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Part:BBa_K1413044:Design

Designed by: Sophia Belkhelfa   Group: iGEM14_Evry   (2014-10-15)
Revision as of 20:49, 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 was built by Golden Gate from pNK2 plasmid.


Fig. 1 Map of pNK2 plasmid with restrictions sites E X S P



The biobrick must be digested with bglII, extracted and ligated to form the transposon plasmid. To integrate any DNA sequence/biobrick into the genome, biobrick prefix and suffix can be used. This will integrate the DNA between the two transposable elements IS10.


Fig. 2 Map of transposon plasmid
The goal was to merge pSB1C3 and pNK2 into a single plasmid, in order to have a plasmid that would replicate in non-pir cells, and matching the requirements of the biobrick format. The pSB1C3 plasmid was used as backbone.

Source

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

References