Coding

Part:BBa_K2273109:Design

Designed by: Nina Lautenschlaeger   Group: iGEM17_TU_Dresden   (2017-09-25)


BlaZ Beta-Lactamase derived from Staphylococcus aureus N315


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 742


Design Notes

After codon optimisation the sequence war further modified by adding a Ribosome Binding Site (AGGAGG) and a seven nucleotide spacer upstream of the start codon of the coding sequence. To enable translational fusions, e.g. with GFP (Green Fluorescent Protein), the RFC25 Restriction sites have been added 5' and 3' of the sequence:

Prefix with EcoRI, NotI, XbaI, RBS, spacer sequence, Start Codon and NgoMIV GAATTCGCGGCCGCTTCTAGAAGGAGGTGTCAAAATGGCCGGC
Suffix with AgeI, Stop Codon, SpeI, NotI and PstI ACCGGTTAAACTAGTAGCGGCCGCTGCAGA

Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI in red



Source

This part has been codon optimized for Bacillus subtilis and therefor has been chemically synthesized. The sequence is based on the S. aureus N315 genome sequence found in the NCBI database and has been codon optimized using the IDT DNA online tool.

References