Designed by: Dong-Jiunn Jeffery TRUONG Group: iGEM13_TU-Munich (2013-09-16)
FRET Reporter for TEV Protease activity (eCFP_TEV-site-linker_SYFP2) in RFC
This part encodes a FRET reporter with the chromophores eCFP and SYFP2 that is seperated by a TEV cleavage site. As a FRET pair, excitation of eCFP leads to a energy transfer from eCFP to SYFP2 (FRET) resulting in a fluorescence of SYFP2. In the presence of TEV Protease cleavage actitivy the FRET pair SYFP2 is cleaved off from eCFP and eCFP is not anymore quenched by SYFP2 resulting in the fluorescence of eCFP. This part is flanked by RFC pre- and suffix for further protein fusions.
Nucleotide sequence in RFC 25, so ATGGCCGGC and ACCGGT were added (in italics) to the 5' and 3' ends: (underlined part encodes the protein) ATGGCCGGCGTGAGCAAG ... CTGTATAAAACCGGT ORF from nucleotide position -8 to 1473 (excluding stop-codon)
Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)