Part:BBa_I742130:Experience

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Glasgow 2014 experience

We tested this reversed RBS in TOP10 and DH5alpha cells upstream (OK downstream) of our reversed RFP (Red Fluorescent Protein) biobrick BBa_K1463520. This can be seen in Fig. 1 below.

It gave a lower fluorescence than our designed reverse RBS BBa_K1463560.

Fig. 1: Red fluorescence of colonies containing BBa_K1463560 or BBa_I742130 reversed RBSs "downstream" of the reversed rfp gene BBa_K1463520 transcribed from the PhiC31 integrase switch BBa_K1463000. The image was scanned using a GE Healthcare Typhoon FLA9500 using the 532 nm laser and LPG filter.

Consistent with this result, BBa_I742130 had a lower predicted translation initiation rate than BBa_K1463560 as calculated by the Salis RBS calculator.

BBa_I742130 calculated translation initiation rate 495.33

BBa_K1463560 calculated translation initiation rate 12,204.43

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