Difference between revisions of "Part:BBa K2992013"

Revision as of 01:30, 22 October 2019

Usage

Tfad is a terminator sequence derived from the fad gene of C. tetani, encoding FAD-binding oxidreducatse. These enzymes catalyse the conversion of dihydroxyacetone phosphate to glycerol-3-phosphate in a reversible reaction. In our project we use this strong terminator to prevent polar transcription from the pyrKDE operon of C. sporogenes, following chromosomal knock-in of botR in part BBa_K29920252, at this locus. The secondary structure for Tfad was predicted using the Mfold web tool (Zuker, 2003).

Tfad from C. tetani

Transcriptional terminator sequence from the FAD-dependent oxidoreductase, fad, from Clostridium tetani

Secondary Structure

Characterisation

This basic part was used in pbotR to prevent run through expression from the pyrE operon through to the botR gene, therefore preventing any basal levels of BotR expression. See our results page for more information.

References

Zuker, M. (2003). Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Research, 31(13), pp.3406-3415.

Measurement

[http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]

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 ===Usage===
-T<i>fad</i>  is a terminator sequence derived from the <i>fad</i> gene of <i>C. tetani</i>, encoding FAD-binding oxidreducatse. These enzymes catalyse the conversion of dihydroxyacetone phosphate to glycerol-3-phosphate in a reversible reaction. In our project we use this strong terminator to prevent polar transcription from the <i>pyrKDE</i>  operon of <i>C. sporogenes</i>,  following chromosomal knock-in of <i>botR</i> in part [https://parts.igem.org/Part:BBa_K2992025 BBa_K29920252]), at this locus. The secondary structure for T<i>fad</i> was predicted using the Mfold web tool (Zuker, 2003).
+T<i>fad</i>  is a terminator sequence derived from the <i>fad</i> gene of <i>C. tetani</i>, encoding FAD-binding oxidreducatse. These enzymes catalyse the conversion of dihydroxyacetone phosphate to glycerol-3-phosphate in a reversible reaction. In our project we use this strong terminator to prevent polar transcription from the <i>pyrKDE</i>  operon of <i>C. sporogenes</i>,  following chromosomal knock-in of <i>botR</i> in part [https://parts.igem.org/Part:BBa_K2992025 BBa_K29920252], at this locus. The secondary structure for T<i>fad</i> was predicted using the Mfold web tool (Zuker, 2003).
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