Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: John Lazar
Date created: 2016-07-14 11:00:00
Date modified: 2016-10-15 12:29:42
mRFP+gRNA Template (for saCas9)
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K2019001_sequence (Version 1) |
Description
Part BBa_K2019000 expresses crispr associated protein 9 extracted from staphylococcus aureus (saCas9) in E. Coli. saCas9 is much smaller than the typical cas9 extracted form the genome of streptococcus thermophilus (spCas9), as it the gene that encodes for saCas9 is ~1000bp shorter than that of spCas9. Furthermore, saCas9 has been shown to cut just as efficiently as spCas9 while also offering a more specific PAM sequence of NNGRRT. The his tag (his6) was attached to saCas9 to allow one to able to run a FPLC test.Notes
We linearized a backbone with the tet promoter and then used gibson assembly to attach two gblocks with the Cas9 sequence.Source
Tet promoter was used from the. SaCas9 was extracted from the genomic sequence of staphylococcus aureus and then codon optimized for E. Coli.| Sequence Annotation | Location | Component / Role(s) |
| BBa_J04450 BBa_J23119 gRNA Template gRNA Scaffolding | 1,1068 1069,1104 1104,1160 1161,1243 | sequence_feature feature/dna feature/promoter promoter sequence_feature feature/dna feature/dna sequence_feature |