Coding

Part:BBa_K5222007:Design

Designed by: Maria Avtodeeva   Group: iGEM24_Sorbonne-U-Paris   (2024-10-02)


tphAa,tphAb,tphAc,tphB fusion protein adapted for chlamydomonas reinhardtii


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 93
    Illegal PstI site found at 246
    Illegal PstI site found at 2073
    Illegal PstI site found at 2106
    Illegal PstI site found at 2886
    Illegal PstI site found at 3063
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1104
    Illegal PstI site found at 93
    Illegal PstI site found at 246
    Illegal PstI site found at 2073
    Illegal PstI site found at 2106
    Illegal PstI site found at 2886
    Illegal PstI site found at 3063
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1432
    Illegal BglII site found at 1723
    Illegal XhoI site found at 1482
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 93
    Illegal PstI site found at 246
    Illegal PstI site found at 2073
    Illegal PstI site found at 2106
    Illegal PstI site found at 2886
    Illegal PstI site found at 3063
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 93
    Illegal PstI site found at 246
    Illegal PstI site found at 2073
    Illegal PstI site found at 2106
    Illegal PstI site found at 2886
    Illegal PstI site found at 3063
    Illegal NgoMIV site found at 561
    Illegal NgoMIV site found at 789
    Illegal NgoMIV site found at 999
    Illegal NgoMIV site found at 2448
    Illegal NgoMIV site found at 3186
    Illegal NgoMIV site found at 3641
    Illegal AgeI site found at 1450
    Illegal AgeI site found at 2716
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

To introduce this CDS into Chlamydomonas, we will employ the MoClo technique to construct the necessary plasmid for transformation. Our CDS needs to be adapted for MoClo assembly, so we plan to insert our gene into positions B3-B5 of the MoClo system. To do this, we added appropriate fusion sites at the ends of the sequence. At the 5' end, we incorporated a B3 scar (AATG) along with the BbsI restriction site, and at the 3' end, a B5 scar with a BbsI restriction site (see Chlamydomonas Guide and Pierre Crozet et al. for more details).


Source

Characterization of the Terephthalate Degradation Genes of Comamonas sp. Strain E6, [1] [1] Sasoh M, Masai E, Ishibashi S, Hara H, Kamimura N, Miyauchi K, Fukuda M. Characterization of the terephthalate degradation genes of Comamonas sp. strain E6. Appl Environ Microbiol. 2006 Mar;72(3):1825-32. doi: 10.1128/AEM.72.3.1825-1832.2006. PMID: 16517628; PMCID: PMC1393238.

References