Reporter

Part:BBa_K3030017

Designed by: Xiaomeng Liu   Group: iGEM19_XJTLU-CHINA   (2019-09-13)
Revision as of 12:13, 14 October 2019 by LiuXiaomeng (Talk | contribs)


theophylline riboswitch with optimized RBS + eGFP

This is a part that can test the function of the optimized RBS. The theophylline riboswitch is initially designed by Markus Wieland and J�rg S. Hartig in 2008. We optimize this sequence by replacing the Shine-Dalgarno (SD) sequence with our calculated RBS by thermodynamic model. This switch is theophylline responsive which means this switch will undergo self-cleavage at the presence of theophylline. After self-cleavaging, 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 754


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