from a pCS2+-based construct, using primers to add att sites, followed by a BP reaction. Sequencing finds a single base PCR error, encoding an E7D mutation in mCherry. Functional tests of C-terminal fusion proteins made with this construct show that they yield fluorescent protein. | p3E-mCherrypA was made by PCR amplification of mCherry and an SV40 late polyadenylation signal from a pCS2+-based construct, using primers to add att sites, followed by a BP reaction. Sequencing finds a single base PCR error, encoding an E7D mutation in mCherry. Functional tests of C-terminal fusion proteins made with this construct show that they yield fluorescent protein. | ||
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| + | The mCherry sequence includes a start codon but '''not''' a good Kozak consensus (unlike p3E-EGFP-pA, which has both). | ||
=== Crudely annotated sequence === | === Crudely annotated sequence === | ||
Revision as of 17:04, 30 April 2007
Construction details
p3E-mCherrypA was made by PCR amplification of mCherry and an SV40 late polyadenylation signal from a pCS2+-based construct, using primers to add att sites, followed by a BP reaction. Sequencing finds a single base PCR error, encoding an E7D mutation in mCherry. Functional tests of C-terminal fusion proteins made with this construct show that they yield fluorescent protein.
The mCherry sequence includes a start codon but not a good Kozak consensus (unlike p3E-EGFP-pA, which has both).
Crudely annotated sequence
FASTA file with the full-length sequence as well as sequences of individual components:
p3E-mCherrypA sequence
Crude map
Screenshot from Sequencher showing locations of components:
