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.
The EGFP sequence does include a start codon and a good Kozak consensus, so it is possible to use this clone to test promoters cloned into a middle clone.
Sequence
Annotated sequence, Genbank format:
p3E-EGFPpA Genbank
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:
