Coding

Part:BBa_K4191001:Design

Designed by: Ananya Bharathwaj, Madison Yang   Group: iGEM22_WVHS_SanDiego   (2022-10-09)


FAcD CDS derived from R. palustris, codon optimized for E. coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 533
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 723
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 10
    Illegal BsaI.rc site found at 931


Design Notes

While designing the sequence in SnapGene, we had to remove certain restriction sites (BsaI, Sapl) from our gene sequences, as the enzymes would digest and reassemble the sequence. Our enzyme sequence was also codon optimized for E. coli, due to its similarity to our model organism P. putida. 20 bases of homology were also added to the end of the gene sequence prior to ordering the sequence from IDT, to ensure maximum efficiency. In order to ensure the sequence was compatible with our u loop system, the overhangs were modified to match with the C-D overhangs specifically for loop assembly.


Source

Rhodopseudomonas palustris FAcD CDS

References