Polymerase variant click editor (CE1) - pCMV-T7-PCV2-nCas9-M-MLV(PE2) (JO586)
(Plasmid
#217800)
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PurposeVariant CE1 construct with engineered M-MLV reverse transcriptase (RT; found in PE2), expressed from CMV or T7 promoters
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Depositing Lab
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Depositing OrganizationMassachusetts General Hospital
Located in United States of America -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 217800 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 217800-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 217800-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepCMV-T7-SpCas9-P2A-EGFP (RTW3027)
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Backbone manufacturerAddgene #139987
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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-M-MLV(PE2)-BPNLS
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Alt nameJO586
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SpeciesSynthetic
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MutationnSpCas9(H840A); PE2 M-MLV(D200N/L603W/T330P/T306K/W313F)
- Promoter CMV and T7
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Tag
/ Fusion Protein
- BPNLS (C terminal on backbone)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer oBK6928-CCAAGTCTCCACCCCATTGACG
- 3′ sequencing primer oBK219-GGGAGTGGCACCTTCCAGGGTC
- (Common Sequencing Primers)
Resource Information
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.
DNA (Catalog # 217800-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 217800-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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-M-MLV(PE2) (JO586) was a gift from Benjamin Kleinstiver (Addgene plasmid # 217800 ; http://n2t.net/addgene:217800 ; RRID:Addgene_217800) -
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
Map uploaded by the depositor.
