Composite

Part:BBa_K2909014:Design

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

DGAT-1-2-HiBiT_LPAAT-A-HiBiT_HygroR E. guineensis


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 3079
    Illegal EcoRI site found at 5568
    Illegal PstI site found at 2349
    Illegal PstI site found at 4307
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 3079
    Illegal EcoRI site found at 5568
    Illegal NheI site found at 5832
    Illegal PstI site found at 2349
    Illegal PstI site found at 4307
    Illegal NotI site found at 217
    Illegal NotI site found at 3291
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 3079
    Illegal EcoRI site found at 5568
    Illegal BglII site found at 2257
    Illegal BamHI site found at 4149
    Illegal XhoI site found at 758
    Illegal XhoI site found at 980
    Illegal XhoI site found at 3832
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 3079
    Illegal EcoRI site found at 5568
    Illegal PstI site found at 2349
    Illegal PstI site found at 4307
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 3079
    Illegal EcoRI site found at 5568
    Illegal PstI site found at 2349
    Illegal PstI site found at 4307
    Illegal NgoMIV site found at 4862
    Illegal NgoMIV site found at 6533
    Illegal NgoMIV site found at 6715
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2550
    Illegal SapI.rc site found at 5039


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 DGAT-1-2 enzyme (BBa_K2909003) comes from the predicted sequence from the E. guineensis genome sequencing.
NCBI Reference Sequence: XM_010933834.1
https://www.ncbi.nlm.nih.gov/nuccore/XM_010933834.1

The C-terminal HiBiT tag (BBa_K2909001) 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).