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PurposeBsaI-based cloning of SpCas9 gRNA guide sequence, position 3/13/23 in the array
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 80786 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepMA15
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Backbone manufacturerLifetechnologies
- Backbone size w/o insert (bp) 2380
- Total vector size (bp) 2789
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Modifications to backboneno
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Vector typeMammalian Expression, CRISPR
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameCRISPR gRNA expression cassette (for SpCas9)
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gRNA/shRNA sequenceempty (BbsI)
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SpeciesSynthetic
- Promoter U6
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Tag
/ Fusion Protein
- na
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site BsrGI (not destroyed)
- 5′ sequencing primer TAATACGACTCACTATAGG (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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.
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:
pMA-SpCas9-g3 was a gift from Yonglun Luo (Addgene plasmid # 80786 ; http://n2t.net/addgene:80786 ; RRID:Addgene_80786) -
For your References section:
Golden Gate Assembly of CRISPR gRNA expression array for simultaneously targeting multiple genes. Vad-Nielsen J, Lin L, Bolund L, Nielsen AL, Luo Y. Cell Mol Life Sci. 2016 May 13. 10.1007/s00018-016-2271-5 [pii] PubMed 27178736