Difference between revisions of "Part:BBa K5302011"

Revision as of 09:25, 1 October 2024

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]
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
12
INCOMPATIBLE WITH RFC[12]
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 PstI site found at 2016
Illegal PstI site found at 3177
Illegal PstI site found at 4193
Illegal NotI site found at 1057
21
INCOMPATIBLE WITH RFC[21]
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 BglII site found at 1803
Illegal BamHI site found at 3195
Illegal XhoI site found at 4550
23
INCOMPATIBLE WITH RFC[23]
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
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
Illegal NgoMIV site found at 2750
Illegal NgoMIV site found at 5266
Illegal AgeI site found at 4411
Illegal AgeI site found at 5106
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

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