Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-T4DNApol (JO638)
(Plasmid
#208956)
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PurposeA variant CE1 construct with T4 DNA polymerase (-exo, activity enhanced), expressed from CMV or T7 promoters.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 208956 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
This material is available to academics and nonprofits only.
Backbone
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Vector backbonepCMV-T7-SpCas9-P2A-EGFP (RTW3027)
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Vector typeMammalian Expression, CRISPR ; in vitro transcription; T7 promoter
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 namePCV2-XTEN-nSpCas9-BPNLS-T4pol-BPNLS
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Alt nameJO638
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SpeciesSynthetic
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MutationnSpCas9(H840A), T4Pol(-exo;D189A/E191A/L412M)
- Promoter CMV and T7
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Tag
/ Fusion Protein
- BPNLS (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer oBK6928-CCAAGTCTCCACCCCATTGACG
- 3′ sequencing primer oBK219-GGGAGTGGCACCTTCCAGGGTC (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:
Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-T4DNApol (JO638) was a gift from Benjamin Kleinstiver (Addgene plasmid # 208956 ; http://n2t.net/addgene:208956 ; RRID:Addgene_208956) -
For your References section:
Click editing enables programmable genome writing using DNA polymerases and HUH endonucleases. da Silva JF, Tou CJ, King EM, Eller ML, Ma L, Rufino-Ramos D, Kleinstiver BP. bioRxiv. 2023 Sep 13:2023.09.12.557440. doi: 10.1101/2023.09.12.557440. Preprint. 10.1101/2023.09.12.557440 PubMed 37745481