Difference between revisions of "Part:BBa K2909009"
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__NOTOC__
 
__NOTOC__
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<partinfo>BBa_K290900 short</partinfo>
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<partinfo>BBa_K2909009 short</partinfo>
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<h3> 1- Biological background </h3>
 
<h3> 1- Biological background </h3>
  
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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.<br>
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This part is a CDS coding for the E. guineensis LPAAT-A enzyme tagged with a N-terminal HiBiT tag under the control of an ammonium-controlled Chlamydomonas reinhardtii promoter called NIT1. It also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.<br>
  
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This part has been designed to characterize the expression of the E. guineensis LPAAT-A enzyme that we adapted for C. reinhardtii with the N-terminal HibiT tag(BBa_K2909002).<br>
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This part has been designed to characterize the expression of the E. guineensis LPAAT-A enzyme that we adapted for C. reinhardtii (BBa_K2909002) using the N-terminal HiBiT tag (BBa_K2909000).<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 restriciton enzyme. It is this linear DNA that is integrated into the genomic DNA of Chlamydomonas reinhardtii during the transformation.
+
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.
  
 
<h3> 2- Usage in iGEM projects </h3>
 
<h3> 2- Usage in iGEM projects </h3>
  
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Bio[oil]gical Factory (iGEM Sorbonne Université 2019)
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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> Schwinn, M. K. et al. CRISPR-Mediated Tagging of Endogenous Proteins with a Luminescent Peptide. ACS Chem. Biol. 13, 467–474 (2018). </li>
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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 08:35, 29 August 2019

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 an ammonium-controlled 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) using 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 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. 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).