Plasmid

Part:BBa_K5302014

Designed by: Bojun Yang   Group: iGEM24_USTC   (2024-10-01)
Revision as of 09:36, 1 October 2024 by Xiaotian (Talk | contribs)


pBBR-intimin-mCherry

This work is derived from pBBR1MCS-2, pUC19-intimin-spytag and pET-21b(+)-mCherry, and it has undergone codon optimization. This composite part combines intimin-spytag(approximately 49.5kda) and mCherry(26.7kda), and we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express mCherry as a fluorescence labeling.The plasmid uses lac promotor and has kanamycin resistence.

Jamboree Program
Figure 1. structure of intimin, from genebank

Jamboree Program
Figure 2. Colony PCR results of pBBR1MCS-intimin-mCherry

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 4483
    Illegal EcoRI site found at 4531
    Illegal EcoRI site found at 4804
    Illegal PstI site found at 2016
    Illegal PstI site found at 3494
    Illegal PstI site found at 3742
    Illegal PstI site found at 4189
    Illegal PstI site found at 4792
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4483
    Illegal EcoRI site found at 4531
    Illegal EcoRI site found at 4804
    Illegal PstI site found at 2016
    Illegal PstI site found at 3494
    Illegal PstI site found at 3742
    Illegal PstI site found at 4189
    Illegal PstI site found at 4792
    Illegal NotI site found at 1057
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4483
    Illegal EcoRI site found at 4531
    Illegal EcoRI site found at 4804
    Illegal BglII site found at 1803
    Illegal XhoI site found at 5149
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 4483
    Illegal EcoRI site found at 4531
    Illegal EcoRI site found at 4804
    Illegal PstI site found at 2016
    Illegal PstI site found at 3494
    Illegal PstI site found at 3742
    Illegal PstI site found at 4189
    Illegal PstI site found at 4792
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 4483
    Illegal EcoRI site found at 4531
    Illegal EcoRI site found at 4804
    Illegal PstI site found at 2016
    Illegal PstI site found at 3494
    Illegal PstI site found at 3742
    Illegal PstI site found at 4189
    Illegal PstI site found at 4792
    Illegal NgoMIV site found at 2467
    Illegal NgoMIV site found at 2750
    Illegal NgoMIV site found at 5740
    Illegal AgeI site found at 5010
    Illegal AgeI site found at 5580
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1379
    Illegal SapI.rc site found at 2316
    Illegal SapI.rc site found at 2526


[edit]
Categories
//plasmid
Parameters
biologyEscherichia coli Nissle 1917