pLenti_EF1α-NES-Kinprola_PKA_T/A-mEGFP_bGH-PA term_EF1α_FLAG-SV40NLS-Cas9-NLS-T2A-BSD-WPRE-SV40
(Plasmid
#233372)
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PurposeEF1α driven co-expression of the inactive PKA activity recorder negative control with T/A mutation Kinprola_PKA_T/A fused to mEGFP and Cas9 expression through lentivirus transduction
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 233372 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepLenti
- Backbone size w/o insert (bp) 7028
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Vector typeLentiviral
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Selectable markersBlasticidin
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature30°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert 1
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Gene/Insert nameNES-Kinprola_PKA_T/A-mEGFP
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SpeciesSynthetic
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Insert Size (bp)2277
- Promoter EF1α
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Tags
/ Fusion Proteins
- NES (N terminal on insert)
- mEGFP (C terminal on insert)
Cloning Information for Gene/Insert 1
- Cloning method Gibson Cloning
- 5′ sequencing primer Unknown (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert nameFLAG-SV40NLS-Cas9-NLS-T2A-BSD
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SpeciesSynthetic
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Insert Size (bp)4695
- Promoter EF1α
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Tag
/ Fusion Protein
- FLAG (N terminal on insert)
Cloning Information for Gene/Insert 2
- Cloning method Gibson Cloning
- 5′ sequencing primer Unknown (Common Sequencing Primers)
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/10.1101/2024.09.11.611894 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:
pLenti_EF1α-NES-Kinprola_PKA_T/A-mEGFP_bGH-PA term_EF1α_FLAG-SV40NLS-Cas9-NLS-T2A-BSD-WPRE-SV40 was a gift from Kai Johnsson (Addgene plasmid # 233372 ; http://n2t.net/addgene:233372 ; RRID:Addgene_233372) -
For your References section:
Molecular recording of cellular protein kinase activity with chemical labeling. Sun D, Wang Ng S, Zheng Y, Xie S, Schwan N, Breuer P, Hoffmann DC, Michel J, Azorin DD, Boonekamp KE, Winkler F, Wick W, Boutros M, Li Y, Johnsson K. bioRxiv 2024.09.11.611894 10.1101/2024.09.11.611894