Difference between revisions of "Part:BBa K1075010"

 
 
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<partinfo>BBa_K1075010 short</partinfo>
 
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This part is used to induce the function of sspB with rapamycin. It was originally characterized in pSB3C5
 
This part is used to induce the function of sspB with rapamycin. It was originally characterized in pSB3C5
  
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The part was designed by Joey Davis. [1]
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The part was designed by Joey Davis.
  
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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The SspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. To control degradation it is reasonable to control the function of SspB. That is made by splitting it into two parts, each of which cannot induce degradation on its own.  To bring both SspB parts together again for inducible degradation, they were combined with a chemical inducible heterodimerisation system: FRB and FKBP12. These two parts interact in the presence of rapamycin. Linked to the parts of SspB the split system interacts when rapamycin is added and can induce degradation.
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The Plasmid pJD427 contains the fusion proteins SspB[CORE]-FRB and FKBP12-SspB[XB]:  SspB[CORE]-FRB with the weak constitutive promoter proB, FKBP12-SspB[XB] with the strong constitutive promoter proC and a medium-copy p15a origin of replication.
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1075010 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1075010 SequenceAndFeatures</partinfo>

Latest revision as of 00:53, 5 October 2013

pC-RBS32-FKBP12-EcsspB[XB]-Term-pB-RBS32-EcsspB[Core]-FRB-Term-Term (Rapamycin inducable EcsspB)

This part is used to induce the function of sspB with rapamycin. It was originally characterized in pSB3C5

The part was designed by Joey Davis.

Usage and Biology

The SspB protein is an adaptor responsible for delivering ssrA-tagged substrates to the ClpXP protease in order to enhance their degradation. To control degradation it is reasonable to control the function of SspB. That is made by splitting it into two parts, each of which cannot induce degradation on its own. To bring both SspB parts together again for inducible degradation, they were combined with a chemical inducible heterodimerisation system: FRB and FKBP12. These two parts interact in the presence of rapamycin. Linked to the parts of SspB the split system interacts when rapamycin is added and can induce degradation. The Plasmid pJD427 contains the fusion proteins SspB[CORE]-FRB and FKBP12-SspB[XB]: SspB[CORE]-FRB with the weak constitutive promoter proB, FKBP12-SspB[XB] with the strong constitutive promoter proC and a medium-copy p15a origin of replication.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1174
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 102
    Illegal BsaI.rc site found at 743