Difference between revisions of "Part:BBa K2148001:Design"

 
 
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===Design Notes===
 
===Design Notes===
 
Added phytobrick fusion sites
 
Added phytobrick fusion sites
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This part along with our other antibiotic resistance part BBa_K2148002 (aphA6- resistance to kanamycin) allows for the implementation of co-transformation experiments in chloroplasts.
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In our experimental strategy we aimed to introduce CRISPR/CAS9 technology into <i>C. reinhardtii</i> to aid in the process of achieving homoplasmy in a quicker way, major bottleneck in this system, by site-directed integration of the gene of interest. We addressed the potential issue of biological containment by placing cas9 on an independent plasmid (the "driver"- see image  below) linked to a different antibiotic to that of the gene of interest (on the "CoI" plasmid) so that the selective pressure of the driver could be removed (by removing the antibiotic from the medium) and eventually lost from the strain. For more details please check out our wiki: http://2016.igem.org/Team:Cambridge-JIC/Design       
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<b> INSERT DIAGRAM HERE </b>   
  
  

Latest revision as of 16:43, 13 October 2016


aadA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 652
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Added phytobrick fusion sites

This part along with our other antibiotic resistance part BBa_K2148002 (aphA6- resistance to kanamycin) allows for the implementation of co-transformation experiments in chloroplasts.

In our experimental strategy we aimed to introduce CRISPR/CAS9 technology into C. reinhardtii to aid in the process of achieving homoplasmy in a quicker way, major bottleneck in this system, by site-directed integration of the gene of interest. We addressed the potential issue of biological containment by placing cas9 on an independent plasmid (the "driver"- see image below) linked to a different antibiotic to that of the gene of interest (on the "CoI" plasmid) so that the selective pressure of the driver could be removed (by removing the antibiotic from the medium) and eventually lost from the strain. For more details please check out our wiki: http://2016.igem.org/Team:Cambridge-JIC/Design


INSERT DIAGRAM HERE


Source

pUC-atpX-AAD

References