Difference between revisions of "Part:BBa K2384010"

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The lac repressor is a DNA-binding protein which inhibits the expression of genes coding for proteins involved in the metabolism of lactose in bacteria. These genes are repressed when lactose is not available to the cell, ensuring that the bacterium only invests energy in the production of machinery necessary for uptake and utilization of lactose when lactose is present. When lactose becomes available, it is converted into allolactose, which inhibits the lac repressor's DNA binding ability. Loss of DNA binding by the lac repressor is required for transcriptional activation of the operon.  
 
The lac repressor is a DNA-binding protein which inhibits the expression of genes coding for proteins involved in the metabolism of lactose in bacteria. These genes are repressed when lactose is not available to the cell, ensuring that the bacterium only invests energy in the production of machinery necessary for uptake and utilization of lactose when lactose is present. When lactose becomes available, it is converted into allolactose, which inhibits the lac repressor's DNA binding ability. Loss of DNA binding by the lac repressor is required for transcriptional activation of the operon.  
  
The optimization of this sequence is more suitable for the expression of <i>Bacillus megaterium</i>
 
  
 
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<table><tr>
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<th>[[Image:T--FAFU-CHINA-Toggle.png|thumb|400px|'''Figure 1''':Toggle switch. ]]</th>
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<b>Optimization Report</b>.
</tr></table>
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This part is the combination of other two parts([[Part:BBa_K2384011|BBa_K2384011]]) and ([[Part:BBa_K2384009|BBa_K2384009]]) , which could work together as a kill switch that uses a transcription-based monostable toggle design to provide rapid and robust target cell killing, dubbed ‘<b>Toggle switch</b>’
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<b>This part has made a condon optimization</b>.
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Sequence optimization information:
 
Sequence optimization information:
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Expression System: <i>Bacillus megaterium</i>
 
Expression System: <i>Bacillus megaterium</i>
  
Gene Length:624(bp)
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Gene Length: 1986 (bp)
 
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<table><tr>
 
<table><tr>
<th>[[Image:T--FAFU-CHINA-1laci.png|thumb|400px|'''Figure 4 ''':Codon Used Distribution after optimization. ]]</th>
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<th>[[Image:T--FAFU-CHINA-1mcpb.png|thumb|400px|'''Figure 4 ''':Codon Used Distribution after optimization. ]]</th>
 
</tr></table>
 
</tr></table>
  
 
<br/><br/>
 
<br/><br/>
 
<table><tr>
 
<table><tr>
<th>[[Image:T--FAFU-CHINA-2laci.png|thumb|400px|'''Figure  5''':GC Content after optimization. ]]</th>
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<th>[[Image:T--FAFU-CHINA-2mcpb.png|thumb|400px|'''Figure  5''':GC Content after optimization. ]]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
<th>[[Image:T--FAFU-CHINA-dnalaci.png|thumb|400px|'''Figure 3''':  Digested with NotI//. ]]</th>
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<th>[[Image:T--FAFU-CHINA-dnamcpb.png|thumb|400px|'''Figure 3''':  Digested with NotI/KpnI. ]]</th>
 
</tr></table>
 
</tr></table>
  

Revision as of 17:19, 1 November 2017


LacI

This part is combined with PLtetO-1 promoter+RBS+CDS+lambda t0 terminator.

Usage and Biology

The lac repressor is a DNA-binding protein which inhibits the expression of genes coding for proteins involved in the metabolism of lactose in bacteria. These genes are repressed when lactose is not available to the cell, ensuring that the bacterium only invests energy in the production of machinery necessary for uptake and utilization of lactose when lactose is present. When lactose becomes available, it is converted into allolactose, which inhibits the lac repressor's DNA binding ability. Loss of DNA binding by the lac repressor is required for transcriptional activation of the operon.




Optimization Report.

Sequence optimization information:

Expression System: Bacillus megaterium

Gene Length: 1986 (bp)

Show the relative codon used distribution


Figure 4 :Codon Used Distribution after optimization.



Figure 5:GC Content after optimization.
Figure 3: Digested with NotI/KpnI.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 115
  • 1000
    COMPATIBLE WITH RFC[1000]