Difference between revisions of "Part:BBa K590063"
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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]. | 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]. | ||
− | + | This part is contributed to the [http://2011.igem.org/Team:Washington/Magnetosomes/Magnet_Toolkit Magnetosome toolkit], it also serves as an evident that Gibson Cloning method is capable for combining multiple inserts. | |
===Usage and Biology=== | ===Usage and Biology=== |
Revision as of 07:15, 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.
This part is contributed to the [http://2011.igem.org/Team:Washington/Magnetosomes/Magnet_Toolkit Magnetosome toolkit], it also serves as an evident that Gibson Cloning method is capable for combining multiple inserts.
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.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 12370
Illegal PstI site found at 5183
Illegal PstI site found at 8860 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 12370
Illegal PstI site found at 5183
Illegal PstI site found at 8860 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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