Red light triggered Kill Switch for plants translation unit (PhyB/PIF3 version)
Assembly scar between RFC[10] coding/non-coding Biobrick and RFC[25] N-part Biobrick (not RFC[25] standard prefix!). Formally it has the same scar like the assembly scar between RFC[10] coding/non-coding Biobrick and RFC[10] coding Biobrick.
Membrane-associated Thermonuclease NucA with TEV site and SV40 NLS in RFC[25] N-Part
This part encodes a membrane-associated Thermonuclease NucA for the chassis ''Physcomitrella patens''. NucA is a endo/exonuclease that degrades ssDNA/dsDNA/ssRNA/dsRNA. It cannot execute its deadly potential when it is bound to the cytoplasmatic membrane. The region between NucA and the transmembrane domain consists a long linker with a TEV cleavage site and a SV40 nuclear localization signal (NLS). By cleaving the TEV cleavage site with a TEV Protease the nuclease NucA translocates in the nucleus due to its SV40 NLS and degrades the genomic DNA resulting a inevitable induction of programmed cell-death (PCD). This construct is flanked by RFC[25] N-part prefix and suffix. Note: This means only protein fusions to the C-terminus of this part is possible, adding promoters (typically RFC[10]) into the prefix or terminators/IRES (typically RFC[10]) into the suffix of this part is nevertheless possible.
Eukaryotic light-switchable TEV Protease translational unit (PhyB/PIF3 based) with downstream IRES
This part encodes a light-switchable TEV Protease system consisting of two fusion proteins ([http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159106 BBa_K1159106] and [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159107 BBa_K1159107]) seperated by an polioviral IRES [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159300 BBa_K1159300] which allows polycistronic expression in eukaryotic systems. You can easily express a third protein by simply clone your desired protein in the suffix of this part because this part ends with a polioviral IRES which allows cap-independent translation. After addition of your desired protein you can express this system by inserting your favored promoter/terminator in the prefix/suffix your construct. For further information, please see under the subparts linked in the text.
Assembly scar between RFC[25] Biobrick and RFC[25] Biobrick.
Polioviral Internal Ribosome Entry Site (IRES) for polycistronic expression in eukaryotes
This part contains the Internal Ribosome Entry Site (IRES) of the human poliovirus 1 Mahoney strain which allows polycistronic expression systems in eukaryotic systems. This part is flanked by RFC[10] pre- and suffix. By cloning this part betweens CDSs, many proteins/peptides can expressed by a single promoter and terminator.
Assembly scar between RFC[25] Biobrick and RFC[10] non-coding Biobrick.
Thermonuclease with N-terminal SV40 NLS and flexible linker with TEV site in RFC[25]
This part encodes a micrococcal nuclease (Thermonuclease) with N-terminal SV40 Nuclear Localization Signal (SV40 NLS) and a flexible linker with a TEV cleavage site. Thermonuclease degrades ssDNA/dsDNA/ssRNA/dsRNA. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
TMD with N-terminal Signal Peptide of SERK Receptor from ''P. patens'' in RFC[25] N-Part
This part codes for the Transmembrane Domain with N-terminal Signal Peptide of SERK Receptor from ''P. patens'' in RFC[25]. This construct is flanked by RFC[25] N-part prefix and suffix. Note: This means only protein fusions to the C-terminus of this part is possible, adding promoters (typically RFC[10]) into the prefix or terminators/IRES (typically RFC[10]) into the suffix of this part is nevertheless possible.
Eukaryotic light-switchable TEV Protease translational unit (PhyB/PIF3 based)
This part encodes a light-switchable TEV Protease system consisting of two fusion proteins ([http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159106 BBa_K1159106] and [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159107 BBa_K1159107]) seperated by an polioviral IRES [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159300 BBa_K1159300] which allows polycistronic expression in eukaryotic systems. To express this system you only have to insert your favored promoter/terminator in the prefix/suffix of this part. For further information, please see under the subparts linked in the text.
Signal Peptide of SERK Receptor from Physcomitrella patens in RFC[25]
This part codes for the signal peptide from the SERK receptor (Somatic Embryogenesis RECEPTOR KINAS) of ''Physocmitrella patens''. The signal peptide induces translocation of the protein into the Endoplasmatic Reticulum (ER) for secretion or membrane integration. Protein fused to the C-terminus of this part should be secreted or otherwise if the part fused region contains a transmembrane domain it will be integrated in cytoplasmatic membrane. This part starts with a consensus sequence for ''Physcomitrella patens'' which allows efficient translation start. Furthermore, this part is flanked by RFC[25] N-part pre- and suffix for fusions to the C-terminus of this part.
Transmembrane domain of SERK Receptor from Physcomitrella patens in RFC[25]
This part is encoding the transmembrane region of the SERK Receptor from ''Physcomitrella patens''. For membrane translocation a N-terminal signal peptide is still necessary for translocation through the ER membrane. This part is flanked by RFC[25] pre- and suffix.
