Difference between revisions of "Part:BBa J58100:Design"
 
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===Design Notes===
 
===Design Notes===
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We take the lambda Prm promoter sequence from the Registry, and we add the CRP binding site upstream of that promoter. The centered position of the cI operator is -42, and for the CRP is -93,5.
+
We take the lambda Prm promoter sequence from the Registry (<partinfo>I12007</partinfo>), and we added the CRP binding site upstream of that promoter. Following Joung et al. the centered position of the cI operator was fixed to -42, and for the CRP to -93.5. This forced us to do a lot of cut-and-paste of the original Prm sequence to create (presumably) harmless spacers between the operator regions. Unfortunately the CRP consensus binding site contained a XbaI site. Further work is needed to do mutagenesis on this site while preserving the binding affinity.
  
 
===Source===
 
===Source===
  
−
The sources of this promoter are, on the one hand, from the bacteriophage lambda, and, on the other hand, from the E. coli. We take the lambda Prm promoter (modified to be activated but not repressed    by cI, part ref. BBa_I12007), and the binding site region for CRP from E. coli.  
+
The sources of this promoter are, on the one hand, from the bacteriophage lambda, and, on the other hand, from the E. coli. We take the lambda Prm promoter (modified to be activated but not repressed    by cI, part ref. <partinfo>BBa_I12007</partinfo>), and the binding site region for CRP from E. coli.  
  
 
===References===
 
===References===
  
 
JK Joung, DM Koepp, A Hochschild. Synergistic activation of transcription by bacteriophage lambda cI protein and E. coli cAMP receptor protein. Science (1994), 265, 1863-1866.
 
JK Joung, DM Koepp, A Hochschild. Synergistic activation of transcription by bacteriophage lambda cI protein and E. coli cAMP receptor protein. Science (1994), 265, 1863-1866.
 +
 +
Bintu et al., Transcriptional regulation by the numbers: applications. Current Opinion in Genetics & Development 2005, 15:125–135

Latest revision as of 04:29, 30 October 2006

AND-type promoter synergistically activated by cI and CRP


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 11
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 11
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 11
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We take the lambda Prm promoter sequence from the Registry (BBa_I12007), and we added the CRP binding site upstream of that promoter. Following Joung et al. the centered position of the cI operator was fixed to -42, and for the CRP to -93.5. This forced us to do a lot of cut-and-paste of the original Prm sequence to create (presumably) harmless spacers between the operator regions. Unfortunately the CRP consensus binding site contained a XbaI site. Further work is needed to do mutagenesis on this site while preserving the binding affinity.

Source

The sources of this promoter are, on the one hand, from the bacteriophage lambda, and, on the other hand, from the E. coli. We take the lambda Prm promoter (modified to be activated but not repressed by cI, part ref. BBa_I12007), and the binding site region for CRP from E. coli.

References

JK Joung, DM Koepp, A Hochschild. Synergistic activation of transcription by bacteriophage lambda cI protein and E. coli cAMP receptor protein. Science (1994), 265, 1863-1866.

Bintu et al., Transcriptional regulation by the numbers: applications. Current Opinion in Genetics & Development 2005, 15:125–135