Difference between revisions of "Part:BBa K510045:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K510045 short</partinfo>
 
<partinfo>BBa_K510045 short</partinfo>
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===Design Notes===
 
===Design Notes===
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The BBa_K510019 BioBrick was inserted within the BCS of pUC18Sfi-miniTn7BB-Gm (BBa K510000) by EcoRI and PstI clonning. In order to construct the pUC18Sfi-miniTn7BB-Gm delivery plasmid, the miniTn7BB-Gm transposon was cleaved from the commercial plasmid pMA (Mr. Gene) with SfiI and ligated to SfiI-digested pUC18Sfi. This strategy removes all multi-cloning restriction sites from pUC18Sfi, except for the flanking SfiI, thus guaranteeing that the unique sites in the transposon are not duplicated. The remove of the multi-cloning restriction sites was verified by analytic digestions.
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The [https://parts.igem.org/Part:BBa_K510019 BBa_K510019] BioBrick was inserted within the BCS of pUC18Sfi-miniTn7BB-Gm ([https://parts.igem.org/Part:BBa_K510000 BBa K510000]) by EcoRI and PstI clonning. In order to construct the pUC18Sfi-miniTn7BB-Gm delivery plasmid, the miniTn7BB-Gm transposon was cleaved from the commercial plasmid pMA (Mr. Gene) with SfiI and ligated to SfiI-digested pUC18Sfi. This strategy removes all multi-cloning restriction sites from pUC18Sfi, except for the flanking SfiI, thus guaranteeing that the unique sites in the transposon are not duplicated. The remove of the multi-cloning restriction sites was verified by analytic digestions.
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===Source===
 
===Source===
  
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The miniTn7BB-Gm minitransposon was synthesized commercially and pUC18SfiI is a commercial vector.
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The miniTn7BB-Gm minitransposon and the improved flip-flop (module I) were synthesized commercially and pUC18SfiI is a commercial vector.
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The improved flip-flop (module I) was synthesized commercially.
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===References===
 
===References===

Latest revision as of 11:08, 22 October 2011

pUC18Sfi-miniTn7BB-Gm-improved_flipflop_(module I)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4367
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4367
    Illegal NheI site found at 7526
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 4373
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4367
    Illegal BglII site found at 3051
    Illegal BglII site found at 3322
    Illegal BglII site found at 3608
    Illegal BamHI site found at 5336
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4367
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4367
    Illegal XbaI site found at 4382
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal AgeI site found at 8788
    Illegal AgeI site found at 8900
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1681
    Illegal BsaI site found at 4667
    Illegal SapI.rc site found at 2763


Design Notes

The BBa_K510019 BioBrick was inserted within the BCS of pUC18Sfi-miniTn7BB-Gm (BBa K510000) by EcoRI and PstI clonning. In order to construct the pUC18Sfi-miniTn7BB-Gm delivery plasmid, the miniTn7BB-Gm transposon was cleaved from the commercial plasmid pMA (Mr. Gene) with SfiI and ligated to SfiI-digested pUC18Sfi. This strategy removes all multi-cloning restriction sites from pUC18Sfi, except for the flanking SfiI, thus guaranteeing that the unique sites in the transposon are not duplicated. The remove of the multi-cloning restriction sites was verified by analytic digestions.

Source

The miniTn7BB-Gm minitransposon and the improved flip-flop (module I) were synthesized commercially and pUC18SfiI is a commercial vector.

References