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.
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.
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.
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.
Assembly scar between RFC[25] Biobrick and RFC[25] Biobrick.
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.
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.
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.
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.
Conversion of the iGEM parts registry to SBOL2.1