Difference between revisions of "Part:BBa K1602054"
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<partinfo>BBa_K1602054 short</partinfo>
 
<partinfo>BBa_K1602054 short</partinfo>
  
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This part is half of a two-part riboregulator-system for posttransciptional regulation of GFP-expression. The part was created by inserting a GFP-gene flanked by the restriction sites BamHI and HindIII into <html><a href="/Part:BBa_K1602053">RRlocked_site</a></html> via restriction cloning. It consists of the fused sequences of a constitutive promoter (<html><a href="/Part:BBa_J23100">BBa_J23100</a></html>), a cis-repressing sequence (cr), GFP (<html><a href="/Part:BBa_E0040">BBa_E0040</a></html>) and a terminator (<html><a href="/Part:BBa_B0015">BBa_B0015</a></html>). The restriction sites upstream and downstream of the GFP sequence allow for an easy exchange of the GFP gene with another gene of interest.
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Upon transcription the cis-repressing sequence forms a hairpin-secondary-structure masking the ribosome binding site (RBS) and therefore prevents translation of the following GFP-sequence.
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<figure >
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<img width=40%; src="https://static.igem.org/mediawiki/parts/e/e6/RRGFP.png">
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<figcaption><b>Figure 1:</b> Design of RRGFP.</figcaption>
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If the corresponding trans-activating RNA-sequence (taRNA) (<html><a href="/Part:BBa_K1602049">RRKey-BBa_K1602049</a></html>) is present the two sequences form a RNA-RNA-complex. This leads to a helix shift and the release of the RBS enabling the expression of GFP as the gene of interest (GOI).
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<center>
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<figure >
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<img width=30%; src="https://static.igem.org/mediawiki/parts/3/3d/Schema_GOI.png">
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<figcaption><b>Figure 2:</b> Interaction of taRNA and crRNA leads to expression of GFP.</figcaption>
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===Functional Parameters===
 
===Functional Parameters===

Revision as of 06:12, 25 September 2015

RRGFP

This part is half of a two-part riboregulator-system for posttransciptional regulation of GFP-expression. The part was created by inserting a GFP-gene flanked by the restriction sites BamHI and HindIII into RRlocked_site via restriction cloning. It consists of the fused sequences of a constitutive promoter (BBa_J23100), a cis-repressing sequence (cr), GFP (BBa_E0040) and a terminator (BBa_B0015). The restriction sites upstream and downstream of the GFP sequence allow for an easy exchange of the GFP gene with another gene of interest. Upon transcription the cis-repressing sequence forms a hairpin-secondary-structure masking the ribosome binding site (RBS) and therefore prevents translation of the following GFP-sequence.

Figure 1: Design of RRGFP.

If the corresponding trans-activating RNA-sequence (taRNA) (RRKey-BBa_K1602049) is present the two sequences form a RNA-RNA-complex. This leads to a helix shift and the release of the RBS enabling the expression of GFP as the gene of interest (GOI).

Figure 2: Interaction of taRNA and crRNA leads to expression of GFP.

Functional Parameters

References

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 91
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 737