Part:BBa_K1463560

Reverse RBS

Reverse ribosome binding site designed using the Salis ribosome binding site calculator to have a strength of 12,204 when placed downstream of the reversed rfp part BBa_K1463520

Usage and Biology

This has been used to drive expression of the reversed RFP part BBa_K1463520 in our phiC31 integrase-based inversion switch BBa_K1463000.

It gave a higher level of fluorescence than a previous design for a reverse RBS BBa_I742130

Fig. 1: Red fluorescence of colonies containing BBa_K1463560 or BBa_I742130 reversed RBSs "downstream" of the reversed rfp gene BBa_K1463520 transcribed from the PhiC31 integrase switch BBa_K1463000. The image was scanned using a GE Healthcare Typhoon FLA9500 using the 532 nm laser and LPG filter.

It was designed using the Salis RBS calculator software using the RBS sequence NNNNNNNNNNCTCTAGTA (10 Ns followed by a reversed scar) upstream of the RFP open reading frame.

Fig. 2: Salis RBS calculator output for reverse RBS design

As can be seen, the designed RBS gives the start codon at position 39 a calculated translation initiation rate of "12204.43". This is 25 fold higher than the calculated translation initiation rate of BBa_I742130.

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]