Difference between revisions of "Part:BBa K515100:Design"
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===Design Notes===
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===Design Notes===  
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1. designe RBS with RBS calculator - specify strength (site calculator)
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2. bc calculator takes into account the upstream region - added insulator which allows promoter switching.
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3. chose pVEG promoter bc it works in E. coli and B. sub. (developed program)
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4. codon optimisation steps
  
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An insulator sequence has been designed before each RBS to contain no homology with the vector, thereby avoiding recombination and allowing easier PCR removal of the individual parts from the device as well as promoter switching without influencing the RBS.  
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An insulator sequence has been designed upstream of the RBS (insert sequence). Its purpose is primarily to to contain no homology with the vector, thereby avoiding recombination and allowing easier PCR removal of the individual parts from the device as well as promoter switching without influencing the RBS.  
  
 
The coding regions have been optimised for B. subtilis and E. coli through the use of the program we have designed.
 
The coding regions have been optimised for B. subtilis and E. coli through the use of the program we have designed.
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Assembly
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1. sub parts designed and assembled by CPEC
 
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The parts were assembled into pSB1C3 by <a href="http://www.nature.com/nprot/journal/v6/n2/pdf/nprot.2010.181.pdf?WT.ec_id=NPROT-201102">CPEC assembly</a>.
 
The parts were assembled into pSB1C3 by <a href="http://www.nature.com/nprot/journal/v6/n2/pdf/nprot.2010.181.pdf?WT.ec_id=NPROT-201102">CPEC assembly</a>.

Revision as of 18:00, 13 September 2011

IAA biosynthetic genes under control of the Pveg2 promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 547
    Illegal BamHI site found at 1492
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 254
    Illegal NgoMIV site found at 2835
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

1. designe RBS with RBS calculator - specify strength (site calculator) 2. bc calculator takes into account the upstream region - added insulator which allows promoter switching. 3. chose pVEG promoter bc it works in E. coli and B. sub. (developed program) 4. codon optimisation steps

An insulator sequence has been designed upstream of the RBS (insert sequence). Its purpose is primarily to to contain no homology with the vector, thereby avoiding recombination and allowing easier PCR removal of the individual parts from the device as well as promoter switching without influencing the RBS.

The coding regions have been optimised for B. subtilis and E. coli through the use of the program we have designed.

Assembly 1. sub parts designed and assembled by CPEC The parts were assembled into pSB1C3 by CPEC assembly.

Source

Genomics sequence originating from P. savastanoi, synthesized by Eurofins.

References