Difference between revisions of "Part:BBa K1088011"
 
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<partinfo>BBa_K1088011 short</partinfo>
 
<partinfo>BBa_K1088011 short</partinfo>
  
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This part has the B. sub dxs (BBa_K1088000) controlled by the common Lac promoter. Notice that there overexpression of LacI is required for regulation of the Lac promoter.
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This part contains the ''dxs'' gene derived from ''B. subtilis'' ([https://parts.igem.org/Part:BBa_K1088000 BBa_K1088000]) controlled by the ''lac'' promoter and has a strong RBS. The part was build to increase the flow through the MEP (methylerythritol phosphate) pathway, and to test whether Dxs derived from ''E. coli'' or ''B. subtilis'' increased the flow through the MEP pathway the most in ''E. coli''.
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The ''lac'' promoter is repressed by the LacI protein, and repression can be relived through allosteric binding of IPTG to the repressor.
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Fluorescence activated cell sorting (FACS) was used to assay the protein expression profile of a similar part ([https://parts.igem.org/Part:BBa_K1088008 Ba_K1088008]) with linker-GFP attached C-terminal to Dxs. Experiments proved that this part is constitutively active when LacI isn't overexpressed in the cell. See [https://parts.igem.org/Part:BBa_K1088008 Ba_K1088008] for more details.  
  
  

Latest revision as of 18:23, 28 October 2013

B. subtilis dxs (lac promoter without lac inhibitor)

This part contains the dxs gene derived from B. subtilis (BBa_K1088000) controlled by the lac promoter and has a strong RBS. The part was build to increase the flow through the MEP (methylerythritol phosphate) pathway, and to test whether Dxs derived from E. coli or B. subtilis increased the flow through the MEP pathway the most in E. coli.

The lac promoter is repressed by the LacI protein, and repression can be relived through allosteric binding of IPTG to the repressor.

Fluorescence activated cell sorting (FACS) was used to assay the protein expression profile of a similar part (Ba_K1088008) with linker-GFP attached C-terminal to Dxs. Experiments proved that this part is constitutively active when LacI isn't overexpressed in the cell. See Ba_K1088008 for more details.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1279
    Illegal EcoRI site found at 1936
    Illegal PstI site found at 1337
    Illegal PstI site found at 1783
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1279
    Illegal EcoRI site found at 1936
    Illegal PstI site found at 1337
    Illegal PstI site found at 1783
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1279
    Illegal EcoRI site found at 1936
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1279
    Illegal EcoRI site found at 1936
    Illegal PstI site found at 1337
    Illegal PstI site found at 1783
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1279
    Illegal EcoRI site found at 1936
    Illegal PstI site found at 1337
    Illegal PstI site found at 1783
    Illegal NgoMIV site found at 1236
    Illegal AgeI site found at 1129
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1078
    Illegal SapI.rc site found at 1777