Part:BBa_K3030016

theophylline riboswitch with SD RBS + eGFP

This is a part that can test the function of the SD RBS. The theophylline riboswitch is initially designed by Wieland, M. and Hartig, J. S. (2008). This switch is theophylline responsive which means this switch will undergo self-cleavage at the presence of theophylline. After self-cleavaging, Shine–Dalgarno (SD) sequence will be exposed and ribosomes can bind with mRNA and translation will initiate. We use eGFP(BBa_E0040) as our reporter. The fluorescence intensity/OD. 600 can indicate the functionality of this switch. We place this part at the downstream of pBAD promoter. With the induce of L-arabinose, BL21(DE3) cells will transcribe mRNAs containing theophylline switch and eGFP coding sequence. At the presence of theophylline, eGFP protein will be translated and by detecting the fluorescence intensity/OD. 600, the functionality of this switch can be characterized.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 753
  

Reference

Wieland, M., Hartig, J.S. (2008). ‘Improved aptazyme design and in vivo screening enable riboswitching in bacteria.’ Angewandte Chemie, 47(14), pp2604-2607. [Online]. Available from: https://www.ncbi.nlm.nih.gov/pubmed/18270990