Composite

Part:BBa_K1602047

Designed by: Christian Sator,Stefan Zens, Max Zander, Benedikt Spannenkrebs, Sebastian Jaeger   Group: iGEM15_TU_Darmstadt   (2015-09-16)
Revision as of 19:26, 25 September 2015 by CSator (Talk | contribs)

RRK3-RRL3G

Composite part consisting of RRK3 (BBa_K1602046) cloned upstream of RRL3G (BBa_K1602045).

RRK3-RRL3G is a riboregulator-system for posttransciptional regulation of GFP-expression. The expression of GFP(BBa_E0040 is under the control of a constitutive promoter (BBa_J23100 but repressed post-transcriptional by the cis-repressing sequence (crRNA) of Lock3 (BBa_J01080). Lock3 prevents the translation of the GFP sequence by forming a hairpin-secondary-structure masking the ribosome binding site (RBS).

The presence of the trans-activating RNA-sequence (taRNA) (Key3-BBa_J01086) results in a RNA-RNA-complex and the release of the RBS enabling the translation of the GFP sequence (Fig.1).

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

The production of the taRNA is controlled by an araC-regulated pBAD-promoter (BBa_K808000). In the presence of glucose araC-pBAD shows a very low basal expression level resulting in a riboregulator-system that represses GFP expression in the presence of glucose but can be induced by the addition of arabinose.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1463
    Illegal NheI site found at 1486
  • 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 BsaI.rc site found at 2189
    Illegal SapI site found at 961


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Categories
Parameters
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