Difference between revisions of "Part:BBa K2992013"

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===Usage===
 
===Usage===
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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>pyrE</i>  operon of <i>C. sporogenes</i>  following chromosomal knock-in of <i>botR</i> ([https://parts.igem.org/Part:BBa_K2992002 BBa_K2992002]) at this locus. The secondary structure for T<i>fad</i> was predicted using the Mfold web tool (Zuker, 2003).
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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_K2992025], at this locus. The secondary structure for T<i>fad</i> was predicted using the Mfold web tool (Zuker, 2003).
 
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===Characterisation===
 
===Characterisation===
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This basic part was used in p<i>botR</i> to prevent run through expression from the pyrE operon through to the <i>botR</i> gene, therefore preventing any basal levels of BotR expression. See our [https://2019.igem.org/Team:Nottingham/Results results page] for more information.  
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This basic part was used upstream of P<i>botR</i> to prevent chromosomal read-through from the <i>pyrKDE</i> operon, upon integration of the P<i>botR</i>-<i>botR</i> module at this locus. See our [https://2019.igem.org/Team:Nottingham/Results results page] for more information.  
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<br>
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https://2019.igem.org/wiki/images/5/5e/T--Nottingham--Basic4.png
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<br>
 
<br>
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https://2019.igem.org/wiki/images/6/6b/T--Nottingham--Basic3.png
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2992013 SequenceAndFeatures</partinfo>
  
 
===References===
 
===References===

Latest revision as of 02:52, 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_K2992025, at this locus. The secondary structure for Tfad was predicted using the Mfold web tool (Zuker, 2003).

Terminator.png

Tfad from C. tetani

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

Secondary Structure

Mfold-K2992013-1.png

Characterisation

This basic part was used upstream of PbotR to prevent chromosomal read-through from the pyrKDE operon, upon integration of the PbotR-botR module at this locus. See our results page for more information.
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]

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]