Ratiometric matryoshka biosensor for Fe2+

MDtxRGA G149 protein

Organism: synthetic construct

Monomeric molecular weight: 84.2 kDa

Oligomeric state: Monomer

Total molecular weight: 84.2 kDa

Sequence:

MRGSHHHHHHHHHHGMASMTGGQQMGRDLYDDDDKDPGRMKDLVDTTEMYLRTIYELEEEGIVPLRARIAERLEQSGPTVSQTVARMERDGLVHVSPDRSLEMTPEGRSLAIAVMRKHRLAERLLTDIIGLDIHKVHDEACRWEHVMSDEVERRLVEVLDDVHRSPFGNPIPGLGEIGLDQADEPDSGPASHNVYITADKQKNGIKANFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTKLSKDPNEKRDHMVLLEFVTAAGITHGMDELYGGTVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAFQTAKLKVTKGGPLPFTWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMIFEDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGLEACEERMYPEDGALKSEYKEWLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERAEGRHSTGGMDELYKGGSASQGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGKYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNNPVRAIDLPLGENLKARIVQLNEILQVDLEQFQALTDAGVEIGTEVDIINEQGRVVITHNGSSVELIDDLAHAVRVEKVEG

The sensor contains different domains: - DtxR (ferrous iron binding protein) from Corynebacterium glutamicum - superfolder circularly permutated GFP (cpsfGFP (reporter fluorophore) from Aequorea victoria - LSSmApple (reference fluorophore) from Discosoma sp. and is constructed as: 1-39 N-Terminal 10xHis, T7tag, enterokinase site/XpressTag, 40-188 DtxR-N-terminus 189-192 PASH Linker 193-281 cpsfGFP-C-terminus 282-284 GGT Linker 285-519 LSSmApple 520-522 GGS Linker 523-666 cpsfGFP-N-terminus 667-670 FNNP Linker 671-749 DtxR-C-terminus