tthl (Laccase from Thermus thermophilus) coding sequence
UmuD derived N-terminal degradation tag; experimentation demonstrated that residues 1-29 of UmuD are sufficient to impart Lon recognition and degradation.
Derived from the 19 N-terminal amino acids of the UmuD protein. http://www.ncbi.nlm.nih.gov/sites/entrez?Db=gene&Cmd=ShowDetailView&TermToSearch=945746&ordinalpos=1&itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum
hsl1 - cell cycle dependent degradation tag in yeast
This BioBrick is a fragement of the yeast hsl1 protein. Fused to an artificial transcription factor it tiggers the degradation in??? phase via the ???.
To enable the fusion with the protein in frame the vector of this BioBrick has no base pair in between the restriction sites and the BioBrick. Furthermore, this coding sequence does not include a start codon. For more information about this issus, see: Phillips, I.E. and Silver, P.A. "A new Biobrick Assembly Strategy Designed for Facile Protein Engineering." DSpace http://hdl.handle.net/1721.1/32535 (2006).
Fusion protein of S-Layer SbpA with T7/lac and BisdA and BisdB
ecol laccase from E. coli with T7 promoter, RBS and His-tag
E.coli laccase ORF with T7, RBS and HIS tag
Fusion protein of SbpA, NADP+ Oxidoreductase, BisdA and BisdB with PT7/lac
PT7 + RBS + SbpA || Red || BisdA || BisdB
AANDENYNYADAS. (Fast) SsrA degradation tag.
This sequence codes for the amino acid sequence AANDENYNYADAS, a variation of the WT SsrA tag sequence, which, when fused to the C-terminal of proteins, will make the protein susceptible to fast degradation through SspB-mediated binding to the ClpX protease. The following rates of degradation of this tag are pulled from the corresponding references below: ~2 Vmax/ [Clpx6] min-1 from (1). See the following references for further information on degradation rates and mechanisms of this tag: (1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. (2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240.
C-terminal tag. Degradation rate is fast. Variations of this sequence yield different degradation rates(see Parts BBa_M0050, BBa_M0052, BBa_M0053). Sequence derived from reverse translation of AANDENYNYADAS sequence using codon usage optimized for E. coli. Three C-terminal aa's (DAS in this case) are necessary and sufficient for ClpX binding and degradation. Upstream aa sequence serves as a binding site for SspB, which guides rapid binding to ClpX. See sources on the main part page for further information about the mechanism of the system. NOTE: this sequence has only the amino acid sequence for the tag and bears NO STOP CODONS, so be sure to inlclude them if you use this sequence.
This variation of the SsrA tag was studied by McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701.
bhal laccase from Bacillus halodurans with T7 promoter, RBS and His-tag
bhal laccase from Bacillus halodurans with T7 and HIS
tthl laccase from Thermus thermophilus with T7 promoter, RBS and His-tag
tthl (Laccase from Thermus thermophilus) with T7, RBS and HIS tag
AANDENYADAS (moderately fast) SsrA degradation tag.
This sequence codes for the amino acid sequence AANDENYADAS, a slower variation of the WT SsrA tag sequence, which, when fused to the C-terminal of proteins, will make the protein susceptible to moderately fast degradation through SspB-mediated binding to the ClpX protease. The following rates of degradation of this tag are pulled from the corresponding references below: ~1 Vmax/ [Clpx6] min-1 from (1). See the following references for further information on degradation rates and mechanisms of this tag: (1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. (2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240.
C-terminal tag. Degradation rate is moderately fast. Variations of this sequence yield different degradation rates(see Parts BBa_M0050, BBa_M0051, BBa_M0053). Sequence derived from reverse translation of AANDENYADAS sequence using codon usage optimized for E. coli. Three C-terminal aa's (DAS in this case) are necessary and sufficient for ClpX binding and degradation. Upstream aa sequence serves as a binding site for SspB, which guides rapid binding to ClpX. See sources on the main part page for further information about the mechanism of the system. NOTE: this sequence has only the amino acid sequence for the tag and bears NO STOP CODONS, so be sure to include them if you use this sequence.
This variation of the SsrA tag was studied by McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. SsrA tags are prevalent in E. coli.
cin8 - cell cycle specific degradation tag in yeast
This BioBrick is the xxx domain of the yeast cin8 protein. It has been shown that this region of part of the protein is suffient to get xxx specific degradation via the APC complex.
To enable the fusion with other domains in frame the vector of this BioBrick has no base pair in between the restriction side and the BioBrick. Furthermore, this coding sequence does not include a start codon.<br\n> For more information about this issus, see:<br\n> Phillips, I.E. and Silver, P.A. "A new Biobrick Assembly Strategy Designed for Facile Protein Engineering." <br\n> DSpace http://hdl.handle.net/1721.1/32535 (2006).
bpul (laccase from Bacillus pumilus) with T7 promoter, RBS and HIS tag
bpul (Laccase from Bacillus pumilus) with T7, RBS and HIS tag
The MerB gene codes for the proetin which function is to cleave the organic part of the molecule of methyl-mercury
AANDENYALAA. (Very fast) SsrA degradation tag.
This sequence codes for the amino acid sequence AANDENYALAA, which when fused to the C-terminal of proteins, will make the protein susceptible to very fast degradation through SspB-mediated binding to the ClpX protease. The following rates of degradation of this tag are pulled from the corresponding references below: ~5 Vmax/ [Clpx6] min-1 from (1) ~0.5%/min on log scale from (2) ~1 min half life from (3) See the following references for further information on degradation rates and mechanisms of this tag: (1)McGinness, Baker, Sauer. 2006. Mol. Cell. 22:701. (2)Flynn et al 2003. Mol. Cell. 11: 671. Flynn et al. 2001. PNAS 98(19): 10584. Anderson et al 1998. App. Env. Microbiol. 64(6):2240.
C-terminal tag. Degradation rate is very fast. Deviations from this sequence in key amino acids will lower degradation rates (see Parts BBa_M0051, BBa_M0052, BBa_M0053). Three C-terminal aa's (LAA in this case) are necessary and sufficient for ClpX binding and degradation. Upstream aa sequence serves as a binding site for SspB, which guides rapid binding to ClpX.
C-terminal degradation tags are commonly found in high turnover proteins in Escherichia coli.
Fusion protein of S-Layer SgsE with T7/lac and BisdA and BisdB
cln2 PEST destabilization domain for rapid protein turnover
This BioBrick is the xxx domain of the yeast cln2 protein. It has been shown that this region of part of the protein is suffient to get xxx specific degradation via the APC complex. <br\n>
Fusion protein of SgsE, NADP+ Oxidoreductase, BisdA and BisdB with PT7/lac
PT7 + RBS + SgsE || Red || BisdA || BisdB ; RCU 25
Appl Environ Microbiol, 1998 June; 64(6):2240-2246
tthl laccase from T. thermophilus with constitutive promoter J23100, RBS and His-tag
tthl laccase (T. thermophilus) with constitutive promoter and HIS tag
xccl laccase from Xanthomonas campestris with T7 promoter, RBS and His-tag
xccl laccase from Xanthomonas campestris with pT7 and HIS tag
Xanthomonas campestris pv. campestris B100