Part:BBa_K2909009

HiBiT-LPAAT-A_HygroR E. guineensis

Sequence and Features

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
  

Introduction

1- Biological background

This part is a CDS coding for the E. guineensis LPAAT-A enzyme tagged with a N-terminal HiBiT tag under the control of a nitrate-inducible Chlamydomonas reinhardtii promoter called NIT1. It also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.

This part has been designed to characterize the expression of the E. guineensis LPAAT-A enzyme that we adapted for C. reinhardtii (BBa_K2909002) with the N-terminal HibiT tag (BBa_K2909000).

This part is a linear DNA that is the result of the digestion of the level M plasmid by the type IIS BsaI restriciton enzyme. It is this linear DNA that is integrated into the genomic DNA of Chlamydomonas reinhardtii during the transformation.

2- Usage in iGEM projects

Bio[oil]gical Factory (iGEM Sorbonne Université 2019)

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).