Device

Part:BBa_K1088013

Designed by: Andreas Kjr   Group: iGEM13_SDU-Denmark   (2013-08-28)

B. subtilis dxs (lac promoter with lac inhibitor: IPTG inducible)

This part consist of the dxs gene derived from B. subtilis under the control of the lac promoter and has a strong RBS.

To repress expression from the lac promoter, the lacI:LVA gene (under a constitutive promoter with a strong RBS and a efficient terminator) was placed upstream to the dxs gene. The repression can be relieved with addition of IPTG, which binds and inhibits the function of LacI:LVA.

The levels of protein expression were measured with a similar brick with linker-GFP fused C-terminal to Dxs (BBa_1088009) in E. coli K-12 MG1655 using fluorescence activated cell sorting (FACS). The experiment proved that the reporter fusion wasn't expressed when the strain was grown without IPTG in the media, and that the expression was at its maximum approximately 90 min after induction with 1 mM IPTG. See BBa_1088009 for more details.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2505
    Illegal EcoRI site found at 3162
    Illegal PstI site found at 2563
    Illegal PstI site found at 3009
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2505
    Illegal EcoRI site found at 3162
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal PstI site found at 2563
    Illegal PstI site found at 3009
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2505
    Illegal EcoRI site found at 3162
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2505
    Illegal EcoRI site found at 3162
    Illegal PstI site found at 2563
    Illegal PstI site found at 3009
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2505
    Illegal EcoRI site found at 3162
    Illegal PstI site found at 2563
    Illegal PstI site found at 3009
    Illegal NgoMIV site found at 2462
    Illegal AgeI site found at 2355
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2304
    Illegal SapI.rc site found at 3003


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