Part:BBa_K2909012:Design

HiBiT-DGAT-1-2_HygroR E. guineensis

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 3104
  Illegal PstI site found at 2389
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 3104
  Illegal NheI site found at 3368
  Illegal PstI site found at 2389
  Illegal NotI site found at 217
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 3104
  Illegal BglII site found at 2297
  Illegal BamHI site found at 2537
  Illegal XhoI site found at 758
  Illegal XhoI site found at 1020
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 3104
  Illegal PstI site found at 2389
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 3104
  Illegal PstI site found at 2389
  Illegal NgoMIV site found at 4069
  Illegal NgoMIV site found at 4251
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 2575
  

Source

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).