Composite

Part:BBa_K4191003:Design

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


J23119 + B0034 + HAD + B0010


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 57
    Illegal BsaI.rc site found at 831


Design Notes

While designing the sequence in SnapGene, we had to remove certain restriction sites (BsaI, Sapl) from our gene sequence (HAD), 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. The other iGEM designed parts had their overhangs modified to fit the respective A-B (promoter), B-C (RBS), and D-F (terminator) overhangs for the u loop system.

Source

This part is composed of a promoter (BBa_J23119), RBS (BBa_B0034), gene sequence (BBa_K4191004), and terminator (BBa_B0010). The gene sequence of haloacid dehalogenase (HAD) is derived from Xanthobacter autotrophicus and has been codon optimized for E. Coli.


References