This is an HA tag without a start, but with 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 K112507 Wobble PCR of cc001/cc011 ( 54 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910,L) Product is pBca1256-K112507 -------------------------------------------- cc001 Forward Biobricking of {<HA>} CgATAgaattcATGagatcttacccatacgacgtcccagac cc011 Reverse biobricking of {<HA!} ccagtggatccttacccagcgtagtctgggacgtcgtatgggtaag
This BioBrick contains the coding sequence for silk tail with a Strep-tag behind it and ends with a stop codon. The Tail ??????. The Strep-tag, fused at the C terminal, is used both as a binding tag to use to inidicate its production and as a binding component to biotin.
The DNA sequence was codon-optimized for Bacillus Subtilis. Moreover the nucleotide sequence was optimized in order to accomplish the production standard of IDT. A couple of optimized codons were affected by this.
The amino acid sequence of the tail was obtained from MaSp2. The DNA sequence was synthesized by Integrated DNA Technologies (IDT).
hemagglutinin (HA) tag tail domain (YPYDVPDYA)
This is a HA tag tail domain designed to be at the C-terminus of 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 BioBrick contains the coding sequence for silk tail and ends with a stop codon. The Tail ....
This is His-tag with stop codon and RGD seguence. His-tag is used for detection of His-tagged proteins by anti-His antibodies. RGD is arginine-glycine-aspartic amino acid sequence, present on several extracellular matrix proteins. It is required for binding to integrins.
Part was constructed by primer annealing.
DNA ligase from Escherichia coli (LigA) with His-tag
This is an AP tag with stop codon, but no start. ---- 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 {<AP!} basic part K112601 Wobble PCR SC001/SC003 (69 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2472+ 910) Product is pBca1256-K112601 -------------------------------------------------- SC001 Forward Biobricking of <AP-tag! cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag SC003 Reverse Biobricking of <AP-tag! ccagtGGATCCttattcatgccattcaattttctgcgcttcaaaaatatcg
GFP - LasR protein complex with flag tags
This part contains a green fluorescent protein (GFP) as a non-reactive chemical compound (anchor) in order to neutralize the active reactive fixed to it, an inducer regulatory protein (LasR is responsible to induce pLas promoter in the presence of 6HSLO3 which is a chemical compound sythesized by an enzyme that LasI gene codes.) and a TEV protease cleavage site between them to assemble these two proteins. BBa_772001 and BBa_K772002 are assembled to be used as the trigger of detection in the whole system which ends with the intracellular transfection (or activation) of TEV protease. After the activation, it is planned that TEV protease will cleave this anchor-inducer protein complex to release free the reporter. In conclusion, in this system, the inducer would activate a transcription pathway on the targeted inducible gene or the reporter would indicate that there is a change in the whole system. This part contains flag tags extra from BBa_K772006 in the GFP region; because it is planned to be used in the Co-IP/SDS-PAGE assay to be isolated.
Note that there is an RBS sequence just before the protein sequence in order to eliminate the need of addition of it.
Cleavage site, LasR and GFP sequences are gathered from Registry.
VSV-G epitope tag tail domain (YTDIEMNRLGK)
This is a VSV-G epitope tag tail domain designed to be at the C-terminus of a protein. VSV-G tags are a type of epitope tag that can be used to measure the quantity of proteins.
A double TAA TAA stop codon was used. Rare codons from model organisms were avoided where possible.
This is a c-Myc tag head domain designed to be at the C-terminus of 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 tail 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.
Stable and soluble Histidine rich peptide. This part has a '''HindIII''' restriction site as extra.
PolyHb was desing taking into acount the following consideration: # avoided repeating the same residue in tandem to minimize local tRNA depleation # avoided Trp in tandem because of therir low solubility # we included Gly to avoid the formation of stable structures # included acidic and basic amino acids to increase solubility Sequence obtained by retro-translation. Codon and mRNA-secondary structure optimized for yeast.
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.
Synthetic DNA from Oligonucleotide synthesis
Stable and soluble Histidine rich peptide.
