Composite
ExpSubE

Part:BBa_K2020002:Design

Designed by: Nicola Freyer, Lea Steinbeck, Svenja Meyer, Alexander Deitert   Group: iGEM16_Aachen   (2016-10-05)


expression system for subtilisin E in E. coli


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 NgoMIV site found at 280
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The sequence of the subtilisin E gene from Bacillus subtilis was codon optimized for E. coli with the DNA and protein sequence analysis software "Geneious" and additionally with the "Codon Optimization Tool" from IDT. The sequence was partly ordered from IDT (BBa_K2020001 + BBa_B0010) and then cloned into BBa_J04500, a protein expression backbone which already carries the LacI promoter BBa_R0010 and the ribosome binding site BBa_B0034.


Source

The sequences of the BioBrick parts were obtained through the Registry.

part BioBrick No.
LacI promoter BBa_R0010
RBS BBa_B0034
CDS BBa_K2020001
BBa_J32015 BBa_K2020000
terminator BBa_B0010

The construct was ordered at IDT.