Part:BBa_K311004
Complement of Ethanolamine Utilization Microcompartment proteins from Salmonella LT2
A total of 5 micro-compartment genes from the Salmonella LT2 operon are stacked and cloned into the pSB1C3 plasmid. They are arranged as two sets of two genes - EutMN and EutLK - in a single operon with one gene EutS) which is upstream to the pairs. EutS, EutMN and EutLK are each flanked directly upstream by a mutated Lac promoter which is constitutively active. The Eut genes were amplified by PCR from LT2 genomic DNA using primers that would introduce restriction sites (5' Bgl II and 3' Not I) flanking the genes.
The fluorescence microscopy images above show E. coli cells transformed with a plasmid containing EutSMNLK and a plasmid containing GFP. The E. coli in the two images on the left were transformed with a chimeric GFP that has a predicted Eut Shell targeting sequence at the N-terminus. By introducing this tag, the GFP is believed to be localized within the microcompartment. These areas are indicated by yellow arrows. The DIC images are a control to visualize the boundaries of the cells such as the image with the EutSMNLK and the chimeric GFP.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 476
Illegal NotI site found at 1386 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 134
Illegal BglII site found at 689
Illegal BglII site found at 1599
Illegal XhoI site found at 484
Illegal XhoI site found at 1394 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1313
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 901
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