Part:BBa_K1017202

Regulation RBS-2

sRNA are small (50-250 nucleotide) non-coding RNA molecules produced by bacteria; they are highly structured and contain several stem-loops.sRNAs interact with the targeted mRNAs by imperfect base pairing, occluding the Shine-Dalgarno sequence thus prevent the ribosome from binding to the initiation condon, so the translation would be repressed.

In our project we have designed the artificial sRNA(BBa_K1017404[1]), which contains a consensus sequence, 5’-CCCUC-3’, that can base pair with the SD sequence due to complementarity, and we can get a new RBS which only would be bound with our artificial sRNA. Here we call this RBS as rRBS.

We designed the rRBS by employing the sRNA targeting region from ompF so that the sRNA would only complementary with ompF but not the others in the E.coli. We also make the AUG codon sufficiently apart from the SD sequence for ribosome binding. Therefore, other iGEM teams can use this sRNA regulation system in there project by adding this RBS to the upstream of any gene they want to regulate.

Description of function

The small RNA has three stem-loop double stranded RNA structures, and the loop which closest to the 3’ terminus is complementary to a sequence preceding the initiation codon of target mRNA. Base pairing between small RNA and target mRNA prevents the ribosome from binding to the initiation condon, so the translation would be repressed.

Acknowedgment of sources and references

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]