iGEM Team Aachen BioBricks
RFC [25] Version of E0020
This part is an RFC [25]-compatible version of BBa_E0030. The start and stop codons have been removed to make it RFC [25]-compatible and the part is flanked by the RFC [25] prefix- and suffix-sequences.
The coding sequence encodes EYFP (enhanced yellow fluorescent protein) which is derived from A. victoria GFP. The excitation is 512 nm and the emission is 534 nm.
RFC [25] - compatible dark quencher based on K1319000 (E0030)
This part is an RFC [25] dark quencher that is based upon K1319000 (the RFC [25] version of E0030/EYFP).
Two mutations were introduced that eliminated fluorescence:
RFC [25] -compatible dark quencher based on K1319000 (E0030)
This part is an RFC [25] dark quencher that is based upon K1319000 (the RFC [25] version of E0030/EYFP).
Three mutations were introduced that eliminated fluorescence:
human galectin-3, codon optimized for E. coli
The trunkated galectin-3 is a 26 kDa protein that binds certain LPS patterns.
TEV protease with anti-self cleavage mutation S219V, codon optimized for E. coli
This part is a TEV protease in RFC25 that was codon-optimized for expression in E. coli. The part contains the S219V anti-self cleavage mutation.
The TEV Protease (also known as Tobaco Edge Virus nuclear inclusion a endopeptidase) is a highly sequence specific cysteine protease from the Tobacco Edge Virus (TEV). The protease is highly sequence specific. The consensus sequence for the cut is ENLYFQ\S with \ denoting the cleaved peptide bond. This sequence can be found in the part K1319016.
ENLYFQ\S is the optimal cleavage site with the highest activity but the protease is also active to a greater or lesser extent on a range of substrates. The highest cleavage is of sequences closest to the consensus EXLYΦQ\φ where X is any residue, Φ is any large or medium hydrophobic amino acid and φ is any small hydrophobic amino acid.
The TEV protease is commonly used as a biochemical tool to cleave affinity tags from purified proteins like His-Tags. The high specifity makes the protease relatively non-toxic in vitro and in vivo. The molecular weight of the TEV protease is 27 kDa.
IPTG-induced and T7-driven expression of TEV protease
This protein generator produces TEV protease when induced with IPTG in a DE3 strain or if combined with a T7 RNA-Polymerase generator.
Constitutive expression of K1319000
This part expresses K1319000 behind a J23101 constitutive promotor.
Constitutive expression of K1319001
This part expresses K1319001 behind a J23101 constitutive promotor.
Constitutive expression of K1319002
This part expresses K1319002 behind a J23101 constitutive promotor.
Constitutive expression of EGFP-REACh1 fusion protein
This part expresses a E0040.K1319001 fusion protein (EGFP-REACh1) behind a J23101 promotor.
Constitutive expression of EGFP-REACh2 fusion protein
This part expresses a E0040.K1319002 fusion protein (EGFP-REACh1) behind a J23101 promotor.
Constitutive expression of EGFP-EYFP fusion protein
This part expresses a E0040.K1319000 fusion protein (EGFP-EYFP) behind a J23101 promotor.
TEV protease cleavage site
This sequence codes for a TEV protease cleavage site.
ENLYFQ\S is the optimal cleavage site with the highest activity but the protease is also active to a greater or lesser extent on a range of substrates. The highest cleavage is of sequences closest to the consensus EXLYΦQ\φ where X is any residue, Φ is any large or medium hydrophobic amino acid and φ is any small hydrophobic amino acid.
LasI induced iLOV
This device produces iLOV (K660004) in response to a quorum sensing input.
Translational unit of mRFP-galectin3-his
This part is a translational unit of a mRFP-galectin-3-his (B0032.E1010.K1319003.K1319016.B0015).
IPTG inducible iLOV
This part can be used for IPTG-induced expression of K660004 (iLOV).
- Part Name
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- Sequence
- Short Description (60 characters on what the DNA does)
- Long Description (Longer description of what the DNA does)
- Design considerations
<groupparts>iGEM14 Aachen</groupparts>
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