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Purpose(Empty Backbone) Binary vector derived from pRGE31. Deliver sgRNA and Cas9 into plant. by agrobacterium mediated transformation.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 51295 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCAMBIA1300
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Backbone manufacturerCAMBIA
- Backbone size (bp) 8958
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Modifications to backbone1. Removed Bsa I sites. 2. Inserted the U3-gRNA-35S-Cas9 cassette from pRGE31.
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Vector typePlant Expression, CRISPR ; plant expression
- Promoter Rice snoRNA U3 and dual 35S promoter
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Selectable markersHygromycin
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsFor agrobacterium strains like EHA105, growth temperature is 28 C.
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Copy numberHigh Copy
Cloning Information
- Cloning method Restriction Enzyme
- 5′ sequencing primer M13R(-48)
- 3′ sequencing primer M13F(-20) (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byThe Cas9 gene fragment was from pX330.
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References
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
The full plasmid sequence was assembled, the XmaI site is not present.
Please use the reference links above to access a published protocol for using plasmids pRGE31, pRGEB31, pRGE32 and pRGEB32 as well as a CRISPR design software tool.
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.
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For your Materials & Methods section:
pRGEB31 was a gift from Yinong Yang (Addgene plasmid # 51295 ; http://n2t.net/addgene:51295 ; RRID:Addgene_51295) -
For your References section:
RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System. Xie K, Yang Y. Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17. 10.1093/mp/sst119 PubMed 23956122