Difference between revisions of "Part:BBa K615000:Design"

 
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===Source===
 
===Source===
  
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EcNR2 E. coli strain
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Base strain was EcNR2, a modified E. coli strain as described in Wang et al (2009).
 +
 
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URA3 and His3 are genes from yeast.
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References:
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Harris H. Wang, Farren J. Isaacs, Peter A. Carr, Zachary Z. Sun, George Xu, Craig R. Forest, George M. Church. Programming cells by multiplex genome engineering and accelerated evolution. (2009). Nature, 460(7257):894-8.
  
 
===References===
 
===References===

Revision as of 21:26, 21 September 2011

E. coli strain for His3-URA3 one-hybrid selection system


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
    Illegal PstI site found at 1762
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
    Illegal PstI site found at 1762
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 845
    Illegal BglII site found at 965
    Illegal BglII site found at 1478
    Illegal BglII site found at 1538
    Illegal BamHI site found at 1768
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
    Illegal PstI site found at 1762
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 632
    Illegal PstI site found at 952
    Illegal PstI site found at 1762
    Illegal NgoMIV site found at 181
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2458
    Illegal SapI site found at 121
    Illegal SapI site found at 331
    Illegal SapI.rc site found at 2305


Design Notes

HisB and PyrF were knocked out using multiplex automated genome engineering (MAGE). Lambda red was employed to replace rpoZ with a zeocin cassette and to insert the kanamycin cassette-Zif268 binding site-His3-URA3 construct.


Source

Base strain was EcNR2, a modified E. coli strain as described in Wang et al (2009).

URA3 and His3 are genes from yeast.

References: Harris H. Wang, Farren J. Isaacs, Peter A. Carr, Zachary Z. Sun, George Xu, Craig R. Forest, George M. Church. Programming cells by multiplex genome engineering and accelerated evolution. (2009). Nature, 460(7257):894-8.

References