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Team:TU Delft/Project/rbs-characterization/parts
From 2010.igem.org
BioBricks, the making of
For our RBS characterization project, five different RBS sequences from the Anderson RBS family (J61100, J61101, J61107, J61117, J61127) and the standard RBS B0032 were placed in front of the standard GFP coding sequence. The Biobricks generated in order to perform the experiments were: K398500, K398501, K398502, K398503, K398504. The general map of the construction is shown below, where the RBS is displayed in fucsia.
| Feature | Function |
| AmpR | Ampicillin resistance |
| B0015 | Transcriptional (double) terminator |
| B0062 | Transcriptional terminator |
| E0040 | GFP |
| G00000 | Standard prefix |
| G00001 | Standard suffix |
| G00100 | VF2 primer binding site |
| G00101 | VR primer binding site |
| J61100 | RBS Anderson family |
| J23100 | Promoter |
BBa_I13401 was PCR amplified using the universal primers G00100 and G00101. The purified PCR product was assembled into one of the Anderson RBS plasmids provided in the distribution plates by means of 2-way ligations.
Transformation into competent Top10 E.coli cells yielded positives, as determined by fluorescence analysis.
