Difference between revisions of "Part:BBa K590063"

 
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__NOTOC__
 
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<partinfo>BBa_K590063 short</partinfo>
 
<partinfo>BBa_K590063 short</partinfo>
  
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mamH, I, E, J, K, L all in one vector
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This part is made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011] using Gibson cloning method. It includes mamH, I, E, J, K, L all in [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590014 pGA3k3_placGFP].
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<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
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We first extracted [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590000 mamHI], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590001 mamE], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590002 mamJ], [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590003 mamKL] from the ''Magnetospirillum magneticum'' strain AMB-1, then we used Gibson cloning method to fuse the 4 groups of genes together to make a big HIEJKL insert. Followed by another gibson cloning reaction to combine the insert with the plasmid backbone [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590014 pGA3k3_placGFP].
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Below is the gel image of HIEJKL(~9000bp), we extracted this band and "gibsoned" it with [https://parts.igem.org/wiki/index.php?title=Part:BBa_K590014 pGA3k3_placGFP] plasmid backbone.
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[[Image:Igem2011 HIEJKL gel.png|thumb|center|1kb Ladder (left), mamHIEJKL (right)]]
  
 
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Revision as of 07:12, 22 September 2011

mamHIEJKL_pLacGFP_pGA3K3

This part is made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011] using Gibson cloning method. It includes mamH, I, E, J, K, L all in pGA3k3_placGFP.


Usage and Biology

We first extracted mamHI, mamE, mamJ, mamKL from the Magnetospirillum magneticum strain AMB-1, then we used Gibson cloning method to fuse the 4 groups of genes together to make a big HIEJKL insert. Followed by another gibson cloning reaction to combine the insert with the plasmid backbone pGA3k3_placGFP.

Below is the gel image of HIEJKL(~9000bp), we extracted this band and "gibsoned" it with pGA3k3_placGFP plasmid backbone.

1kb Ladder (left), mamHIEJKL (right)

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 12370
    Illegal PstI site found at 5183
    Illegal PstI site found at 8860
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 12370
    Illegal NheI site found at 11025
    Illegal PstI site found at 5183
    Illegal PstI site found at 8860
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 12370
    Illegal BglII site found at 8564
    Illegal BglII site found at 12385
    Illegal BamHI site found at 9642
    Illegal XhoI site found at 89
    Illegal XhoI site found at 6383
    Illegal XhoI site found at 9657
    Illegal XhoI site found at 9818
    Illegal XhoI site found at 10661
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 12370
    Illegal PstI site found at 5183
    Illegal PstI site found at 8860
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 12370
    Illegal PstI site found at 5183
    Illegal PstI site found at 8860
    Illegal NgoMIV site found at 238
    Illegal NgoMIV site found at 1254
    Illegal NgoMIV site found at 2589
    Illegal NgoMIV site found at 3920
    Illegal NgoMIV site found at 6770
    Illegal NgoMIV site found at 8586
    Illegal NgoMIV site found at 8953
    Illegal AgeI site found at 5857
    Illegal AgeI site found at 6272
    Illegal AgeI site found at 11111
    Illegal AgeI site found at 11434
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 651
    Illegal BsaI site found at 1986
    Illegal BsaI site found at 3317
    Illegal BsaI.rc site found at 6963
    Illegal BsaI.rc site found at 12139
    Illegal SapI site found at 825
    Illegal SapI site found at 2160
    Illegal SapI site found at 3491
    Illegal SapI.rc site found at 9232
    Illegal SapI.rc site found at 9553