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Part:BBa_K1413044:Design

Designed by: Sophia Belkhelfa   Group: iGEM14_Evry   (2014-10-15)
Revision as of 19:12, 1 November 2014 by SophiaB (Talk | contribs) (→‎Design Notes)

A fusion of Transposon Plasmid and pSB1C3


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 2415
    Illegal suffix found in sequence at 2437
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2415
    Illegal NheI site found at 947
    Illegal SpeI site found at 2438
    Illegal PstI site found at 2452
    Illegal NotI site found at 2421
    Illegal NotI site found at 2445
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2415
    Illegal BglII site found at 5
    Illegal BglII site found at 2628
    Illegal BamHI site found at 2475
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 2415
    Illegal suffix found in sequence at 2438
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 2415
    Illegal XbaI site found at 2430
    Illegal SpeI site found at 2438
    Illegal PstI site found at 2452
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This plasmid has been built by Golden Gate from pNK2 plasmid.





To isolate the two plasmids it would be possible to digeste the merged plasmid with BglII and then extract by gel eletrophoresis our Transposon plasmid, which only contains the core elements needed for the transposition. Between the two transposable elements iS10 we will eventually integrate the biobrick prefix and suffix to be able to integrate any biobrick in a genome.



Our aim was to merged pSB1C3 and pNK2 into a single plasmid, in order to have a plasmid that would replicate in non-pir cells. Also, our goal was to have a plasmid matching the requirements of the biobrick format. Here we used the pSB1C3 plasmid as the backbone. To isolate the two plasmids it would be possible to digeste the merged plasmid with BglII and then extract by gel eletrophoresis our Transposon plasmid, which only contains the core elements needed for the transposition. Between the two transposable elements iS10 we will eventually integrate the biobrick prefix and suffix to be able to integrate any biobrick in a genome.

Source

Each blend provide to plasmid pNK2. This is Bryan JESTER who give pNK2.

References