Part:BBa_K2276008

a composite of TetR repressible promoter、special designed RBS and the coding sequence of mRFP

The part was designed based on the Part:BBa_I13521,while the RBS binding site is modified. The RBS is special designed for strains whose last 9 nucleotides of the 16S rRNA is ACCTCCTTA.We want to generate a part which present a high translation rate, but this one did not meet our expectation.The promoter is TetR repressible promoter which is also used in the repressilator system. And the mRFP served as the reporter to help us measure the translation rate of the designed RBS.

Design Notes

We want to engineer a strain which can produce melatonin at daily rhythm.We employ repressilator system thus allowing the E.coli coding enzyme periodically. However, the cycle time is not 24h.We built a model to analyse the issue.According to the medeling result, a stronger RBS was needed.We turned to the bioinformatic methods to design a RBS predicted to show hign translation rate.This is the website on which we performed the design: https://salislab.net/software/doForwardRBS. We designed three, and BBa_K2276007 is one of them.

Fig 1.The design result of the RBS in Part:BBa K2276007.

Fig 2.The prediction of the translation rate of Part:BBa I13521

Source

Part:BBa_I13521

References

1.Borujeni A E, Salis H M. Translation Initiation is Controlled by RNA Folding Kinetics via a Ribosome Drafting Mechanism[J]. Journal of the American Chemical Society, 2016, 138(22):7016.

2.https://parts.igem.org/wiki/index.php?title=Part:BBa_I13521

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 634
  Illegal AgeI site found at 746
  
• 1000
  COMPATIBLE WITH RFC[1000]