Coding

Part:BBa_K4770022:Design

Designed by: Andrea Camí Bonet   Group: iGEM23_Barcelona-UB   (2023-10-12)
Revision as of 09:24, 12 October 2023 by Andreacami (Talk | contribs)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 958
    Illegal PstI site found at 2090
    Illegal PstI site found at 2511
    Illegal PstI site found at 2535
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 958
    Illegal PstI site found at 2090
    Illegal PstI site found at 2511
    Illegal PstI site found at 2535
    Illegal NotI site found at 838
    Illegal NotI site found at 2471
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3202
    Illegal BamHI site found at 1830
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 958
    Illegal PstI site found at 2090
    Illegal PstI site found at 2511
    Illegal PstI site found at 2535
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 958
    Illegal PstI site found at 2090
    Illegal PstI site found at 2511
    Illegal PstI site found at 2535
    Illegal NgoMIV site found at 888
    Illegal NgoMIV site found at 1902
    Illegal NgoMIV site found at 2168
    Illegal NgoMIV site found at 2572
    Illegal NgoMIV site found at 2584
    Illegal NgoMIV site found at 3018
  • 1000
    COMPATIBLE WITH RFC[1000]

Source of this part

Original PPSAD sequence: BBa_K4770007 Original IPT sequence: BBa_K4770005 Original F2A sequence: BBa_K4770012 Original NanoLuc sequence: BBa_K4770011 Original TPSAD sequence: BBa_K4770008

Design considerations

We performed Chlamydomonas reinhardtii's domestication for GS sequence. This process includes a reduction of the GC% content to make the sequence suitable for commercial synthesis while maintaining it high enough to fit Chlamydomonas reinhardtii's codon usage. This was done using our optimizing software (see AlgaGenix's wiki for additional information). Moreover, recognition sites for BbsI and BsaI were eliminated, changing said codons with synonymous ones. Besides, we also added an intron to perform what is known as intron-mediated enhancement (IME), which studies show aids with stable high-level expression of a foreign gene (Lumbreras et al., 1998) AlgaGenix’s Level 1s are designed to have the same structure and not having to build different pieces with different positions for each gene.