The S???Tag sequence is a novel fusion peptide tag for recombinant proteins that allows detection by a rapid, sensitive homogeneous assay or by colorimetric detection in Western blots. Proteins can also be rapidly purified from crude extracts using a one step affinity separation method. The system is based on the strong interaction between the 15aa S???Tag and 103aa S-protein.
Glutathione S-transferase internal domain
This is a GST internal domain designed to be between protein domains. GST domains are a type of affinity domain that can be used to purify and/or measure the quantity of proteins.
Rare codons in common model organisms were avoided along with restriction sites used in various assembly standards.
c-Myc special internal domain (EQKLISEEDL)
This is a c-Myc tag special internal domain designed to be within a protein. c-Myc tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
This domain encodes the c-Myc sequence EQKLISEEDL. The codons at each amino acid position were selected to avoid rare codons in organisms like E. coli, humans, mouse, B. subtilis, C. elegans, Streptomyces coelicolor and other common model organisms and/or industrial hosts.
VSV-G epitope tag special internal domain (YTDIEMNRLGK)
This is a VSV-G epitope tag special internal domain designed to be used between protein domains. VSV-G tags are a type of epitope tag that can be used to measure the quantity of proteins.
Rare codons from model organisms were avoided where possible.
hemagglutinin (HA) tag special internal domain (YPYDVPDYA)
This is a HA tag special internal domain designed to be within a protein. HA tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
This domain encodes the HA sequence YPYDVPDYA. The codons at each amino acid position were selected to avoid rare codons as much as possible in organisms like ''E. coli'', humans, mouse, ''B. subtilis'', C. elegans, ''Streptomyces coelicolor'' and other common model organisms and/or industrial hosts. Another factor in codon selection was avoiding avoid repeat sequences.
This is a 6x His tag special internal domain is designed to be between protein domains. His tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
Used both Histidine codons to minimize repeats.
FLAG octapeptide special internal domain (DYKDDDDK)
This is a FLAG tag octapeptide special internal domain designed to be internal to the sequence of a protein. FLAG tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
The FLAG sequence AspTyrLysAspAspAspAspLys (DYKDDDDK) is one of the most commonly used FLAG tag sequences. Each of the amino acids in this octapeptide have two possible codon choices. For each amino acid, neither of the two codon choices appear to be rare in the common model organisms. So codons were selected to minimize repeats.
This is an HA tag without a start or a stop codon. It is in BBb format. ---- This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
Construction of {<HA>} basic part K112506 Wobble PCR of cc001/cc010 ( 51 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910,L) Product is pBca1256-K112506 -------------------------------------------- cc001 Forward Biobricking of {<HA>} CgATAgaattcATGagatcttacccatacgacgtcccagac cc010 Reverse biobricking of {<HA>} ccagtggatcccccagcgtagtctgggacgtcgtatggg
This is FLAG tag with no start and no stop codon ---- This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
Construction of {<FLAG>} basic part K112602 Wobble PCR SC004/SC005 (45 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2472+ 910) Product is pBca1256-K112602 --------------------------------------------------- SC004 Forward Biobricking of <FLAG-tag> cgataGAATTCatgAGATCTgactacaaggatgacgacg SC005 Reverse Biobricking of <FLAG-tag> ccagtGGATCCcttgtcgtcgtcatccttgtagtc
Import enhancer - Polyarginine (trojan peptide)
Trojan peptides are short sequences that penetrate through the plasma membrane inside the cell without the need of any receptor or endocitosis process [Derossi 1998]. They can be used to increase the efficiency with which a protein enters a cell. This part is Polyarginine, and has to be used just before the coding sequence of the peptide that you want to be import enhanced. (refer to ''Characterisation of cell-penetrating peptide-mediated peptide delivery'' [Jones et al 2005] for more information about polyarginine)
Sequence obtained by retro-translation. Codon and mRNA-secondary structure optimized for yeast.
Characterisation of cell-penetrating peptide-mediated peptide delivery [Jones et al 2005]
This is the polypeptide protein FLAG tag. It is used in antibody binding assays if there is no antibody against the studied protein. It should be put at either the N terminus or the C terminus of the protein This part uses the modified Silver BioBrick prefix and suffix to allow for protein construction.
Tags are almost always used in conjunction with another protein (the one being studied), so we used the modified Silver BioBrick prefix and suffix to make the construction of funcitonal fusion peptides possible
This sequence is easily recognized by many antibodies.
Strep-tag II special internal domain (WSHPQFEK)
This is a Strep tag tail domain designed to be internal to a protein. Strep tags are typically used for detection, purification and immobilization of proteins.
This domain encodes the Strep tag sequence WRHPQFGG. The codons at each amino acid position were selected to avoid rare codons as much as possible in organisms like ''E. coli'', humans, mouse, ''B. subtilis'', ''C. elegans'', ''Streptomyces coelicolor'' and other common model organisms and/or industrial hosts. Another factor in codon selection was avoiding avoid repeat sequences.
V5 epitope tag special internal domain (GKPIPNPLLGLDST)
This is a V5-tag special internal domain designed to between protein domains. V5 tags are a type of epitope tag that can be used to measure the quantity of proteins.
Rare codons from common model organisms were avoided.
This is the polypeptide protein HA tag. It is used in antibody binding assays if there is no antibody against the studied protein. It should be put at either the N terminus or the C terminus of the protein This part uses the modified Silver BioBrick prefix and suffix to allow for protein construction.
This is a myc tag without a stop or a stop codon. It is in BBb format. ---- This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
Construction of {<myc>} basic part K112502 Wobble PCR of cc004/cc005 ( 51 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910,L) Product is pBca1256-K112502 -------------------------------------------- cc004 Forward Biobricking of {<myc>} CgATAgaattcATGagatctGAACAAAAACTCATCTCAGAAGAGG cc005 Reverse Biobricking of {<myc>} ccagtggatccCAGATCCTCTTCTGAGATGAGTTTTTG
This codes for six Histidine residues to allow for His column protein purification. It will work best if put at either the N or C terminus of your protein construct. This part uses the modified Silver BioBrick prefix and suffix to allow for protein construction.
This part uses the modified Silver BioBrick prefix and suffix to allow for protein construction.
This is a chitin binding domain (CBD) designed to be at between protein domains. Chitin binding domains are a type of affinity tag that can be used to purify proteins.
Rare codons from common model organisms as well as restriction sites used in various assembly standards were removed.
Import enhancer - HIV TAT penetratin (trojan peptide)
Trojan peptides are short sequences that penetrate through the plasma membrane inside the cell without the need of any receptor or endocitosis process [Derossi 1998]. They can be used to increase the efficiency with which a protein enters a cell. This part is the penetratin from the HIV TAT protein, and has to be used just before the coding sequence of the peptide that you want to be ''import enhanced''.
S-tag special internal domain (KETAAAKFERQHMDS)
This is a S-tag special internal domain designed to between protein domains. S-tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
Rare codons from various model organisms were avoided where possible.