Difference between revisions of "Part:BBa K1744001:Design"

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<partinfo>BBa_K1744001 SequenceAndFeatures</partinfo>
 
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===Design Notes===
 
===Design Notes===
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The amilCP chromoprotein synthesis could be improved for the blue color to be visible in single copy faster. It has been of great concern through-out all the project and that is why we have put the strongest combination of promoter and RBS we have found to be well characterised. Still, expression in single copy is not so intense. In our case, amilCP can still be useful as a marker for plasmid background detection during recombineering experiment. Using only the kanamycin marker for recombineering, the blue colonies would represent plasmid transformants and not good recombinants. This use is good for us though, since background is a great concern in recombineering techniques, where screening can be long.
 
The amilCP chromoprotein synthesis could be improved for the blue color to be visible in single copy faster. It has been of great concern through-out all the project and that is why we have put the strongest combination of promoter and RBS we have found to be well characterised. Still, expression in single copy is not so intense. In our case, amilCP can still be useful as a marker for plasmid background detection during recombineering experiment. Using only the kanamycin marker for recombineering, the blue colonies would represent plasmid transformants and not good recombinants. This use is good for us though, since background is a great concern in recombineering techniques, where screening can be long.
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The part was constructed using a kanamycin resistance gene from plasmid pOK12 (available commercially) and amilCP from BBa_K592009. The promoter and RBS P1U8 were synthesized and come from Mutalik et al. (Nature 2013).
 
The part was constructed using a kanamycin resistance gene from plasmid pOK12 (available commercially) and amilCP from BBa_K592009. The promoter and RBS P1U8 were synthesized and come from Mutalik et al. (Nature 2013).
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===References===
 
===References===
 
Vivek K Mutalik, Joao C Guimaraes, Guillaume Cambray, Quynh-Anh Mai1, Marc Juul Christoffersen, Lance Martin, Ayumi Yu, Colin Lam, Cesar Rodriguez, Gaymon Bennett, Jay D Keasling, Drew Endy & Adam P Arkin, Quantitative estimation of activity and quality for collections of functional genetic elements, Nature methods, 10(4), 2013.
 
Vivek K Mutalik, Joao C Guimaraes, Guillaume Cambray, Quynh-Anh Mai1, Marc Juul Christoffersen, Lance Martin, Ayumi Yu, Colin Lam, Cesar Rodriguez, Gaymon Bennett, Jay D Keasling, Drew Endy & Adam P Arkin, Quantitative estimation of activity and quality for collections of functional genetic elements, Nature methods, 10(4), 2013.
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1744002 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K1744002 parameters</partinfo>
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Revision as of 07:04, 15 September 2015

P1U8-amilCP-kanR

Design Notes

The amilCP chromoprotein synthesis could be improved for the blue color to be visible in single copy faster. It has been of great concern through-out all the project and that is why we have put the strongest combination of promoter and RBS we have found to be well characterised. Still, expression in single copy is not so intense. In our case, amilCP can still be useful as a marker for plasmid background detection during recombineering experiment. Using only the kanamycin marker for recombineering, the blue colonies would represent plasmid transformants and not good recombinants. This use is good for us though, since background is a great concern in recombineering techniques, where screening can be long.


Sources

The part was constructed using a kanamycin resistance gene from plasmid pOK12 (available commercially) and amilCP from BBa_K592009. The promoter and RBS P1U8 were synthesized and come from Mutalik et al. (Nature 2013).


References

Vivek K Mutalik, Joao C Guimaraes, Guillaume Cambray, Quynh-Anh Mai1, Marc Juul Christoffersen, Lance Martin, Ayumi Yu, Colin Lam, Cesar Rodriguez, Gaymon Bennett, Jay D Keasling, Drew Endy & Adam P Arkin, Quantitative estimation of activity and quality for collections of functional genetic elements, Nature methods, 10(4), 2013.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1227
    Illegal NheI site found at 1630
    Illegal NheI site found at 1653
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1166
    Illegal BamHI site found at 1613
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1001
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 983