pTBL210
(Plasmid
#126451)
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PurposeExpresses Cas9-XTEN-TdT in mammalian cells.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 126451 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3.1
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5523
- Total vector size (bp) 11238
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Modifications to backboneAdded a Kozak consensus sequence downstream of the NheI site. Switched neomycin resistance to hygromycin.
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Vector typeMammalian Expression
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Selectable markersHygromycin
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 nameCas9-XTEN-TdT
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Alt nameterminal deoxynucleotidyl transferase
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Alt nameDNA nucleotidylexotransferase
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Alt nameCas9
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SpeciesH. sapiens (human); Streptococcus pyogenes
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Insert Size (bp)5715
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MutationSee depositor comments below
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GenBank IDNM_004088.3
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Entrez GeneDNTT (a.k.a. TDT)
- Promoter CMV
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer CMV Forward
- 3′ sequencing primer BGH Reverse (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byGeorge Church; Cas9 is from Addgene plasmid 41815.
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
XTEN was used as a linker in a Cas9 fusion protein by Komor et al., 2016: doi 10.1038/nature17946.
Please Note: Plasmid contains a S9Y mutation in HygR. It is not known how this mutation affects hygromycin function. Please visit https://doi.org/10.1101/639120 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:
pTBL210 was a gift from Chang Liu (Addgene plasmid # 126451 ; http://n2t.net/addgene:126451 ; RRID:Addgene_126451) -
For your References section:
Lineage tracing and analog recording in mammalian cells by single-site DNA writing. Loveless TB, Grotts JH, Schechter MW, Forouzmand E, Carlson CK, Agahi BS, Liang G, Ficht M, Liu B, Xie X, Liu CC. Nat Chem Biol. 2021 Jun;17(6):739-747. doi: 10.1038/s41589-021-00769-8. Epub 2021 Mar 22. 10.1038/s41589-021-00769-8 PubMed 33753928