Composite

Part:BBa_K2909009:Design

Designed by: Laurent CHEN   Group: iGEM19_Sorbonne_U_Paris   (2019-08-28)


HiBiT-LPAAT-A_HygroR E. guineensis


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 2519
    Illegal PstI site found at 1273
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 2519
    Illegal NheI site found at 2783
    Illegal PstI site found at 1273
    Illegal NotI site found at 217
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 2519
    Illegal BamHI site found at 1115
    Illegal BamHI site found at 1952
    Illegal XhoI site found at 758
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 2519
    Illegal PstI site found at 1273
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 2519
    Illegal PstI site found at 1273
    Illegal NgoMIV site found at 1828
    Illegal NgoMIV site found at 3484
    Illegal NgoMIV site found at 3666
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1990


Design Notes

Source

The LPAAT-A enzyme (BBa_K2909002) comes from the predicted sequence from the E. guineensis genome sequencing.
NCBI Reference Sequence: XM_010908457.2
https://www.ncbi.nlm.nih.gov/nuccore/1130625520

The N-terminal HiBiT tag (BBa_K2909000) comes from Promega (Schwinn et al. 2018).

All the other parts comes from the Chlamydomonas reinhardtii MoClo Kit (Crozet et al. 2018).

References

  1. Schwinn, M. K. et al. CRISPR-Mediated Tagging of Endogenous Proteins with a Luminescent Peptide. ACS Chem. Biol. 13, 467–474 (2018).
  2. Crozet, P. et al. Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synth. Biol. 7, 2074–2086 (2018).
  3. 1.Weber, E., Engler, C., Gruetzner, R., Werner, S. & Marillonnet, S. A Modular Cloning System for Standardized Assembly of Multigene Constructs. PLoS ONE 6, e16765 (2011).