P3E-EGFPpA

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[edit] 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.

[edit] 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

[edit] Crude map

Screenshot from Sequencher showing locations of components:
Image:p3E-EGFPpA.png

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