Part:BBa_K1323005:Design
The sRNA binding region.
sRNA 1 targets the YFP gene starting at the end of its Shine-Dalgarno (SD) sequence. The SD region is determined by Yoo et al. (2013) to be the most effective binding region for mRNA silencing. This 26 bp sequence has binding energy of -27.6 kcal/mol as determined using DINAMelt software (http://mfold.rna.albany.edu/?q=DINAMelt/Two-state-melting). For effective binding, sRNA should have a binding energy between -30 kcal/mol to -40 kcal/mol (Yoo et al., 2013). We aligned the sRNA 1 sequence with other constructs and the ATCC 12228 genome to determine the highest binding energy for non-specific RNA binding, which is -21.18 kcal/mol. This shows that sRNA 1 binds more specifically to YFP and should not interfere with other gene expressions in our system.
sRNA (1): YFP-targeting molecule
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 140
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]