Part:BBa_K5302014
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. intimin is an important outer membrane protein that includes several different domains: a lysozyme domain located in the periplasm, a β barrel domain embedded in the membrane, and an extracellular region at the C terminal or a "passenger" domain. This "passenger" domain is linked to the β barrel domain by a short linker sequence. intimin is classified as a reverse autotransporter, a subclass of the type 5 secretion system that transports the extracellular region or "passenger" of the C terminal through a compartment of the N terminal β barrel located in the outer membrane. The "passenger" domain of intimin forms a rod-like extension, consisting of four tandem bacterial immunoglobulin-like domains (D00-D0-D1-D2) with a C-terminal C-like domain (D3). The extension and rigidity of the D00-D0 domain of Intimin increases the possible radius of interaction with the protein, which improves intimin presentation after secretion to the bacterial surface.
Sequence and Features
- 10INCOMPATIBLE 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 - 12INCOMPATIBLE 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 - 21INCOMPATIBLE 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 - 23INCOMPATIBLE 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 - 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1379
Illegal SapI.rc site found at 2316
Illegal SapI.rc site found at 2526
biology | Escherichia coli Nissle 1917 |