Difference between revisions of "Part:BBa K590017"
 
Line 8: Line 8:
  
 
[[Image:IGEM11 HIEJKL partregistry.png|500px|center]]
 
[[Image:IGEM11 HIEJKL partregistry.png|500px|center]]
−
 
−
The gel image on the left is HIEJKL(~9000bp), we extracted this band and "gibsoned" it with pGA3k3 plasmid backbone(~3000bp), the right image.
 
−
 
−
[[Image:Igem2011 HIEJKL gel.png|thumb|left|1kb Ladder (left), mamHIEJKL (right)]][[Image:Igem2011 3k3 gel.png|thumb|left|1kb Ladder (left), 3k3 backbone (right)]]
 
−
 
−
 
  
  

Latest revision as of 23:18, 28 September 2011

mamHIEJKL in vector pGA3K3

This part is made by [http://2011.igem.org/Team:Washington UW iGEM Team 2011]. It includes 6 genes mamHI, mamE, mamJ, mamKL as well as the native mamAB promoter, all in a pGA3k3 plasmid backbone. All the genes were found essential for magnetosome formation in Magnetospirillum magneticum strain AMB-1.

Usage and Biology

We made this construct from 2 kilobase gene extractions from AMB-1. These fragments were assembled using the [http://2010.igem.org/Team:Cambridge/Gibson/RFC Gibson cloning] method instead of standard assembly. This super-assembly is known as "Part one" of the full 16kb mamAB reassembly project. This part is in a medium copy backbone and it belongs to the [http://2011.igem.org/Team:Washington/Magnetosomes/Magnet_Toolkit Magnetosome Toolkit].

IGEM11 HIEJKL partregistry.png











After Gibson cloning and transformation, cell chain formation was observed in microscopy(below) when this plasmid was transformed in E.coli.

HIEJKL 1b wiki.png


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 5175
    Illegal PstI site found at 8836
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 5175
    Illegal PstI site found at 8836
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 8540
    Illegal XhoI site found at 89
    Illegal XhoI site found at 6375
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 5175
    Illegal PstI site found at 8836
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 5175
    Illegal PstI site found at 8836
    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 6754
    Illegal NgoMIV site found at 8562
    Illegal NgoMIV site found at 8929
    Illegal AgeI site found at 5849
    Illegal AgeI site found at 6264
  • 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 6947
    Illegal SapI site found at 825
    Illegal SapI site found at 2160
    Illegal SapI site found at 3491
    Illegal SapI.rc site found at 9208
    Illegal SapI.rc site found at 9529