Part:BBa_K797002
TorA signal with RBS
This part is coding N-terminal torA signal region. Proteins conjugated torA signal are recognized by Tat secretion pathway and they are secreted from cytoplasm to periplasm while maintaining their holdings. This come from the part with the enzym of TMAOreductase. This parts include RBS so that you don't bother to insert new RBS between this signal and your promoter. ( Note: Depending on your purpose, you might use BBa_K797004 when you use this signal because wild type Tat pathway is relatively
saturated easily.)
In this experiment, we wanted to design an applicable TorA signal device to meet various needs and to check the function of signal sequence. TorA signal was, actually,submitted by Canbrige 2011(BBa_K233307) cause a stop codon between signal peptide and target coding sequence(CDS) when you assemble them by standard or 3A assembly. For these reasons, all of other teams make an effort to combine TorA signal to targets, such as using Gibson assembly. That's too trouble!
To avoid appearing a stop codon in scar sequence between TorA signal and a target protein CDS, we made Frameshift mutation twice on TorA signal. As a result, you can make TorA-fusion target protein by standard or 3A assembly. In addition to that, our TorA signal has RBS. This can be an easy-to-use biobrick part. It can shorten the time of constructions because it only needs a promoter and a target protein.
We read the sequence data of our modified TorA signal and confirmed stop codon doesn't appear when it used in Standard or 3A assembly. Using green fluorescent protein (GFP) as a target protein, we observed the TorA-GFP fusion-expressing cells. The TorA-GFP fusion was successfully expressed. This means RBS in our TorA signal worked.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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