Transmembrane Domain of SERK Receptor from ''P. patens''
34 AA flexible linker with C-terminal TEV cleavage site (GGGGS)x5-TEV-site-linker) in RFC[25]
This part encodes a 34 amino acids long flexible linker ((GlyGlyGlyGlySer)x5) with a C-terminal cleavage site for the TEV Protease. For further protein fusions this part is flanked by RFC[25} pre- and suffix.
Assembly scar between RFC[10] coding/non-coding Biobrick and RFC[25] Biobrick (not RFC[25] N-part!).
PhyB(2-908 AA)_36AALinker_C-TEV-Protease (First part for a light-switchable TEV-Protease) in RFC[25]
This parts encodes are fusion protein consisting the photosensitive core of PhyB (2-908 AA) at the N-terminus and the C-terminal half of TEV Protease at its C-terminus. Both parts are seperated by a 36 amino acids long flexible linker. This part is flanked by RFC[25] pre- and suffix to allow further protein fusions. Expressed with the second essential protein [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159315] or [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159315] containing the part which interacts with PhyB and the N-terminal half of the splitted TEV Protease, it form a light-switchable TEV Protease which is activated by red light and inactivated by far-red light. '''''Note: PhyB requires Phycocyanobilin or Phytochromobilin as a cofactor to fulfill its function as photoswitchable protein; you can reach it by adding exogenous Phycocyanobilin/Phytochromobilin in the medium or by inserting genes for the Phycocyanobilin/Phytochromobilin synthesis. See here for more information: [http://2012.igem.org/Team:TU_Munich/Project/Light_Switchable_Promoter#Biosynthesis_of_Phycocyanobilin] '''''
N-TEV-Protease_36AALinker_PIF3 (Second part for a light-switchable TEV-Protease) in RFC[25]
This parts encodes a fusion protein consisting the N-terminal half of the TEV Protease as its terminus and first 100 amino acids of PIF3 as its C-terminus. PIF3 interacts with PhyB in the presence of red light. This reaction is reversible by far-red light. Both parts are seperated by a 36 amino acids long flexible linker. This part is flanked by RFC[25] pre- and suffix to allow further protein fusions. By expressing this part together with the second essential protein [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159106 BBa_K1159106] that contains the part, which interacts with PIF3, and the C-terminal half of the splitted TEV Protease, they form together a light-switchable TEV Protease system which can be activated by red light and inactivated by far-red light.
Micrococcal nuclease (Thermonuclease) with N-terminal SV40 NLS in RFC[25]
This part encodes a micrococcal nuclease (Thermonuclease) with N-terminal SV40 Nuclear Localization Signal (SV40 NLS). Thermonuclease degrades ssDNA/dsDNA/ssRNA/dsRNA. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
C-terminal half of TEV Protease S219V mutant (for Split-TEV-Protease applications) in RFC[25]
C-terminal half of the TEV Protease (autolysis resistant S219V mutant) can be used for Split-TEV-Protease applications where autolysis of the reconstituted TEV Protease is not wanted. The necessary N-terminal half can be found under BBa_K1159101. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
Phytochrome B (PhyB, first 908 N-terminal amino acids)
improved version of BBa_K365002 to allow N- and C-terminal protein fusions using RFC 25
Essential Arg for photoconvertible binding of PIF3/6
Essential Gly for photoconvertible binding of PIF3/6
Essential Cys for chromophore attachement of phycocyanobilin/phytochromobilin
This part is a linker, it can be used to connect two parts and add additional space between these parts. That can be necessary to avoid interactions between these parts. The GSAT linker was create to connect our protein domains Fok_a and Fok_i to an function final construct.
Designed for the fusion of proteins or peptides via in frame cloning, according to RFC25.
N-terminal half of TEV Protease S219V mutant (for Split-TEV-Protease applications) in RFC[25]
N-terminal half of the TEV Protease (autolysis resistant S219V mutant) can be used for Split-TEV-Protease applications where autolysis of the reconstituted TEV Protease is not wanted. The necessary C-terminal half can be found under BBa_K1159102. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
N-terminal Phytochrome Interacting Factor 3 (PIF3) in RFC[25]
N-terminal Phytochrome Interacting Factor 3 (PIF3) is known to be recognized by Phytochrome B (PhyB) under red light exposition. This interaction between PhyB and PIF3 can be reverted by irradiation with far-red light. Only the first 100 amino acid residues of PIF3 is necessary for light-dependent binding to PhyB. N- and C-terminal protein fusions seems not to affect the binding of PhyB. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
Mature Nuclease NucA from Staphylococcus aureus (Thermonuclease) in RFC[25]
Thermonuclease is a endo-exonuclease from ''Staphylococcus aureus'' that degrades dsDNA, ssDNA, dsRNA and ssRNA. This part is coding the mature form of the nuclease, also called NucA. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
Polypeptide containing SV40 nuclear localization sequence (SV40 NLS) for nuclear translocation
designed by oligo hybridisation based on plasmid "pGADT7 AD" from Clontech, w/ RFC25-pre and suffix for protein fusions
SV40 nuclear localization sequence (recognized by importin alpha)
Conversion of the iGEM parts registry to SBOL2.1