PASTA-2_trunc-pUC19_INT-Bxb1-attV_P2A_CD19CAR_bGH-pA
(Plasmid
#247038)
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PurposeIntegration-Plasmid with Bxb1-attV (docking site) for bxb1-mediated integration of a CD19-CAR. Targeted whole plasmid insertion after maxiprep + AMPure bead purification.
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Depositing Lab
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Depositing OrganizationCharite Universitaetsmedizin Berlin
Located in Germany -
Publication
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 247038 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
Backbone
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Vector backbonetruncated pUC19
- Backbone size w/o insert (bp) 1846
- Total vector size (bp) 3912
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Modifications to backboneWe removed non-essential sequences from the pUC19 vector to generate a truncated version (tpUC19), thereby reducing the size, complexity, and bacterial DNA content of the integration plasmid.
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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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameBxb1 phage attachment site (attP) = vector attachment site (attV), CD19-CAR
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SpeciesH. sapiens (human), Synthetic; Bacteriophage Bxb1
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
This is PL#1 from our ‘one-pot’ PASTA publication. This integration-plasmid enables expression of a CD19-CAR through Bxb1-mediated in-frame insertion into any translated exon of a transcriptionally active gene (in suitable cell types such as primary human T cells). It is intended for use together with eeBxb1 mRNA, which can be generated by IVT from David Liu’s Addgene plasmid #222339. For TRAC integration, generate HDR-template from plasmid ‘PASTA-1_pUC19_HDR-Bxb1-attG_TRAC_for-Cas12a’ (Addgene #247037), for Cas12a-assisted HDR-mediated TRAC integration of the corresponding Bxb1 landing pad (attB = attG). Please visit https://doi.org/10.1101/2025.09.10.675267 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:
PASTA-2_trunc-pUC19_INT-Bxb1-attV_P2A_CD19CAR_bGH-pA was a gift from Dimitrios Wagner (Addgene plasmid # 247038 ; http://n2t.net/addgene:247038 ; RRID:Addgene_247038) -
For your References section:
Versatile and efficient non-viral integration of large transgenes in human T cells via CRISPR knock-in and engineered integrases. Kassing I, Kath J, Nitulescu A-M, Glaser V, Hartmann LM, Pu Y, Huth L, Kārkliņš R, Shaji S, Ringel AR, Pouzolles M, Stein M, Ibrahim DM, Wagner DL. bioRxiv 2025.09.10.675267 10.1101/2025.09.10.675267