Part:BBa_K1859011

2*GGSGGS linker+ The 5´side of the intron [BBa_K1332003]

[BBa_K1859011] is the 5'side of the intron [BBa_K1332003] conjuncted linker to keep folding of proteins. Circular Parts, the 3'side of the intron [BBa_K1332005] and the 5'side of the intron [BBa_K1332003] , is what use sequence subcloned ribozyme involved self‐splicing in T4 Phage. By associating Circular Parts on both ends of optional sequence, you can circularize mRNA making its DNA.

As for details, refer to iGEM Gifu 2014

In the study of iGEM Gifu 2014, we made long-chain protein from Circular mRNA, but it lost functionality. It is thought that folding of the protein failed for a cause． Therefore we designed some linkers to fix the folding of the protein. [BBa_K1859011] includes sequence of the 5'side of the intron [BBa_K1332003] and linker． The linker comes inside the Circular mRNA when mRNA is circularized. To make this device, we combined the sequence of the linker, 2*GGSGGS , to the upstream of 5'side of the intron [BBa_K1332003] .

The linker designed elsewhere is here .

We attached this part[BBa_K1859011] to 3'sides of His-RFP without stop codon [BBa_K1332002] . Then, it was connected to 3'sides of [BBa_K1859007] . In addition,Lacl [BBa_R0010] , RBS [BBa_B0034] and DT [BBa_R0015] was conbined with it, making [BBa_K1859020] . [BBa_K1859020] is the generator that express long-chain protein with keeping functionality of the protein.