Difference between revisions of "Part:BBa K2909006"

 
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
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=='''Introduction'''==
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===Functional Parameters===
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<partinfo>BBa_K2909006 parameters</partinfo>
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<h3> 1- Biological background </h3>
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This part is a reporter construct with a Clover GFP reporter gene tagged with a C-terminal NanoLuc 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.<br>
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This part has been designed to be used as a control for the characterization of our HiBiT tag parts (BBa_K2909000 and BBa_K2909001).<br>
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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.
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<h3> 2- Usage in iGEM projects </h3>
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Bio(oil)gical Factory (iGEM Sorbonne Université 2019)
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=='''Characterization'''==
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=='''References'''==
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<ol>
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<li> 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). </li>
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<li> 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).</li>
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</ol>

Latest revision as of 08:19, 29 August 2019

ParoR_CloverGFP-NanoLuc

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 2806
  • 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 2806
    Illegal NotI site found at 1779
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 1567
    Illegal BglII site found at 3629
    Illegal BamHI site found at 11
    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 2806
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5
    Illegal EcoRI site found at 1567
    Illegal PstI site found at 2806
    Illegal NgoMIV site found at 3445
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 3946


Introduction

1- Biological background

This part is a reporter construct with a Clover GFP reporter gene tagged with a C-terminal NanoLuc 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 control for the characterization of our HiBiT tag parts (BBa_K2909000 and BBa_K2909001).

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)

Characterization

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).