Team:KU Leuven/Parts
BLADEN
Parts
Table of parts
| Number | Short description | Long description |
| BBa_K3807000 | enCas9-PolI3M-TBD | Variant of the EvolvR complex consisting of enhanced Cas9 enzyme fused to DNA polymerase I variant 3M with a theoredoxin binding domain. |
| BBa_K3807001 | enCas9-PolI3M | Variant of the EvolvR complex consisting of enhanced Cas9 enzyme fused to DNA polymerase I variant 3M. |
| BBa_K3807002 | enCas9-PolI5M | Variant of the EvolvR complex consisting of enhanced Cas9 enzyme fused to DNA polymerase I variant 5M |
| BBa_K3807003 | codon harmonized enCas9-PolI5M | Variant of the EvolvR complex consisting of enhanced Cas9 enzyme fused to DNA polymerase I variant 5M, codon harmonized for expression in Nicotiniana tabacum |
| BBa_K3807004 | ΔsfGFP | superfolder green fluorescent protein (GFP) with a nonsense mutation at position 93 that turns a tyrosine-coding TAT codon into a TAG stop-codon |
| BBa_K3807005 | ΔsfGFP gRNA | enCas9 gRNA scaffold that introduces a nick at postion 281 of sfGFP gene |
| BBa_K3807006 | gRNA molecular ruler 1 | enCas9 gRNA scaffold that introduces a nick at postion 327 of sfGFP gene |
| BBa_K3807007 | gRNA molecular ruler 2 | enCas9 gRNA scaffold that introduces a nick at postion 376 of the sfGFP gene |
| BBa_K3807008 | gRNA molecular ruler 3 | enCas9 gRNA scaffold that introduces a nick at postion 417 of sfGFP gene |
| BBa_K3807009 | gRNA molecular ruler 4 | enCas9 gRNA scaffold that introduces a nick at postion 472 of sfGFP gene |
| BBa_K3807010 | gRNA molecular ruler 5 | enCas9 gRNA scaffold that introduces a nick at postion 528 of sfGFP gene |
| BBa_K3807011 | gRNA molecular ruler 6 | enCas9 gRNA scaffold that introduces a nick at postion 582 of sfGFP gene |
| BBa_K3807012 | gRNA molecular ruler 7 | enCas9 gRNA scaffold that introduces a nick at postion 648 of sfGFP gene |
| BBa_K3807013 | gRNA molecular ruler 8 | enCas9 gRNA scaffold that introduces a nick at postion 693 of sfGFP gene |
| BBa_K3807014 | gRNA molecular ruler 9 | enCas9 gRNA scaffold that introduces a nick at postion 740 of sfGFP gene |
| BBa_K3807015 | gRNA molecular ruler 10 | enCas9 gRNA scaffold that introduces a nick at postion 794 of sfGFP gene |
| BBa_K3807016 | gRNA scaffold with riboswitch protospacer 1 | gRNA scaffold targeting the theophylline riboswitch (BBa_K598005). The protospacer targets the 5' end of the riboswitch so the downstream sequence gets mutated. |
| BBa_K3807017 | sfGFP | Superfolder green fluorescent protein serving as a reporter for protein expression. In our experiment, this gene is fused to a TetA gene with a flexible linker to serve as a selection platform for our SELEX process. |
| BBa_K3807018 | Riboswitch + selection device | Riboswitch device to be inserted in a low-copy number plasmid. It includes an indicable promoter (BBa_I0500), theophylline riboswitch (BBa_K598005) and TetA (BBa_J31007)-sfGFP fusion with flexible linker. This device allows for the positive and negative selection of protein expression (through the TetA gene). |
| BBa_K3807019 | pALiCE01 | Vector for cell-free protein expression |
| BBa_K3807020 | CamV35SP | A strong constitutive promoter derived from cauliflower mosaic virus commonly used in plants |
| BBa_K3807021 | NLS N7 | Commonly used in plants N-terminal nuclear localisation signal |
| BBa_K3807022 | P2A-mCherry | Self-cleaving 2A peptide fused to and mCherry with NLS. Serves as transformation marker in plants |
| BBa_K3807023 | CamV35ST | Commonly used in plants terminator derived from sauliflower mosaic virus |
| BBa_K3807024 | SacB | SacB encodes the enzyme levansucrase from Bacillus subtilis that converts sucrose into a toxic metabolite in gram-negative bacteria. Can be used as negative selection at cloning sites. |
| BBa_K3807025 | pGGK | Standard plant Golden Gate assembly backbone carrying T-dna borders, kanamycin resistance gene expressed in plats and spectinomycin resistance expressed in bacteria |
| BBa_K3807026 | GS gRNA expression cassette 1 | protospacer targeting BY-2 glutamine synthetase (GS) gene at the BASTA binding site in the 12th exon merged with enCas9 gRNA body and regulated by AtU6-26 promoter |
| BBa_K3807027 | GS gRNA expression cassette 2 | protospacer targeting BY-2 glutamine synthetase (GS) gene at the BASTA binding site in the 11th exon merged with enCas9 gRNA body and regulated by AtU6-26 promoter |
| BBa_K3807028 | neg control gRNA expression cassette | protospacer that does not have a binding site in the BY-2 nicotiana tabacum genome merged with enCas9 gRNA body and regulated by AtU6-26 promoter |
| BBa_K3807029 | plant enCas9-PolI5M expression cassette | EvolvR enCas9-PolI5M fused with N-7 NLS at the N-terminus, followed by P2A self-cleaving peptide and fluorescent protein mCherry wiht NLS as transformation marker. Expression is regulated by CamV35 promoter and terminator |
| BBa_K3807030 | plant enCas9-PolI3MTBD expression cassette | EvolvR enCas9-PolI3M-TBD fused with N-7 NLS at the N-terminus, followed by P2A self-cleaving peptide and fluorescent protein mCherry wiht NLS as transformation marker. Expression is regulated by CamV35 promoter and terminator |
| BBa_K3807033 | plant enCas9-PolI3M expression cassette | EvolvR enCas9-PolI3M fused with N-7 NLS at the N-terminus, followed by P2A self-cleaving peptide and fluorescent protein mCherry wiht NLS as transformation marker. Expression is regulated by CamV35 promoter and terminator |
| BBa_K3807032 | pGGK-p35S-NLS-N7-enCas9-PolI5M-P2A-mCherry-sacB | pGG plasmid containing Evolvr enCas9-PolI5M expression cassette (BBa_K380707029) followed by a SacB promoter and SacB gene flanked by AarI restriction sites so that it can be easily substituted by a custom gRNA expression cassette |
| BBa_K3807033 | pGGK-p35S-NLS-N7-enCas9-PolI3M-TBD-P2A-mCherry-sacB | pGG plasmid containing Evolvr enCas9-PolI3M-TBD expression cassette (BBa_K380707030) followed by a SacB promoter and SacB gene flanked by AarI restriction sites so that it can be easily substituted by a custom gRNA expression cassette |
| BBa_K3807034 | pGGK-p35S-NLS-N7-enCas9-PolI3M-P2A-mCherry-sacB | pGG plasmid containing Evolvr enCas9-PolI3M expression cassette (BBa_K380707031) followed by a SacB promoter and SacB gene flanked by AarI restriction sites so that it can be easily substituted by a custom gRNA expression cassette |
| BBa_K3807035 | gRNA scaffold with riboswitch protospacer 2 | gRNA scaffold targeting the theophylline riboswitch (BBa_K598005). The protospacer targets the 3' end of the riboswitch so the upstream sequence gets mutated. |
| BBa_K3807036 | gRNA scaffold with riboswitch protospacer 3 | gRNA scaffold targeting the theophylline riboswitch (BBa_K598005). The protospacer targets the 5' end of the TetA gene directly downstream of the riboswitch. Mutations will be introduced in the upstream sequence, which covers the riboswitch sequence. |