Part:BBa_K2800031

RBS strength is 10

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Biology:

    Ribosomal binding sites refer to a purine rich untranslated region upstream of the start codon AUG. Also known as SD (Shine-Dalg-arno) sequence. In RBS, there is a sequence of SD (Shine-Dalg-arno) with a length of 5 nucleotides, which is rich in G and A. The sequence complements with the 3'end of ribosome 16S rRNA, facilitating the binding of ribosomes to the mRNA and facilitating the initiation of translation. The binding strength of RBS depends on the structure of SD sequence and the distance between SD sequence and AUG.

Characterization:

    This RBS is predicted to be very suitable to Zymomonas mobilis by the algorithm of SALIS with Zymomonas mobilis’s 16sRNA. We test this RBS with our dual reporter-gene system in Zymomonas mobilis, and the relativity of experimental results is relatively high with predicted results.(Fig. 1) And we also test these RBS in E.coli and the results shows that they are also suitable to E.coli. And it will inhibit the growth of E.coli when Tc concentration is more than 0.2 ug/mL, so the ratio of EGFP/mCherry gets down when Tc concentration increased.(Fig.2) And we also measured the correlation of Tc concentration with RBS strength in Z. mobilis.

Fig.1 Different RBS strength in Zymomonas mobilis.

Fig.2 Different RBS strength in E.coli.

Fig.3 Correlation of Tc concentration with RBS strength in Z. mobilis

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 7
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 7
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 7
  
• 1000
  COMPATIBLE WITH RFC[1000]