Difference between revisions of "Part:BBa K1913007"

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This part contains two viral genes: 434- and lambda cI (BBa_C0052 and part of BBa_K081007) expressed under the pBAD/AraC promoter (BBa_I0500). The 434 cI gene is not yet preceded by an RBS, allowing tuning of the cI protein balance. The 434 cI repressor protein encoded in this part should be more rapidly degraded (see design considerations) than the lambda cI protein.  
 
This part contains two viral genes: 434- and lambda cI (BBa_C0052 and part of BBa_K081007) expressed under the pBAD/AraC promoter (BBa_I0500). The 434 cI gene is not yet preceded by an RBS, allowing tuning of the cI protein balance. The 434 cI repressor protein encoded in this part should be more rapidly degraded (see design considerations) than the lambda cI protein.  
 
Therefore, changing the strength of the pBAD promoter in this part (with L-Arabinose or D-Glucose) should temporary shift the ratio between the two cI protein levels. We recommend using this part together with the modified lambda Prm promoter (BBa_I12006). Combining these parts should lead to interesting dynamics of expression under this modified lambda Prm promoter in response to changes in pBAD promoter strength.
 
Therefore, changing the strength of the pBAD promoter in this part (with L-Arabinose or D-Glucose) should temporary shift the ratio between the two cI protein levels. We recommend using this part together with the modified lambda Prm promoter (BBa_I12006). Combining these parts should lead to interesting dynamics of expression under this modified lambda Prm promoter in response to changes in pBAD promoter strength.
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I suspect the viral genes slightly impair growth of DH5alpha. Growth was found to be disappointing multiple times, but this has not been quantified.
  
  
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The 434 cI protein encoded in this part contains a C-terminal LVA tag causing more rapid protein degradation, the lambda cI gene in this part does not encode an LVA tag (although BBa_K081007 does contain an LVA tag).
 
  
 
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>===Sequence and Features===</span>
 
<partinfo>BBa_K1913007 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1913007 SequenceAndFeatures</partinfo>
  

Revision as of 12:15, 8 October 2016


434- and lambda cI operon for tuning protein balance

This part is intended to provide a balance between the 434- and lambda cI proteins depending on the strength of the RBS that is inserted upstream of 434 cI.


Usage and Biology

This part contains two viral genes: 434- and lambda cI (BBa_C0052 and part of BBa_K081007) expressed under the pBAD/AraC promoter (BBa_I0500). The 434 cI gene is not yet preceded by an RBS, allowing tuning of the cI protein balance. The 434 cI repressor protein encoded in this part should be more rapidly degraded (see design considerations) than the lambda cI protein. Therefore, changing the strength of the pBAD promoter in this part (with L-Arabinose or D-Glucose) should temporary shift the ratio between the two cI protein levels. We recommend using this part together with the modified lambda Prm promoter (BBa_I12006). Combining these parts should lead to interesting dynamics of expression under this modified lambda Prm promoter in response to changes in pBAD promoter strength. I suspect the viral genes slightly impair growth of DH5alpha. Growth was found to be disappointing multiple times, but this has not been quantified.




===Sequence and Features===


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1898
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961