This is a composite part consisting of both natural and synthetic DNA components
The pCASP circuit is a protein expression and secretion circuit. The system is currently only functional when expressed in Salmonella under SPI-1 expressing conditions. When SPI-1 is expressed the pCASP circuit will be activated for expression at the same time natural effectors would normally turn on and secrete. This circuit will express a protein of interest fused to the SptP167 secretion signal along with co-expression of the sicP chaperone. This combination is sufficient to direct the protein of interest to the export machinery and elicit secretion to the extracellular environment. In the initial configuration of this circuit the DH guanine exchange factor is used as a de-bugging secreted protein. Note: Some evidence exists of impassible substrates for the secretion machinery. Most of these proteins are highly thermodynamically stable domains that cannot be unfolded to transit through the secretion system.
This device was not designed with biobrick standard assembly in mind and has several of the scar sequence sites throughout the circuit.
The sicA promoter is a regulatory element in the Salmonella SPI-1 secretion system. It controls the expression of secreted effector proteins in the natural system and turns on late. Please reference Temme et al, 2008, JMB for exact dynamics of this promoter and associated feedback mechanisms. This promoter only functions in Salmonella strains containing SPI-1.
This part can only be used in salmonella strains.
Human Genomic DNA library clone with 2 point mutations
This protein coding frame is a Guanine Exchange Factor (GEF) domain from human eukaryotic with point mutations to make it constitutively active. This protein can be secreted at high titer via the Salmonella T3S using the Voigt Lab secretion circuit.
This part contains 2 PstI sites and is incompatible with biobrick standard assembly
Synthetic DNA from Oligonucleotide synthesis
A 3 repeat FLAG (DYKDDDDK) antigen epitope tag used for marking proteins of interest. Proteins carrying this tag can be easily detected using commercial western blotting reagents.
This sequence was designed as part of an oligonucleotide primer on each strand then annealed and ligated into the vector.
Synthetically created from oligonucleotide synthesis
The Tobacco Etch Virus (TEV) protease cleavage recognition sequence. This DNA sequence encodes an amino acid sequence recognized by TEV protease (ENLYFQS) for efficient cleavage. The sequence is flanked by glycines on the ends to increase flexibility and accessibility to the protease. After proteolytic processing the C-terminal fragment retains only an S residue, however by the design of this part it will leave an SG dipeptide which becomes the new N-terminus of the mature polypeptide.
This part was codon optimized for expression in E.Coli.
SicP-SptP chaperone-secretion tag pair from Salmonella type III secretion system
This part is the natural implementation of the SicP chaperone (BBa_J64002) and SptP secretion signal (BBa_J64003) from the Salmonella Type III Secretion System (T3S, the Salmonella T3S coding region is referred to as Salmonella Pathogeneity Island 1 [SPI-1]). In the natural implementation the coding regions of SicP and SptP overlap out of frame. This part captures that configuration for use in creating secreted protein / chaperone pairs.
This part is not compatible with biobrick standard assembly.
This is the natural RBS from the sicA chaperone (BBa_J6021) in the Salmonella Type III Secretion System.
This part was not designed with standard biobrick assembly in mind.
This is the T1 terminator from the pPROTet 6xHN vector commercially available from Clontech.
This sequence is not designed with biobrick standard assembly compatibility in mind.
Conversion of the iGEM parts registry to SBOL2.1