Part:BBa_K2398555

Organosilicon binding Riboswitch

This in silico designed riboswitch by the MAWS software provided by the iGEM Team Heidelberg 2015 is made for the binding of Organosilicons. To be precise, ethyl 2-(dimethyl(phenyl)silyl)propanoate was used as the substrate. It can be used to 2.5-fold activate your downstream gene of interest as it is shown by our experiments. This part was ordered from IDT as oligo and cloned in pSB1C3 containing NLuc (Promega) as a reporter gene. As riboswitches are only specific for one substrate this part is only validated for the use of the appropriate substrate.

Usage and Biology

Riboswitches, in general, are used for the binding of specific molecules to release an intrinsic RBS that leads to translation of the downstream gene of interest.

Characterization

To prove that our riboswitch can activate a downstream localized gene of interest we designed a reporter system in which we cloned the in silico designed riboswitch behind a promoter region and in front of the luciferase NLuc, purchased from Promega.

Figure 1: Used educts are depicted at the top while produced products are depicted on the bottom

Figure 1: Used educts are depicted at the top while produced products are depicted on the bottom

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]