Difference between revisions of "Part:BBa K2560262:Design"

 
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===Design Notes===
 
===Design Notes===
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The part contains the 5' 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).
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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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GGTCTCG<b>AATG</b>-coding_region-<b>GCTT</b>TGAGACC
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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).
  
  
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===Source===
 
===Source===
  
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egfqegqeg
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Source of the part:
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Genome: <i>Chloroflexus aurantiacus</i> strain OK-70-fl
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Accession number of gene: AY530019, amino acid 1-549
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Accession number of encoded protein: AAS20429
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<i>mcrN</i> was codonoptimized for <i>V. natriegens</i> and then synthetisized and integrated into the vector [https://parts.igem.org/wiki/index.php?title=Part:BBa_K2560002 BBa_K2560002] via BsmBI
  
 
===References===
 
===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
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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

Revision as of 14:24, 30 September 2018


mcrN - N-terminal domain of Malonyl-CoA Reductase from Chloroflexus aurantiacus


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 885
    Illegal BamHI site found at 213
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1188
    Illegal AgeI site found at 1270
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The part contains the 5' 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 1-549

Accession number of encoded protein: AAS20429

mcrN 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