sequence containing necessary DNA for controlled chemotaxis
DNA includes switching mechanisms and componenents necssary for directed motility towards aspartate or phenylalanine (does not include wild-type CheW, which may be inserted between restriction sites on opposite side of inversion sequence). Sequence must be placed in strain with all receptors knocked out (tar, tsr, tap, trg, ser) as well as CheW. Switching mechanism is irreversibly activated by arabinose. Strain switches from phenylalanine chemotaxis to aspartate chemotaxis.
CheW* (VM108), Tar receptor codon-optimized to avoid recombination.
CheW with VM108 mutation for orthogonal interaction with Tar EA400
CheW was cloned out of the genome and mutated on plasmid
Part contains CheW with valine to methionine mutation at position 108 to facilitate an orthogonal interaction with a Tar chemotaxis receptor with a EA400 mutation. CheW VM108 will have only limited interaction with wild-type Tar.
Inverter component includes araC+pBAD+FimE+T1/T2/T3/T4
Component to be used in nucleotide sequence inversion. Addition of arabinose activates switching mechanism. Switch is irreversible.
Invertible sequence of dna includes Ptrc promoter
Seuqence is inverted by the FimE protein. Inside recognition sequence is a Ptrc promoter sequence. Switch is irreversible.
ribosome binding site for transcription initiation
terminator sequence to end transcription
Ribosome binding site for transcription initiation
Promoter sequence for constituitive expression
Tar EA400 has orthogonal interaction with CheW VM108
Tar (aspartate-sensitive chemotaxis receptor) mutated (EA400) to have orthogonal interaction with CheW VM108. Has minimal interaction with wild-type CheW.
Mutation was originally evolved in Tsr (EA402), this residue occurs in position 400 after sequence alignment.
Tar mutated to respond to phenylalanine instead of aspartate
Tar (aspartate-sensitive chemotaxis receptor) with mutations to respond to phenylalanine. Protein has minimal response to aspartate.
Conversion of the iGEM parts registry to SBOL2.1