pJ204-loxP-100RPU.2-coEPO-lox2272
(Plasmid
#154828)
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PurposeDonor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE), expressing one copy of erythropoietin
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 154828 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepJ204
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Backbone manufacturerDNA2.0
- Backbone size w/o insert (bp) 2703
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Modifications to backboneAmplified for USER cloning
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Vector typeMammalian Expression, Cre/Lox
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 numberUnknown
Gene/Insert
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Gene/Insert namecoEPO
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Alt nameErythropoietin
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SpeciesH. sapiens (human)
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Insert Size (bp)582
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MutationCodon-optimized (Cricetulus griseus)
- Promoter 100RPU.2
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer unknown (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.
Depositor Comments
This is a donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE) using poly(A)-trap.
Promoter 100RPU.2 was initially described in: Brown et al. 2017 "In silico design of context-responsive mammalian promoters with user-defined functionality." DOI: 10.1093/nar/gkx768
A patent application has been filed by third parties on the promoter 100RPU.2 used in this plasmid, permission for deposition of plasmids containing the promoter sequence has been provided by AstraZeneca and The University of Sheffield.
The plasmid was assembled by USER cloning.
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:
pJ204-loxP-100RPU.2-coEPO-lox2272 was a gift from Lise Marie Grav (Addgene plasmid # 154828 ; http://n2t.net/addgene:154828 ; RRID:Addgene_154828) -
For your References section:
Multi-copy targeted integration for accelerated development of high-producing CHO cells. Sergeeva D, Lee GM, Nielsen LK, Grav LM. ACS Synth Biol. 2020 Aug 24. doi: 10.1021/acssynbio.0c00322. 10.1021/acssynbio.0c00322 PubMed 32835482