Lentiviral click editor (CE1) plasmid - LTR-pEF1a-PCV2-nCas9-EcKlenow (DRR921)
(Plasmid
#217798)
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PurposeLentiviral plasmid to produce the prototypical CE1 construct (PCV2-nCas9-EcKlenow), expressed from the EF1a promoter
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 217798 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonelentiCRISPR v2
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Backbone manufacturerFeng Zhang, Addgene #52961
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Vector typeMammalian Expression, Lentiviral, CRISPR
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature30°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameXTEN-nSpCas9-BPNLS-EcKlenow(-exo)-BPNLS
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Alt nameDRR921
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SpeciesSynthetic
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MutationnSpCas9(H840A); EcKlenow(-exo;D355A/E357A)
- Promoter EF1a
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Tag
/ Fusion Protein
- BPNLS (C terminal on backbone)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer oBK394-GTGATGTCGTGTACTGGCTCC
- 3′ sequencing primer oBK195-GGCGGTCCGGATCGACGGTG (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.
Depositor Comments
Please visit https://doi.org/10.1101/2023.09.12.557440 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:
Lentiviral click editor (CE1) plasmid - LTR-pEF1a-PCV2-nCas9-EcKlenow (DRR921) was a gift from Benjamin Kleinstiver (Addgene plasmid # 217798 ; http://n2t.net/addgene:217798 ; RRID:Addgene_217798) -
For your References section:
Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases. Ferreira da Silva J, Tou CJ, King EM, Eller ML, Rufino-Ramos D, Ma L, Cromwell CR, Metovic J, Benning FMC, Chao LH, Eichler FS, Kleinstiver BP. Nat Biotechnol. 2024 Jul 22. doi: 10.1038/s41587-024-02324-x. 10.1038/s41587-024-02324-x PubMed 39039307