From Tol2Kit
(New page: '''Construction details'''<br> p3E-IRES-EGFPpA was made by fusion PCR of the EMCV IRES sequence, and EGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of ...) |
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'''Construction details'''<br> | '''Construction details'''<br> | ||
| − | p3E-IRES-EGFPpA was made by fusion PCR of the EMCV IRES sequence, and EGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the insert are correct. | + | p3E-IRES-EGFPpA was made by fusion PCR of the EMCV IRES sequence, and EGFP-SV40 late polyA, followed by a BP reaction. 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. Sequencing confirms that all bases of the insert are correct. |
'''Crudely annotated sequence'''<br> | '''Crudely annotated sequence'''<br> | ||
Revision as of 19:31, 7 February 2007
Construction details
p3E-IRES-EGFPpA was made by fusion PCR of the EMCV IRES sequence, and EGFP-SV40 late polyA, followed by a BP reaction. 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. Sequencing confirms that all bases of the insert are correct.
Crudely annotated sequence
FASTA file with the full-length sequence as well as sequences of individual components):
p3E-IRES-EGFPpA sequence
Crude map
Screenshot from Sequencher showing locations of components:
