From Tol2Kit
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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. | 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. | ||
Revision as of 01:40, 24 March 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.
Crudely annotated sequence
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:
