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PurposeMammalian expression of SpCas9 PE7 prime editor with P2A-human MLH1dn (codon optimized)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 222997 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepCMV
- Backbone size w/o insert (bp) 3402
- Total vector size (bp) 12795
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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)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namePE7-P2A-hMLH1dn
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Alt namePEmax-La(SSB)1-194-P2A-hMLH1dn
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SpeciesH. sapiens (human), Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus; La(SSB)1-194 is from Homo sapiens
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Insert Size (bp)9393
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MutationDeletion of residues 754-756 in MLH1 as described in https://doi.org/10.1016/j.cell.2021.09.018
- Promoter CMV
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Tags
/ Fusion Proteins
- SV40 bpNLS (N terminal on insert)
- SV40 bpNLS (C terminal on insert)
- c-Myc NLS (C terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer CMV Forward, T7
- 3′ sequencing primer BGHR (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:
pCMV-PE7-P2A-hMLH1dn was a gift from Brittany Adamson (Addgene plasmid # 222997 ; http://n2t.net/addgene:222997 ; RRID:Addgene_222997) -
For your References section:
Improving prime editing with an endogenous small RNA-binding protein. Yan J, Oyler-Castrillo P, Ravisankar P, Ward CC, Levesque S, Jing Y, Simpson D, Zhao A, Li H, Yan W, Goudy L, Schmidt R, Solley SC, Gilbert LA, Chan MM, Bauer DE, Marson A, Parsons LR, Adamson B. Nature. 2024 Apr 3. doi: 10.1038/s41586-024-07259-6. 10.1038/s41586-024-07259-6 PubMed 38570691