8xCTS1_deltaC55G-hutRNApro_BsmBIplch-sgRNA_14nt Buffer_delta-hutRNApro_SV40polyA
(Plasmid
#124543)
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PurposeCloning vector for tRNA-released sgRNA. BsmBI flanked placeholder for spacer cloning.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 124543 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backboneP1-EYFP-pA (Construct 5) (Addgene: 55197)
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Vector typeMammalian Expression
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 numberUnknown
Gene/Insert
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Gene/Insert namedeltaC55G-hutRNApro_BsmBIplch-sgRNA_14nt Buffer_delta-hutRNApro
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gRNA/shRNA sequenceUnknown
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SpeciesH. sapiens (human)
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Mutationd11-41 C55G, d11-41
- Promoter Minimal ADE
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Unknown (unknown if destroyed)
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.
Depositor Comments
Engineered tRNA scaffold-flanked sgRNA with a BsmBI cloning site for spacer cloning. The flanking variants show low/absent background with moderate guide release capability. Enhancer elements can be exchanged using the NheI site, and promoter/enhancer region can be changed using NcoI NheI double digestion.
Please visit https://www.biorxiv.org/content/10.1101/342485v1 for BioRxiv preprint.
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:
8xCTS1_deltaC55G-hutRNApro_BsmBIplch-sgRNA_14nt Buffer_delta-hutRNApro_SV40polyA was a gift from Tudor Fulga (Addgene plasmid # 124543 ; http://n2t.net/addgene:124543 ; RRID:Addgene_124543) -
For your References section:
Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression. Knapp DJHF, Michaels YS, Jamilly M, Ferry QRV, Barbosa H, Milne TA, Fulga TA. Nat Commun. 2019 Apr 2;10(1):1490. doi: 10.1038/s41467-019-09148-3. 10.1038/s41467-019-09148-3 PubMed 30940799