From Tol2Kit
(New page: '''Construction details'''<br> p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence, and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bas...) |
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| − | + | === Construction details === | |
| − | p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence, and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the coding sequence are correct. There is a single base change in the IRES sequence, but we have confirmed functionally that this cassette produces fluorescent, nuclear-localized protein. | + | p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence (IRES2 from Clontech), and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the coding sequence are correct. There is a single base change in the IRES sequence (G481A), but we have confirmed functionally that this cassette produces fluorescent, nuclear-localized protein. |
| − | + | === Sequence === | |
| − | FASTA file with the full-length sequence as well as sequences of individual components | + | Annotated sequence, Genbank format:<br> |
| + | [[p3E-IRES-nlsEGFPpA Genbank]] | ||
| + | |||
| + | FASTA file with the full-length sequence as well as sequences of individual components:<br> | ||
[[p3E-IRES-nlsEGFPpA sequence]] | [[p3E-IRES-nlsEGFPpA sequence]] | ||
| − | + | === Crude map === | |
Screenshot from Sequencher showing locations of components:<br> | Screenshot from Sequencher showing locations of components:<br> | ||
[[Image:p3E-IRES-nlsEGFPpA.png]] | [[Image:p3E-IRES-nlsEGFPpA.png]] | ||
Latest revision as of 18:47, 17 August 2011
Construction details
p3E-IRES-nlsEGFPpA was made by fusion PCR of the EMCV IRES sequence (IRES2 from Clontech), and nlsEGFP-SV40 late polyA, followed by a BP reaction. Sequencing confirms that all bases of the coding sequence are correct. There is a single base change in the IRES sequence (G481A), but we have confirmed functionally that this cassette produces fluorescent, nuclear-localized protein.
Sequence
Annotated sequence, Genbank format:
p3E-IRES-nlsEGFPpA Genbank
FASTA file with the full-length sequence as well as sequences of individual components:
p3E-IRES-nlsEGFPpA sequence
Crude map
Screenshot from Sequencher showing locations of components:
