Difference between revisions of "Part:BBa K1413044:Design"

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===Design Notes===
 
===Design Notes===
 
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This plasmid was built by Golden Gate from pNK2 plasmid.  
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This BioBrick was built by Golden Gate from pNK2 plasmid.  
  
 
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<img src="https://static.igem.org/mediawiki/parts/6/64/PNK2.png" width=50%/>
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<div align="center"><img src="https://static.igem.org/mediawiki/parts/6/64/PNK2.png" width=50%/></div>
 
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<b> Fig. 1</b> Map of pNK2 plasmid with restrictions sites E X S P
 
<b> Fig. 1</b> Map of pNK2 plasmid with restrictions sites E X S P
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<br/>To isolate the two plasmids it would be possible to digest 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.
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<br/>The biobrick must be digested with bglII, extracted and ligated to form the transposon plasmid.  
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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.  
  
 
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<img src="https://static.igem.org/mediawiki/parts/b/b4/Plqs%3Bid.png" width=30%/>
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<div align="center"><img src="https://static.igem.org/mediawiki/parts/b/b4/Plqs%3Bid.png" width=30%/></div>
 
<br/><b> Fig. 2</b> Map of transposon plasmid
 
<br/><b> Fig. 2</b> Map of transposon plasmid
 
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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.
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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.
−
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.
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===Source===
 
===Source===
  
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Each blend provide to plasmid pNK2. This is Bryan JESTER who give pNK2.  
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Each blend provide to plasmid pNK2. This plasmid was given by a member of the institute of systems and synthetic biology (Evry, France), the researcher Brian Jester.
  
 
===References===
 
===References===

Latest revision as of 18:03, 2 November 2014

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 BioBrick 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 plasmid was given by a member of the institute of systems and synthetic biology (Evry, France), the researcher Brian Jester.

References