Retro-translated and optimized (codons and mRNA seconday structure) for yeast.
This is a S-tag tail domain designed to be at the C-terminus of a protein. S-tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
This is a FLAG tag with a stop codon, but no start 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 K112603 Wobble PCR SC004/SC006 (48 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2472+ 910) Product is pBca1256-K112603 ----------------------------------------------------- SC004 Forward Biobricking of <FLAG-tag! cgataGAATTCatgAGATCTgactacaaggatgacgacg SC006 Reverse Biobricking of <FLAG-tag! ccagtGGATCCttacttgtcgtcgtcatccttgtagtc
This is His-tag with stop codon. His-tag is used for detection of His-tagged proteins by anti-His antibodies. Cloned into pSB1AK3 vector.
This part is a C-terminal 6xHis-tag with a double stop-codon. It can be fused to the C-terminus of proteins to enable purification of fully-translated proteins.
In contrast to other 6x-His tag parts, this part uses the standard double stop codon TAA-TAA.
The part was designed using the histidin codon CAC and the double stop codons TAA-TAA.
V5 epitope tag tail domain (GKPIPNPLLGLDST)
This is a V5 epitope tag tail domain designed to be at the C-terminus of a protein. V5 tags are a type of epitope tag that can be used to measure the quantity of proteins.
A double TAATAA stop codon was included. Rare codons from common model organisms were avoided.
GFP - LacZ protein complex with flag tags
This part contains a green fluorescent protein (GFP) as a non-reactive chemical compound (anchor) in order to neutralize the active reactive fixed to it, a reporter device (LacZ enzyme catalyses the reaction of the cleavage of X-Gal which results with blue color change) and a TEV protease cleavage site between them to assemble these two proteins. BBa_772001 and BBa_K772002 are assembled to be used as the trigger of detection in the whole system which ends with the intracellular transfection (or activation) of TEV protease. After the activation, it is planned that TEV protease will cleave this anchor-inducer protein complex to release free the reporter. In conclusion, in this system, the inducer would activate a transcription pathway on the targeted inducible gene or the reporter would indicate that there is a change in the whole system. This part contains flag tags extra from BBa_K772006 in the GFP region; because it is planned to be used in the Co-IP/SDS-PAGE assay to be isolated.
Cleavage site, LacZ and GFP sequences are gathered from Registry.
This is a 6x His tag tail domain designed to encode the C-terminus of a protein. His tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
Used both His codons in order to avoid repeats. Also included a double TAATAA stop codon.
This is a myc tag without a start, but with 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 K112503 Wobble PCR of cc004/cc006 ( 54 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI,2227+ 910,L) Product is pBca1256-K112503 -------------------------------------------- cc004 Forward Biobricking of {<myc>} CgATAgaattcATGagatctGAACAAAAACTCATCTCAGAAGAGG cc006 Reverse Biobricking of {<myc!} ccagtggatccTTACAGATCCTCTTCTGAGATGAGTTTTTGTTC
This is a Strep tag tail domain designed to be at the C-terminus of a protein. Strep tags are a type of affinity tag that can be used to purify and/or measure the quantity of proteins.
This domain encodes the Strep tag sequence AWRHPQFGG. 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.
Stable and soluble '''Histidine''' rich peptide (designed in silico) This part has a '''HindIII''' restriction site as extra.
PolyWb was desing taking into acount the following consideration: # avoided repeating the same residue in tandem to minimize local tRNA depleation # avoided Trp in tandem because of therir low solubility # we included Gly to avoid the formation of stable structures # included acidic and basic amino acids to increase solubility Sequence obtained by retro-translation. Codon and mRNA-secondary structure optimized for yeast.
DNA ligase from Escherichia coli (LigA) with PT7, RBS and His-tag
'''TrpZipper''' is a small peptide that folds into a beta-hairpin secondary structure. The indole rings of the Trp form a hydrophobic core. The protein is water soluble and monomer.
Sequence obtained by retro-translation. Codon and mRNA-secondary structure optimized for yeast.
Tryptophan zippers: Stable, monomeric beta-hairpins [Cochran 2001]