Difference between revisions of "Part:BBa K1893015"
 
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<partinfo>BBa_K1893015 short</partinfo>
 
<partinfo>BBa_K1893015 short</partinfo>
  
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A construct for controlling the expression of a gene in front of the pBAD promoter.
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This composite part is composed of a pBAD arabinose inducible promoter system developed by the 2014 Imperial team [https://parts.igem.org/Part:BBa_K1321333 (BBa_K1321333)] and an upstream reverse terminator [https://parts.igem.org/Part:BBa_B0025 (BBa_B0025)] to terminate transcription of an antisense encoded araC transcription factor in the pBAD system. This part enables arabinose inducible expression of a downstream coding sequence, and can be switched off by the addition of D-glucose.
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<!-- Add more about the biology of this part here
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===Usage and Biology===
 
===Usage and Biology===
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Without any arabinose in the system, araC forms a homodimer, binding to operator sites upstream of the pBAD promoter, generating an inhibitory loop in the DNA that prevents access by RNAP. In the absence of glucose, and the presence of L-arabinose, the dimer dissociates from the DNA, allowing transcription of whatever is downstream of the pBAD promoter. We have characterised the activation range of this part by using GFP as a reporter construct. You can find the information here.
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When arabinose is absent, araC forms a homodimer and binds to operator sites upstream of the pBAD promoter. This generates an inhibitory DNA loop that blocks RNA polymerase binding and prevents transcription. In the presence of L-arabinose, the dimer dissociates from the DNA, allowing transcription of the coding sequence downstream of the pBAD promoter. The promoter can then be switched off via catabolite repression by adding D-glucose to the system.
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We have characterised the activation range of this part using GFP as a reporter. The results of our these characterisation experiments can be found here [https://parts.igem.org/Part:BBa_K1893017 (BBa_K1893017)].
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K1893015 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1893015 SequenceAndFeatures</partinfo>

Latest revision as of 20:00, 28 October 2016


Arabinose inducible promoter (pBAD)

This composite part is composed of a pBAD arabinose inducible promoter system developed by the 2014 Imperial team (BBa_K1321333) and an upstream reverse terminator (BBa_B0025) to terminate transcription of an antisense encoded araC transcription factor in the pBAD system. This part enables arabinose inducible expression of a downstream coding sequence, and can be switched off by the addition of D-glucose.


Usage and Biology

When arabinose is absent, araC forms a homodimer and binds to operator sites upstream of the pBAD promoter. This generates an inhibitory DNA loop that blocks RNA polymerase binding and prevents transcription. In the presence of L-arabinose, the dimer dissociates from the DNA, allowing transcription of the coding sequence downstream of the pBAD promoter. The promoter can then be switched off via catabolite repression by adding D-glucose to the system.


We have characterised the activation range of this part using GFP as a reporter. The results of our these characterisation experiments can be found here (BBa_K1893017).


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1342
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1281
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1116
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1098