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

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===Design Notes===
 
===Design Notes===
* medium strong constitutive promoter,  
+
* medium strong constitutive promoter
* double terminator (forward) to keep basal expression of kanamycin resistance low
+
* 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
 
** double terminator <partinfo>B0017</partinfo> instead of <partinfo>B0015</partinfo> because of problems mentioned with <partinfo>B0012</partinfo>
 
** double terminator <partinfo>B0017</partinfo> instead of <partinfo>B0015</partinfo> because of problems mentioned with <partinfo>B0012</partinfo>
* kanamycin resistance for screening with plates
+
* 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===
  
 
* ''virG'' gene synthesized by Mr. Gene (<partinfo>K389002</partinfo>)
 
* ''virG'' gene synthesized by Mr. Gene (<partinfo>K389002</partinfo>)
* ''vir'' promoter from ''A. tumefaciens'' C58 (partinfo>K389003</partinfo>)
+
* ''vir'' promoter from ''A. tumefaciens'' C58 (<partinfo>K389003</partinfo>)
 +
* ''kanR'' gene made from BioBrick <partinfo>P1003</partinfo>
 
* constitutive promoter and double terminator from parts.igem (<partinfo>J23110</partinfo>, <partinfo>B0017</partinfo>)
 
* 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.