Difference between revisions of "Part:BBa K1045002"

Revision as of 08:13, 27 October 2013

CFP Reporter under control of the c-di-AMP-dependent YdaO Riboswitch

This part is a reporter system.
It is composed of a riboswitch from Bacillus subtilis genomic DNA with its native promoter and its native RBS (BBa_K1045004), controlling the expression of cfp as a reporter gene (BBa_E0020).

Ideally, in the presence of c-di-AMP, the riboswitch leads to premature termination of cfp transcription. In the absence of c-di-AMP, the system uses the terminator present in the pSB1C3 backbone resulting in formation of a cfp mRNA which can be translated in a functional CFP protein. Thus, in the abscence of c-di-AMP, the system should fluoresce. In the presence of c-di-AMP, however, the system should not fluoresce.

Furthermore, in combination with the diadenylate cyclase domain (BBa_K1045003), it could report the state of the c-di-AMP biosynthesis. This could be exploited in order to screen substances for effects on c-di-AMP homeostasis.

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]

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 It is composed of a riboswitch from ''Bacillus subtilis'' genomic DNA with its native promoter and its native RBS ([[Part:BBa_K1045004|BBa_K1045004]]), controlling the expression of ''cfp'' as a reporter gene ([[Part:BBa_E0020|BBa_E0020]]).
-Ideally, in the prescence of c-di-AMP, the riboswitch leads to premature termination of ''cfp'' transcription. In the absence of c-di-AMP, the system uses the terminator present in the '''pSB1C3''' backbone resulting in formation of a ''cfp'' mRNA which can be translated in a functional CFP protein. Thus, in the abscence of c-di-AMP, the system should fluoresce. In the presence of c-di-AMP, however, the system should not fluoresce.
+Ideally, in the presence of c-di-AMP, the riboswitch leads to premature termination of ''cfp'' transcription. In the absence of c-di-AMP, the system uses the terminator present in the '''pSB1C3''' backbone resulting in formation of a ''cfp'' mRNA which can be translated in a functional CFP protein. Thus, in the abscence of c-di-AMP, the system should fluoresce. In the presence of c-di-AMP, however, the system should not fluoresce.
 Furthermore, in combination with the diadenylate cyclase domain ([[Part:BBa_K1045003|BBa_K1045003]]), it could report the state of the c-di-AMP biosynthesis. This could be exploited in order to screen substances for effects on c-di-AMP homeostasis.