Difference between revisions of "Part:BBa K2483004:Design"

 
(→‎Source)
 
Line 16: Line 16:
 
For the sources for dCas9 and the IAA fusion proteins, see their respective parts (BBa_K2483002 and BBa_K2483000).  
 
For the sources for dCas9 and the IAA fusion proteins, see their respective parts (BBa_K2483002 and BBa_K2483000).  
 
The sgRNA design was taken from iGEM Warwick and modified to include the target sites we were using (BBa_K1994017 and BBa_K1994016).
 
The sgRNA design was taken from iGEM Warwick and modified to include the target sites we were using (BBa_K1994017 and BBa_K1994016).
 +
 +
The promoter and terminator for the sgRNAs were taken from this paper:
 +
Zalatan, J. G., Lee, M. E., Almeida, R., Gilbert, L. A., Whitehead, E. H., La Russa, M., et al. (2015). Engineering complex synthetic transcriptional programs with CRISPR RNA scaffolds. Cell 160, 339–350. doi:10.1016/j.cell.2014.11.052.
  
 
===References===
 
===References===

Latest revision as of 13:28, 16 September 2017


regulated dCas9 with sgRNAs and IAA enzymes fused to MS2 and PP7


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3004
    Illegal NheI site found at 9656
    Illegal NheI site found at 9930
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 4412
    Illegal BglII site found at 6221
    Illegal BglII site found at 9644
    Illegal BglII site found at 9918
    Illegal BamHI site found at 725
    Illegal BamHI site found at 7166
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 5928
    Illegal NgoMIV site found at 8872
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The LacI-dCas9 construct was put in reverse complement to the other parts to utilize the temrinators on both ends of the plasmid backbone, thereby reducing additional planning.



Source

For the sources for dCas9 and the IAA fusion proteins, see their respective parts (BBa_K2483002 and BBa_K2483000). The sgRNA design was taken from iGEM Warwick and modified to include the target sites we were using (BBa_K1994017 and BBa_K1994016).

The promoter and terminator for the sgRNAs were taken from this paper: Zalatan, J. G., Lee, M. E., Almeida, R., Gilbert, L. A., Whitehead, E. H., La Russa, M., et al. (2015). Engineering complex synthetic transcriptional programs with CRISPR RNA scaffolds. Cell 160, 339–350. doi:10.1016/j.cell.2014.11.052.

References