Part:BBa_K2909007

ParoR_HiBiT-CloverGFP

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 1567
  Illegal PstI site found at 2846
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 1567
  Illegal NheI site found at 269
  Illegal PstI site found at 2846
  Illegal NotI site found at 1779
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 1567
  Illegal BamHI site found at 11
  Illegal BamHI site found at 3453
  Illegal XhoI site found at 2320
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 1567
  Illegal PstI site found at 2846
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 5
  Illegal EcoRI site found at 1567
  Illegal PstI site found at 2846
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 3491
  

Introduction

1- Biological background

This part is a reporter construct with a Clover GFP reporter gene tagged with a N-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Paromomycin resistance cassette called AphVIII to select the recombinant clones.

This part has been designed to be used as a reporter construct to characterize our N-terminal HiBiT tag for the Chlamydomonas reinhardtii MoClo Kit (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 restriction 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. 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).
2. 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).