From Tol2Kit
(New page: === Construction details === p3E-EGFPpA was made by PCR amplification of EGFP and an SV40 late polyadenylation signal from a pCS2+-based plasmid, using primers to add att sites, followed b...) |
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p3E-EGFPpA was made by PCR amplification of EGFP and an SV40 late polyadenylation signal from a pCS2+-based plasmid, using primers to add att sites, followed by a BP reaction. | p3E-EGFPpA was made by PCR amplification of EGFP and an SV40 late polyadenylation signal from a pCS2+-based plasmid, using primers to add att sites, 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. | + | 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 in C-terminal fusion constructs. Sequencing confirms that all bases of the insert are correct. |
=== Crudely annotated sequence === | === Crudely annotated sequence === | ||
Revision as of 03:07, 24 March 2007
Construction details
p3E-EGFPpA was made by PCR amplification of EGFP and an SV40 late polyadenylation signal from a pCS2+-based plasmid, using primers to add att sites, 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 in C-terminal fusion constructs. 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-EGFPpA sequence
Crude map
Screenshot from Sequencher showing locations of components:
