PurposeLevel 1 Golden Gate Cassette: sgRNA cassette targeting PM19_3 in barley
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||68254||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backbonepICH47761 (AddGene #48003)
- Total vector size (bp) 4875
Vector typePlant Expression, Synthetic Biology
Growth in Bacteria
Growth instructionsCan also be grown in Agrobacterium tumefaciens GV3101/ LBA4404. Will be low copy in this species.
Copy numberHigh Copy
Gene/Insert namePromoter_TaU6 + sgRNA_HvPM19_3
SpeciesTritucum aestivum and synthetic
Insert Size (bp)507
- Cloning method Restriction Enzyme
- 5′ cloning site BsaI (destroyed during cloning)
- 3′ cloning site BsaI (destroyed during cloning)
- 5′ sequencing primer GAACCCTGTGGTTGGCATGCACATAC
- 3′ sequencing primer CTGGTGGCAGGATATATTGTGGTG (Common Sequencing Primers)
Target sequence: GGCTGGCGTTGGTCGTAACA
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pICSL11058 was a gift from Nicola Patron (Addgene plasmid # 68254 ; http://n2t.net/addgene:68254 ; RRID:Addgene_68254)
For your References section:Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease. Lawrenson T, Shorinola O, Stacey N, Li C, Ostergaard L, Patron N, Uauy C, Harwood W. Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7. 10.1186/s13059-015-0826-7 [pii] PubMed 26616834