Difference between revisions of "Part:BBa K2909013"
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<h3> 1- Biological background </h3> | <h3> 1- Biological background </h3> | ||
| − | This part is a composed of two CDS coding for the E. guineensis DGAT-1-2 ( | + | This part is a composed of two CDS coding for the E. guineensis DGAT-1-2 (BBa_K2909003) and LPAAT-A (BBa_K2909002) enzymes, both tagged with a N-terminal HiBiT tag (BBa_K2909000).<br> |
Both enzymes are under the control of a nitrate-inducible Chlamydomonas reinhardtii promoter called NIT1. <br> | Both enzymes are under the control of a nitrate-inducible Chlamydomonas reinhardtii promoter called NIT1. <br> | ||
This part also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.<br><br> | This part also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.<br><br> | ||
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This part has been designed to enhance the overall production of triglycerides in Chlamydomonas reinhardtii. The N-terminal HiBiT tag allows a quick determination of the enzyme expression.<br><br> | This part has been designed to enhance the overall production of triglycerides in Chlamydomonas reinhardtii. The N-terminal HiBiT tag allows a quick determination of the enzyme expression.<br><br> | ||
| − | This part is a linear DNA that is the result of the digestion of the level M plasmid by the type IIS BsaI | + | 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. <br> |
It is this linear DNA that is integrated into the genomic DNA of Chlamydomonas reinhardtii during the transformation. | 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> | ||
| − | Bio | + | Bio(oil)gical Factory (iGEM Sorbonne Université 2019) |
=='''Characterization'''== | =='''Characterization'''== | ||
Latest revision as of 11:49, 4 September 2019
HiBiT-DGAT-1-2_HiBiT-LPAAT-A_HygroR E. guineensis
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3104
Illegal EcoRI site found at 5618
Illegal PstI site found at 2389
Illegal PstI site found at 4372 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3104
Illegal EcoRI site found at 5618
Illegal NheI site found at 5882
Illegal PstI site found at 2389
Illegal PstI site found at 4372
Illegal NotI site found at 217
Illegal NotI site found at 3316 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3104
Illegal EcoRI site found at 5618
Illegal BglII site found at 2297
Illegal BamHI site found at 2537
Illegal BamHI site found at 4214
Illegal BamHI site found at 5051
Illegal XhoI site found at 758
Illegal XhoI site found at 1020
Illegal XhoI site found at 3857 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3104
Illegal EcoRI site found at 5618
Illegal PstI site found at 2389
Illegal PstI site found at 4372 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3104
Illegal EcoRI site found at 5618
Illegal PstI site found at 2389
Illegal PstI site found at 4372
Illegal NgoMIV site found at 4927
Illegal NgoMIV site found at 6583
Illegal NgoMIV site found at 6765 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 2575
Illegal SapI.rc site found at 5089
Introduction
1- Biological background
This part is a composed of two CDS coding for the E. guineensis DGAT-1-2 (BBa_K2909003) and LPAAT-A (BBa_K2909002) enzymes, both tagged with a N-terminal HiBiT tag (BBa_K2909000).
Both enzymes are under the control of a nitrate-inducible Chlamydomonas reinhardtii promoter called NIT1.
This part also contains a Hygromycin resistance cassette called AphVII to select the recombinant clones.
This part has been designed to enhance the overall production of triglycerides in Chlamydomonas reinhardtii. The N-terminal HiBiT tag allows a quick determination of the enzyme expression.
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
- Schwinn, M. K. et al. CRISPR-Mediated Tagging of Endogenous Proteins with a Luminescent Peptide. ACS Chem. Biol. 13, 467–474 (2018).
- 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).
- 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).