Plasmid

Part:BBa_K1422505

Designed by: Casey Martin   Group: iGEM14_Purdue   (2014-10-17)


Lactose-inducible killswitch, produces SAMS and NAS1

This plasmid includes a killswitch and produces SAMS (BBa_K1422500) and NAS1 (BBa_K1422501), two of the four enzymes used in the production of corn phytosiderophores. The killswitch (BBa_K1422504) is lactose and IPTG inducible and uses the antiholin Hyb5 to lyse the cell. The corn genes are codon optimized for Bacillus subtilis, and the RBS ("SpoVG" BBa_K143021) has shown high strength in Bacillus subtilis. mRFP is used as a reporter because it is easily observed, easily measured, and expressed in Bacillus subtilis.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2436
    Illegal PstI site found at 1120
    Illegal PstI site found at 1606
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2436
    Illegal NheI site found at 1566
    Illegal PstI site found at 1120
    Illegal PstI site found at 1606
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2436
    Illegal BglII site found at 1056
    Illegal XhoI site found at 1940
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2436
    Illegal PstI site found at 1120
    Illegal PstI site found at 1606
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2436
    Illegal PstI site found at 1120
    Illegal PstI site found at 1606
    Illegal NgoMIV site found at 457
    Illegal NgoMIV site found at 944
    Illegal NgoMIV site found at 1739
    Illegal AgeI site found at 3667
    Illegal AgeI site found at 3779
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1943


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