Coding

Part:BBa_K3373004:Design

Designed by: Antonin Chevenier   Group: iGEM20_Sorbonne_U_Paris   (2020-10-16)


COP1 A. thaliana codon optimized for C. reinhardtii


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 399
    Illegal PstI site found at 576
    Illegal PstI site found at 924
    Illegal PstI site found at 939
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 399
    Illegal PstI site found at 576
    Illegal PstI site found at 924
    Illegal PstI site found at 939
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 399
    Illegal PstI site found at 576
    Illegal PstI site found at 924
    Illegal PstI site found at 939
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 399
    Illegal PstI site found at 576
    Illegal PstI site found at 924
    Illegal PstI site found at 939
    Illegal NgoMIV site found at 1229
    Illegal NgoMIV site found at 1935
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

The CDS is standardized in the Phytobrick standard for the C. reinhardtii MoClo Kit at the B3 (N-terminus coding region) position by adding BbsI sites and the corresponding fusion sites on both ends. The stop codon was removed to allow for fusion with another protein sequence and also tagging at the C-terminal position, the stop codon being provided by the part on the position B5.

Source

Pseudomonas sp. (strain ADP) 47660 [NCBI]

References

1. Crozet P, Navarro FJ, Willmund F, Mehrshahi P, Bakowski K, Lauersen KJ, Pérez-Pérez ME, Auroy P, Gorchs Rovira A, Sauret-Gueto S, Niemeyer J, Spaniol B, Theis J, Trösch R, Westrich LD, Vavitsas K, Baier T, Hübner W, de Carpentier F, Cassarini M, Danon A, Henri J, Marchand CH, de Mia M, Sarkissian K, Baulcombe DC, Peltier G, Crespo JL, Kruse O, Jensen PE, Schroda M, Smith AG, Lemaire SD. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synth Biol. 2018 Sep 21;7(9):2074-2086. doi: 10.1021/acssynbio.8b00251. Epub 2018 Sep 5. PMID: 30165733.

2. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. A modular cloning system for standardized assembly of multigene constructs. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PMID: 21364738; PMCID: PMC3041749.

3. Yamamoto YY, Matsui M, Ang LH, Deng XW. Role of a COP1 interactive protein in mediating light-regulated gene expression in arabidopsis. The Plant Cell. 1998 Jul;10(7):1083-1094. DOI: 10.1105/tpc.10.7.1083.