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Composite

Part:BBa_K1602051

Designed by: Christian Sator,Stefan Zens, Max Zander, Benedikt Spannenkrebs, Sebastian Jaeger   Group: iGEM15_TU_Darmstadt   (2015-09-16)
Revision as of 06:26, 25 September 2015 by CSator (Talk | contribs)

araCpBad-RRkey

Composite part consisting of an araC-regulated pBAD-promoter (BBa_K808000) cloned upstream of RRkey (BBa_K1602049). It is half of a two-part riboregulator-system for E.coli for posttransciptional regulation of gene expression. Upon transcription the produced trans-activating RNA-sequence (taRNA) forms a RNA-RNA-complex with corresponding cis-repressing sequences (crRNA). This leads to a helix shift and the release of the formerly masked ribosome binding site (RBS) enabling the expression of the regulated gene of interest (GOI). The corresponding crRNAs are RRlocked, RRlocked_site and RRGFP.

Figure 1: Interaction of taRNA and crRNA leads to gene expression.

Because of the araC-regulated pBAD-promoter production of the taRNA is induced by the addition of arabniose. In the presence of glucose araCpBAD shows a very low basal expression level. In combination with the corresponding crRNA-parts (RRlocked and RRlocked_site) this results in an induceable riboregulator-system which represses gene expression in the presence of glucose but is inducable through the addition of arabinose.

Functional Parameters

References

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 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1215
    Illegal SapI site found at 961


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Parameters
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