Membrane-anchored SpyTag with secretory SpyCatcher_nLuc reporter fusion translational unit
This part encodes a translational unit consisting two fusion proteins, a Membrane-anchored SpyTag and a secretory SpyCatcher_nLuc (IgKappa-SigP_SpyCatcher_nLuc) reporter fusion to show if post-translational protein fusion like reaction happens between the membrane-localized tag protein and the secretory fusion protein containing N-terminal SpyCatcher and C-terminal NanoLuc Luciferase. As a translational unit both fusion proteins are seperated by a polioviral IRES to allow bicistronic 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.
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[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.
Assembly scar between RFC[25] Biobrick and RFC[10] non-coding Biobrick.
Membrane-anchored SpyTag in RFC[25] N-Part
This part codes for a membrane-anchored version of SpyTag [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159201 BBa_K1159201] for the chassis ''Physcomitrella patens''. Please see subparts for for further description. 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.
Secretory SpyCatcher_nLuc fusion (IgKappa-SigP_SpyCatcher_nLuc) in RFC[25] N-Part
Secretory Fusionprotein consisting N-terminal SpyCatcher and C-terminal nLuc (IgKappa-SigP_SpyCatcher_nLuc) in RFC[25] N-Part. 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.
Assembly scar between RFC[25] Biobrick and RFC[25] Biobrick.
SERK-Receptor TMD with C-terminal GFP and N-terminal Strep-tag II TEV-site linker in RFC[25]
This part codes for the Transmembrane Domain of SERK Receptor from ''Physcomitrella patens'' with a C-terminal (intracellular) GFPmut1 fusion and a N-terminal fusion to a linker containing ''Strep''-tag II and a TEV cleavage site; SERK TMD and GFPmut1 are seperated by a middle long linker. This part is flanked by RFC[25] pre- and suffix for further protein fusions. Note: A signal peptide is needed for membrane integration.
Secretory SpyCatcher (IgKappa-SigP_SpyCatcher) in RFC[25] N-Part
This part encodes the SpyCatcher [http://parts.igem.org/Part:BBa_K1159200 BBa_K1159200] with a N-terminal signal peptide for secretion (Ig Kappa signal peptide from ''Mus musculus'') which also works in moss ''Physcomitrella patens''. 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.
Secretory SpyTag (SERK-SigP_SpyTag) in RFC[25] N-Part
This part encodes the SpyTag [http://parts.igem.org/Part:BBa_K1159201 BBa_K1159201] with a N-terminal signal peptide for secretion (SERK receptor signal peptide from ''Physcomitrella patens''). 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.
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.
19 AA Linker with N-terminal Strep-tag II and C-terminal TEV cleavage site in RFC[25]
This part encodes a 19 amino acids long linker with a N-terminal ''Strep''-tag II and a C-terminal cleavage site for the TEV Protease. For further protein fusions this part is flanked by RFC[25} pre- and suffix.
TMD of SERK Receptor from Physcomitrella patens with C-terminal GFP fusion in RFC[25]
This part codes for the Transmembrane Domain of SERK Receptor from Physcomitrella patens'' with a C-terminal (intracellular) GFPmut1 fusion; SERK TMD and GFPmut1 are seperated by a middle long linker. Still, a signal peptide is needed for membrane integration. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
Signal Peptide of Ig Kappa chain from Mus musculus in RFC[25] N-part
This part codes for the signal peptide of the Ig Kappa chain from ''Mus musculus''. 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.
Splitted and engineered N-terminal FbaB for isopeptide bound formation (SpyCatcher) in RFC[25]
This part codes for a protein that recognizes and forms a covalent isopeptide bound to a oligopeptide. That oligopeptide can be found under BBa_K1159201. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
Reactive Lys for isopeptide forming reaction
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''
Splitted and engineered C-terminal FbaB for isopeptide bound formation (SpyTag) in RFC[25]
This part codes for a oligopeptide that is recognized and bound by a covalent isopeptide bound to a protein, called SpyCatcher. That catcher protein for this oligopeptdie can be found under BBa_K1159200. This part is flanked by RFC[25] pre- and suffix for further protein fusions.
Reactive Asp for isopeptide forming reaction
Oligos synthesized by sigma. Hybridized by PCR.
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. We used this part to connect our protein domains with our lipocalin domains, for our project the universal endonuclease.The sequence produced the aminoacids Gly-Gly-Ser-Gly-Gly-Gly-Ser-Gly .
This part codes for GFPmut1 which is flanked by RFC[25] pre- and suffix.
Conversion of the iGEM parts registry to SBOL2.1