Difference between revisions of "Part:BBa K2909002:Design"
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===Design Notes=== | ===Design Notes=== | ||
The coding sequence was first codon optimized for C. reinhardtii by reverse translating its protein sequence using a software that takes into account the codon usage of C. reinhardtii.<br> | The coding sequence was first codon optimized for C. reinhardtii by reverse translating its protein sequence using a software that takes into account the codon usage of C. reinhardtii.<br> | ||
| − | The sequence was then standardized to fit the B3-B4 position of the C. reinhardtii MoClo Kit by adding BbsI recognition sites and fusion sites on both ends.<br> | + | The sequence was then standardized in the Phytobrick standard to fit the B3-B4 position of the C. reinhardtii MoClo Kit by adding BbsI recognition sites and fusion sites on both ends.<br> |
The stop codon was removed to allow for tagging at the C-terminal position, the stop codon being provided by the part on the position B5. | The stop codon was removed to allow for tagging at the C-terminal position, the stop codon being provided by the part on the position B5. | ||
| + | |||
===Source=== | ===Source=== | ||
Latest revision as of 08:36, 30 August 2019
LPAAT-A-B3-B4 E. guineensis codon optimized for C. reinhardtii
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 474
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 474
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 316
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 474
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 474
Illegal NgoMIV site found at 204
Illegal NgoMIV site found at 1029 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The coding sequence was first codon optimized for C. reinhardtii by reverse translating its protein sequence using a software that takes into account the codon usage of C. reinhardtii.
The sequence was then standardized in the Phytobrick standard to fit the B3-B4 position of the C. reinhardtii MoClo Kit by adding BbsI recognition sites and fusion sites on both ends.
The stop codon was removed to allow for tagging at the C-terminal position, the stop codon being provided by the part on the position B5.
Source
Predicted sequence from the E. guineensis genome sequencing.
NCBI Reference Sequence: XM_010908457.2
https://www.ncbi.nlm.nih.gov/nuccore/1130625520
References
- Dussert, S. et al. Comparative Transcriptome Analysis of Three Oil Palm Fruit and Seed Tissues That Differ in Oil Content and Fatty Acid Composition. PLANT PHYSIOLOGY 162, 1337–1358 (2013).
- 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).