Part:BBa_K3781003:Design
mVenus, MoCloMania B3_B4
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 202
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 202
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 202
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 202
Illegal NgoMIV site found at 679 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part was adapted from a Chlamy-optimized plasmid, in which it contained a RBCS2 intron, we had to design special primers that would render the genetic regions upstream and downstream of the intron as individual parts via PCR. These then could be re-assembled into an intact mVenus gene with the help of invertedly oriented BbsI restriction sites introduced with the primers. Due to the high resemblence in codon usage between Chlamydomonas reinhardtii and Leishmania tarentolae, the codons were not further optimized.
Source
The sequence was directly derived from the plasmid pCM0-086 of the MoClo Toolkit distributed by Crozet et al.[1] The protein is a mutated version of the yellow fluorescent protein EYFP which in itself is derived via mutagenesis from the green fluorescent protein avGFP isolated from the jellyfish Aequorea victoria.[2] mVenus's amino acid sequence can be found in the NCBI GenBank under AAZ65844.1
References
- ↑ Crozet et al. (2018) Birth of a Photosynthetic Chassis: A MoClo Toolkit Enabling Synthetic Biology in the Microalga Chlamydomonas reinhardtii. ACS Synthetic Biology 7:2074–2086
- ↑ M. Ormo, A. B. Cubitt, K. Kallio, L. A. Gross, R. Y. Tsien, S. J. Remington, Crystal structure of the Aequorea victoria green fluorescent protein. Science 273, 1392–1395 (1996)