Reporter

Part:BBa_K1833003:Design

Designed by: Konstantin Borisov   Group: iGEM15_Pitt   (2015-09-16)


pProt-GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 106
    Illegal AgeI site found at 229
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was designed to be used in a cell-free system. As such, we desired the reporter to be transcribed and translated quickly. The promoter was chosen to be a strong sigma-70 E. coli promoter, with the binding sites of the repressor added between the -35 and -10 consensus sequence. Then, the RBS was optimized using the Salis Lab Ribosome Binding Site calculator (https://salislab.net/software/). The combination of the two was used to increase efficiency of in vitro transcription and translation.


Source

This part was synthesized by IDT in two parts, which were then cloned together and into pSB1C3 using Gibson assembly.

References