Part:BBa_K2812007
Coding sequence for Pyocin S5 with HlyA and His6-tag regulated by pBAD-ara promoter
Coding sequence for Pyocin S5, which is a bacteriocin produced by a specific strain of P. aeruginosa and is effective against other strains of P. aeruginosa. In this biobrick, Pyocin S5 is under control of the pBAD (araPb) promoter (BBa_K2812007). The sequence encodes for an effector component that consists of a receptor binding domain, a translocation domain and a killing domain. Furthermore it contains an immunity component which gives immunity against the bacteriocin. The HlyA tag is a secretion signal peptide which is used for secretion via the Type 1 secretion pathway of gram-negative bacteria. In order to facilitate protein purification, an N-terminal HIS-tag is present.
Usage and Biology
The working mechanism of Pyocin S5 involves making the target cell membrane permeable, causing leakage of intracellular materials. It has been demonstrated that is has antimicrobial activity against seven P. aeruginosa strains (DWW3, In3, In4, In7, In8, InA, and InB). P. aeruginosa PAO1, E. coli (BL21 (DE3), W3110, and ATCC15597) and S. aureus ATCC6538 were insensitive to pyocin S5. We as iGem Eindhoven 2018 aimed to use this biobrick for prevention of wound infections.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 208
Illegal BglII site found at 496
Illegal BglII site found at 2181 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1271
//cds/enzyme
//cds/enzyme/lysis
//cds/membrane
//cds/membrane/lysis
//chassis/prokaryote/ecoli
//collections/biofilm
//plasmid/expression
//plasmidbackbone/expression/inducible
//plasmidbackbone/proteinfusion
//plasmidbackbone/synthesis
//proteindomain
//proteindomain/cleavage
//proteindomain/degradation
//proteindomain/linker
//proteindomain/localization
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