Difference between revisions of "Part:BBa K1602047"

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

@@ Line 4: / Line 4: @@
 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>).
-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).
+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).
 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).