From Tol2Kit
(New page: === Construction details === pME-EGFP was made by PCR amplification of EGFP, using primers that add att sites, followed by a BP reaction. Sequencing confirms that all bases of the insert ...) |
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=== Construction details === | === Construction details === | ||
| − | pME-EGFP was made by PCR amplification of EGFP, using primers that add att sites, followed by a BP reaction. | + | pME-EGFP was made by PCR amplification of the EGFP insert from a pCS2-based plasmid, using primers that add att sites, followed by a BP reaction. Sequencing confirms that all bases of the insert are correct. |
| − | Sequencing confirms that all bases of the insert are correct. | + | |
| + | Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein. | ||
| + | |||
| + | === Sequence === | ||
| + | Annotated sequence, Genbank format:<br> | ||
| + | [[pME-EGFP 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-EGFP sequence]] | [[pME-EGFP sequence]] | ||
Latest revision as of 03:44, 19 December 2007
Construction details
pME-EGFP was made by PCR amplification of the EGFP insert from a pCS2-based plasmid, using primers that add att sites, followed by a BP reaction. Sequencing confirms that all bases of the insert are correct.
Note that the EGFP sequence is followed by 60 bp of polylinker sequence from pCS2+ before the stop codon. We have confirmed functionally that this cassette produces fluorescent protein.
Sequence
Annotated sequence, Genbank format:
pME-EGFP Genbank
FASTA file with the full-length sequence as well as sequences of individual components:
pME-EGFP sequence
Crude map
Screenshot from Sequencher showing locations of components:
