Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Zexu Li
Date created: 2016-09-11 11:00:00
Date modified: 2016-10-18 10:24:08
tCas9-Vp64(Prokaryotic)
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K1982006_sequence (Version 1) |
Description
The CRY2/CIBN interaction is entirely genetically encoded. The binding reverses within minutes in the dark, allowing rapid shutoff of transcription by placing samples in the dark. This fusion protein is for use in LACE(light-activated CRISPR/Cas9 effector) system, and a tCas9 fused to its N terminus. To regulate DNA transcription by blue light, the system is based on CRY2/CIBN interaction in which a light-mediated protein interaction brings together two protein (tCas9 and an activation domain VP64) . If we remove the stimulation of blue light, dark reversion of CRY2 will dissociate the interaction with CIBN and shut off transcription.tCas9 can be tagged with transcriptional activators, and targeting these dCas9 fusion proteins to the promoter region results in robust transcription activation of downstream target genes. This tCas9-based activators is the case that tCas9 fused directly to a single transcriptional activator( VP64).
Notes
mutant EcoR 1 : TGTTGGGAATTc--->TGTTGGGAATTgSource
tCas9 is from GE Share company. VP64 is synthetic construct.| Sequence Annotation | Location | Component / Role(s) |
| Prokaryotic rbs ATG tCas9 SV40 NLS VP16 VP16 VP16 VP16 Linker HA FLAG TAATAA | 1,18 19,21 22,4122 4123,4143 4144,4182 4183,4221 4222,4260 4261,4299 4300,4305 4306,4332 4339,4362 4363,4368 | feature/rbs ribosome_entry_site start_codon feature/start feature/protein CDS sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc feature/misc sequence_feature feature/misc sequence_feature feature/tag tag feature/tag tag feature/stop stop_codon |