Difference between revisions of "Part:BBa K2560261:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | The part contains the | + | The part contains the 3' part of the <i>mcr</i> gene of <i>C. aurantiacus</i> (AY530019). This region is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011). |
It was shown that a splitted version of Mcr with separated C- and N-terminal domains increases enzyme activity (Liu et al., 2013). If these splitted genes shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560261 BBa_K2560261] and this part. If the gene for the whole enzyme shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560260 BBa_K2560260] | It was shown that a splitted version of Mcr with separated C- and N-terminal domains increases enzyme activity (Liu et al., 2013). If these splitted genes shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560261 BBa_K2560261] and this part. If the gene for the whole enzyme shall be used take [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560260 BBa_K2560260] | ||
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The sequence was codonoptimized for <i>V. natriegens</i> ATCC 14048. According to Liu et al, we designed the part so that McrN contains amino acid 1-549 of the overall protein and McrC amino acid 550-1219 (Liu et al., 2013). | The sequence was codonoptimized for <i>V. natriegens</i> ATCC 14048. According to Liu et al, we designed the part so that McrN contains amino acid 1-549 of the overall protein and McrC amino acid 550-1219 (Liu et al., 2013). | ||
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===Source=== | ===Source=== | ||
Revision as of 14:19, 30 September 2018
mcrC - C-terminal domain of Malonyl-CoA Reductase from Chloroflexus aurantiacus
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 288
Illegal NheI site found at 1690 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 361
Illegal NgoMIV site found at 1975
Illegal AgeI site found at 1524 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The part contains the 3' part of the mcr gene of C. aurantiacus (AY530019). This region is flanked by overhangs which are Phytobrick- and MoClo-compatible and by two BsaI recognition sites (Weber et al., 2011).
It was shown that a splitted version of Mcr with separated C- and N-terminal domains increases enzyme activity (Liu et al., 2013). If these splitted genes shall be used take BBa_K2560261 and this part. If the gene for the whole enzyme shall be used take BBa_K2560260
GGTCTCGAATG-coding_region-GCTTTGAGACC
The sequence was codonoptimized for V. natriegens ATCC 14048. According to Liu et al, we designed the part so that McrN contains amino acid 1-549 of the overall protein and McrC amino acid 550-1219 (Liu et al., 2013).
Source
Source of the part:
Genome: Chloroflexus aurantiacus strain OK-70-fl
Accession number of gene: AY530019, amino acid 550-1219
Accession number of encoded protein: AAS20429
mcrC was codonoptimized for V. natriegens and then synthetisized and integrated into the vector BBa_K2560002 via BsmBI
References
Liu, C., Wang, Q., Xian, M., Ding, Y., Zhao, G., 2013. Dissection of Malonyl-Coenzyme A Reductase of Chloroflexus aurantiacus Results in Enzyme Activity Improvement. PLoS One 8, 1–8. https://doi.org/10.1371/journal.pone.0075554
Strauss, G., Fuchs, G., 1993. Enzymes of a novel autotrophic CO2 fixation pathway in the phototrophic bacterium Chloroflexus aurantiacus, the 3-hydroxypropionate cycle. Eur. J. Biochem. 215, 633–43.
Weber, E., Engler, C., Gruetzner, R., Werner, S., Marillonnet, S., 2011. A Modular Cloning System for Standardized Assembly of Multigene Constructs. PLoS One 6, e16765. https://doi.org/10.1371/journal.pone.0016765