Difference between revisions of "Part:BBa K1602047"

 
 
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<partinfo>BBa_K1602047 short</partinfo>
 
<partinfo>BBa_K1602047 short</partinfo>
  
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Composite part consisting of RRK3 (<html><a href="/Part:BBa_K1602046">BBa_K1602046</a></html>) cloned upstream of RRL3G (<html><a href="/Part:BBa_K1602045">BBa_K1602045</a></html>).
  
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RRK3-RRL3G is a riboregulator-system for posttransciptional regulation of GFP-expression. The expression of GFP (<html><a href="/Part:BBa_E0040">BBa_E0040</a></html>) is under the control of a constitutive promoter (<html><a href="/Part:BBa_J23100">BBa_J23100</a></html>) but repressed post-transcriptional by the cis-repressing sequence (crRNA) of Lock3 (<html><a href="/Part:BBa_J01080">BBa_J01080</a></html>). Lock3 prevents the translation of the GFP sequence by forming a hairpin-secondary-structure masking the ribosome binding site (RBS).
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The presence of the trans-activating RNA-sequence (taRNA) (<html><a href="/Part:BBa_J01086">Key3-BBa_J01086</a></html>) results in a RNA-RNA-complex and the release of the RBS enabling the translation of the GFP sequence (Fig.1).
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<figure >
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<img width=35%; src="https://static.igem.org/mediawiki/parts/3/3d/Schema_GOI.png">
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<figcaption><b>Figure 1:</b> Interaction of taRNA and crRNA leads to expression of GFP.</figcaption>
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The production of the taRNA is controlled by an araC-regulated pBAD-promoter (<html><a href="/Part:BBa_K808000">BBa_K808000</a></html>). 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.
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===Functional Parameters===
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===References===
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===Functional Parameters===
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<partinfo>BBa_K1602047 parameters</partinfo>
 
<partinfo>BBa_K1602047 parameters</partinfo>
 
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Latest revision as of 19:29, 25 September 2015

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