Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Isobel Holden
Date created: 2016-10-13 11:00:00
Date modified: 2016-10-19 08:03:43
sgRNA containing two golden gate adapters
| Types | DnaRegion |
| Roles | mature_transcript_region RNA |
| Sequences | BBa_K1994021_sequence (Version 1) |
Description
sgRNA with Bsa 1 Golden Gate adaptorsNotes
To simplify the traditional gRNA comprised of a crRNA containing a dCas9 handle, and tracrRNA which binds to the PAM sequence in CRISPR, we created a joined sgRNA. As modifying the 3' end of the gRNA does not affect dCas9 binding we have used a deactivated dcas9 terminator and placed golden gate adaptable sites. The two sites are located where the complementary PAM-proximal sequence is located, allowing the distance dCas9 binds from the promoter to be adapted.The other golden gate adaptor in the position after the deactivated terminator so any RNA sequence such as an RNA binding protein handle can be inserted. This region is then followed by a real terminator for the strand.Source
Synthetic modification of crRNA and tracrRNA found in S. pyrogenes| Sequence Annotation | Location | Component / Role(s) |
| Golden Gate adaptor dCas9 handle deactivated dCas9 terminator Golden Gate adaptor aspA terminator | 1,20 21,62 63,96 97,116 116,157 | sequence_feature feature/misc stem_loop feature/stem_loop feature/stem_loop stem_loop feature/misc sequence_feature stem_loop feature/stem_loop |