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 === | |
| − | 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 (IRES2 from Clontech), 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. |
| + | |||
| + | === Sequence === | ||
| + | Annotated sequence, Genbank format:<br> | ||
| + | [[p3E-IRES-EGFPpA 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> | ||
[[p3E-IRES-EGFPpA sequence]] | [[p3E-IRES-EGFPpA sequence]] | ||
| − | + | === Crude map === | |
Screenshot from Sequencher showing locations of components:<br> | Screenshot from Sequencher showing locations of components:<br> | ||
[[Image:p3E-IRES-EGFPpA.png]] | [[Image:p3E-IRES-EGFPpA.png]] | ||
Latest revision as of 18:48, 17 August 2011
Construction details
p3E-IRES-EGFPpA was made by fusion PCR of the EMCV IRES sequence (IRES2 from Clontech), 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.
Sequence
Annotated sequence, Genbank format:
p3E-IRES-EGFPpA Genbank
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:
