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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:
P3E-IRES-EGFPpA.png