Part:BBa_K5302011

pBBR-OmpA-mCherry

This work is derived from pBBR1MCS-2, pUC19-OmpA and pET-21b(+)-mCherry, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) 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.

Figure 1. Colony PCR results of pBBR1MCS-OmpA-mCherry

Figure 2. structure of OmpA, from genebank

Figure 3. SDS-PAGE analysis of pBBR1MCS-OmpA-mCherry expression in Escherichia coli Nissle 1917

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

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  Illegal EcoRI site found at 3884
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  Illegal EcoRI site found at 3884
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• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 3884
  Illegal EcoRI site found at 3932
  Illegal EcoRI site found at 4205
  Illegal EcoRI site found at 4608
  Illegal XbaI site found at 3189
  Illegal PstI site found at 2016
  Illegal PstI site found at 3177
  Illegal PstI site found at 4193
  Illegal NgoMIV site found at 2467
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  Illegal AgeI site found at 4411
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• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1379
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  Illegal SapI.rc site found at 2526