Difference between revisions of "Part:BBa K2926006"
 
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https://2019.igem.org/wiki/images/e/e8/T--Bielefeld-CeBiTec--IconAssembly.png
 
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As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA.
 
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    Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.
 
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  As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA.
 
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    Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.
 
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[[Image:T--Bielefeld-CeBiTec--IconAssembly.png|600px|thumb|center|
 
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  <b>As soon as the Cas13a:crRNA surveillance complex is formed, it can be activated by the detection of its target RNA.
 
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    Aktivation leads to colleteral cleavage of all nearby RNA strands, resulting in cell death.
 
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                          <b>Figure 1:</b> Growth curves measuring OD 600 with <i>E. coli</i> KRX BBa_K2638201 at different CuSO<sub>4</sub> concentrations. Left: No induction. Right: Induction started simultanously with inoculation with 0.1 % rhamnose and 0.1 mM IPTG.
 
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==Plasmid Design==
 
==Plasmid Design==
 
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For the analysis and characterization of the SNR52 Promotor <a href="XXX">XXXX</a>
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For the analysis and characterization of the SNR52 Promotor  
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in combiantion with the SUP4 Terminaror <a href="XXX">XXXX</a>,
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in combiantion with the SUP4 Terminaror,
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mCherry <a href="XXX">XXXX</a> was cloned beteen them using Gibson Assembly.
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mCherry was cloned beteen them using Gibson Assembly.
 
The SNR52 Promotor and SUP4 Terminator where optained as a gBlock from IDT.
 
The SNR52 Promotor and SUP4 Terminator where optained as a gBlock from IDT.
 
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We performed the analysis with <i>E. coli DH5α</i> as well as <i>S. cerevisiae</i> cultures.
 
We performed the analysis with <i>E. coli DH5α</i> as well as <i>S. cerevisiae</i> cultures.
 
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See <a href="XXX">BBa_K2926072</a> and <a href="XXX">BBa_K2926073</a> for the characterization results.
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See <a href="https://parts.igem.org/Part:BBa_K2926003">here</a> for the characterization results.
 
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Latest revision as of 03:22, 22 October 2019


SUP4 yeast terminator


Usage and Biology

SUP4 is a natural terminator of Saccharomyces cerevisiae, belonging to the SUP4 tRNATyr locus.



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]


Plasmid Design

For the analysis and characterization of the SNR52 Promotor in combiantion with the SUP4 Terminaror, mCherry was cloned beteen them using Gibson Assembly. The SNR52 Promotor and SUP4 Terminator where optained as a gBlock from IDT.


Sequencing Results

The biobrick was analysed with Sanger Sequenceing to confirm its correct base sequence.


Characterisation

We created a new protocol for the calibration of mCherry measurements with TexasRed, based on the standard iGEM protocoll for GFP fluorescence calibration, We performed the analysis with E. coli DH5α as well as S. cerevisiae cultures.
See here for the characterization results.



References