Part:BBa_K2909009:Design

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
  

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).