Regulatory

Part:BBa_K4825020

Designed by: Qiu Tianhou   Group: iGEM23_GreatBay-SCIE   (2023-10-12)
Revision as of 12:26, 12 October 2023 by Registry (Talk | contribs)

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pAra3

pAra3 is a synthetic inducible promoter designed for automation for yeast, in which the level of the inducer arabinose regulates the initiation of the transcription. AraC functions as a homodimer. In the absence of arabinose, the DNA-binding domain of two subunits binds to Inv AraI1 and AraO2 respectively. Inv represents the inversion of origin element sequence. The two subunits are connected through the dimerisation domain. The 193base pair(bp) intervals and dimerisation enables the DNA loop to form. DNA loop hindered the binding of RNA polymerase. One more copy of AraI and AraO2 are added to this part, thus double DNA loop are expected to be formed to enhance the repression. In the presence of arabinose, two subunits of AraC primarily bind to adjacent Inv Aral1 and Inv Ara2 separated by four bp instead of looping. One more AraC is involved with the same mechanism due to the dublication. The promoter thus becomes accessible, increasing the downstream gene transcription level. In our project, this part is utilized to construct a plasmid with Bl-1 gene and control its transcription by inducing it with the presence of arabinose in S.cerevisiae and prevent the initiation of kill switch.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 39
    Illegal BamHI site found at 102
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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