Difference between revisions of "Part:BBa K389011:Design"

 
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K389011 short</partinfo>
 
<partinfo>BBa_K389011 short</partinfo>
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===Design Notes===
 
===Design Notes===
medium strong constitutive promotor, terminator to keep basal expression of kanamycin resistance low
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* medium strong constitutive promoter
 
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* The ''virG'' gene is mutated so it works without an ''rpoA'' subunit from ''Agrobacterium tumefaciens'' (YC Jung ''et al.'', 2004)
 
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* double terminator (forward) to keep expression of kanamycin resistance by the constitutive promoter low
 +
** double terminator <partinfo>B0017</partinfo> instead of <partinfo>B0015</partinfo> because of problems mentioned with <partinfo>B0012</partinfo>
 +
* Kanamycin resistance for screening with agar plates - the more the ''vir'' promoter is activated by VirG the more resistant the cells become to kanamycin.
  
 
===Source===
 
===Source===
  
synthetic ''virG'' gene, ''vir'' promotor from ''A. tumefaciens'' C58, parts.igem
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* ''virG'' gene synthesized by Mr. Gene (<partinfo>K389002</partinfo>)
 +
* ''vir'' promoter from ''A. tumefaciens'' C58 (<partinfo>K389003</partinfo>)
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* ''kanR'' gene made from BioBrick <partinfo>P1003</partinfo>
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* constitutive promoter and double terminator from parts.igem (<partinfo>J23110</partinfo>, <partinfo>B0017</partinfo>)
  
 
===References===
 
===References===
 +
YC Jung ''et al.'' (2004) Mutants of ''Agrobacterium tumefaciens virG'' Gene That Activate Transcription of ''vir'' Promoter in ''Escherichia coli'', ''Current Microbiol'' 49:334-340.

Latest revision as of 14:45, 16 October 2010

VirA screening device


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • medium strong constitutive promoter
  • The virG gene is mutated so it works without an rpoA subunit from Agrobacterium tumefaciens (YC Jung et al., 2004)
  • double terminator (forward) to keep expression of kanamycin resistance by the constitutive promoter low
  • Kanamycin resistance for screening with agar plates - the more the vir promoter is activated by VirG the more resistant the cells become to kanamycin.

Source

References

YC Jung et al. (2004) Mutants of Agrobacterium tumefaciens virG Gene That Activate Transcription of vir Promoter in Escherichia coli, Current Microbiol 49:334-340.