Part:BBa_K3425033

iGEM Type IIS pSB1K03-DY

This part (BBa_K3425025) is an RBS (<a href="https://parts.igem.org/wiki/index.php/Part:BBa_B0030">BBa_B0030</a>) modified for Type IIS iGEM Standard assembly (RFC1000) by adding a five-nucleotide spacer (TAGTA) to the end of the sequence.

Whereas BioBrick assembly generates six-nucleotide scar between RBS and ATG, Type IIS iGEM Standard assembly generates only one-nucleotide scar. Five nucleotides added to BBa_B0030 together with one-nucleotide scar of Type IIS assembly form the same distance between RBS and ATG as in BioBrick assembly.

The strength of RBS depends on the 1) sequence, but also 2) aligned spacing and 3) genetic context.

Aligned spacing is the distance between the centre of Shine-Dalgarno sequence and ATG start codon (see https://parts.igem.org/Ribosome_Binding_Sites/Design or Ma et al. (2002)). Aligned spacing generally varies from 5 to 13 bases, with optimum for E. coli

Five to six nucleotides is the optimal distance between the RBS and the ATG start codon for E. coli (see https://parts.igem.org/Ribosome_Binding_Sites/Design for more general information). Whereas Biobrick assembly generates six-nucleotide scar, Type IIS iGEM Standard assembly generates only one-nucleotide scar between RBS and ATG. To keep the distance optimal, five nucleotides were added to the BBa_B0030 for its use in Type IIS assembly.

Strength of an RBS is dependent on the sequence similarity of

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]