Difference between revisions of "Part:BBa K2909014"
Laurent CHEN (Talk | contribs) |
Laurent CHEN (Talk | contribs) (→Introduction) |
||
| Line 11: | Line 11: | ||
This part is a composed of two CDS coding for the E. guineensis DGAT-1-2 (BBa_K2909002) and LPAAT-A (BBa_K2909003)enzymes , both tagged with a C-terminal HiBiT tag (BBa_K2909001).<br> | This part is a composed of two CDS coding for the E. guineensis DGAT-1-2 (BBa_K2909002) and LPAAT-A (BBa_K2909003)enzymes , both tagged with a C-terminal HiBiT tag (BBa_K2909001).<br> | ||
| − | Both enzymes are under the control of | + | Both enzymes are under the control of an ammonium-controlled 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> | ||
This part has been designed to enhance the overall production of triglycerides in Chlamydomonas reinhardtii. The C-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 C-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'''== | ||
Revision as of 08:37, 29 August 2019
DGAT-1-2-HiBiT_LPAAT-A-HiBiT_HygroR E. guineensis
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3079
Illegal EcoRI site found at 5568
Illegal PstI site found at 2349
Illegal PstI site found at 4307 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3079
Illegal EcoRI site found at 5568
Illegal NheI site found at 5832
Illegal PstI site found at 2349
Illegal PstI site found at 4307
Illegal NotI site found at 217
Illegal NotI site found at 3291 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3079
Illegal EcoRI site found at 5568
Illegal BglII site found at 2257
Illegal BamHI site found at 4149
Illegal XhoI site found at 758
Illegal XhoI site found at 980
Illegal XhoI site found at 3832 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3079
Illegal EcoRI site found at 5568
Illegal PstI site found at 2349
Illegal PstI site found at 4307 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 5
Illegal EcoRI site found at 3079
Illegal EcoRI site found at 5568
Illegal PstI site found at 2349
Illegal PstI site found at 4307
Illegal NgoMIV site found at 4862
Illegal NgoMIV site found at 6533
Illegal NgoMIV site found at 6715 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 2550
Illegal SapI.rc site found at 5039
Introduction
1- Biological background
This part is a composed of two CDS coding for the E. guineensis DGAT-1-2 (BBa_K2909002) and LPAAT-A (BBa_K2909003)enzymes , both tagged with a C-terminal HiBiT tag (BBa_K2909001).
Both enzymes are under the control of an ammonium-controlled 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 C-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).