Part:BBa_K4218002

This part is composite of T7 promoter, double toehold switch part BBa_K3822000,FP mcherry and termin

We have established a toehold switch that combines with a fluorescent protein, mCherry, on its tail to detect methylated TFPI2 for early screening of non-invasive diagnosis of CRC (https://2021.igem.org/Team:YiYe-China). It is reported that SDC2 methylation and TFPI2 methylation are highly sensitive to CRC (DOI:10.2147/cmar.s375358) . In order to further improve the detection accuracy of CRC compared to last year, we combined the detection of methylated SDC2 with methylated TFPI2 this year. We constructed a toehold switch consisting of the plasmids of double-toehold pSB1C3 (more details were listed below) and double-trigger pCOLADuetTM-1, and then we have a functional verification for the plasmids in the BL21 cells. Finally, we detected methylation SDC2 gene and methylated TFPI2 through the expression of fluorescent protein in cell-free protein expression system.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]