Difference between revisions of "Part:BBa K2909010"
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<h3> 1- Biological background </h3> | <h3> 1- Biological background </h3> | ||
| − | This part is a CDS coding for the E. guineensis LPAAT-A enzyme tagged with a C-terminal HiBiT tag under the control of | + | This part is a CDS coding for the E. guineensis LPAAT-A enzyme tagged with a C-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> |
| − | This part has been designed to characterize the expression of the E. guineensis LPAAT-A enzyme that we adapted for C. reinhardtii (BBa_K2909002) | + | 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 C-terminal HibiT tag (BBa_K2909001).<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. 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 08:21, 29 August 2019
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 2494
Illegal PstI site found at 1233 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 5
Illegal EcoRI site found at 2494
Illegal NheI site found at 2758
Illegal PstI site found at 1233
Illegal NotI site found at 217 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 5
Illegal EcoRI site found at 2494
Illegal BamHI site found at 1075
Illegal XhoI site found at 758 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 5
Illegal EcoRI site found at 2494
Illegal PstI site found at 1233 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 5
Illegal EcoRI site found at 2494
Illegal PstI site found at 1233
Illegal NgoMIV site found at 1788
Illegal NgoMIV site found at 3459
Illegal NgoMIV site found at 3641 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1965
Introduction
1- Biological background
This part is a CDS coding for the E. guineensis LPAAT-A enzyme tagged with a C-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 C-terminal HibiT tag (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
- 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).