Part:BBa_K1602054

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.

Figure 1: Design of RRGFP.

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. 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.

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