pENTR-R4R3-stop
(Plasmid
#113745)
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PurposeContains three stop codons in every frame to ensure a premature stop codon when introduced into the genome via homology-directed repair. Gateway multisite entry clone.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 113745 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepDONR221-P4rP3r
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 2540
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Vector typeCRISPR ; Gateway Entry Vector
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 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 namestop cassette
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SpeciesSynthetic
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Insert Size (bp)340
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MutationAdded 3 stop codons to the original sequence; see Depositor Comments below
Cloning Information
- Cloning method Gateway Cloning
- 5′ sequencing primer None
- 3′ sequencing primer None (Common Sequencing Primers)
Resource Information
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Addgene Notes
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
Second position vector for three-way Gateway LR reaction to generate a premature stop codon at your locus of interest. Also contains a multiple cloning site for easy genotyping. This plasmid contains multiple M13F and M13R sites, due to cloning the MCS of pBluescriptSK+.
QC note: This plasmid dimerizes during growth in bacteria, which can result in digest and Sanger sequencing results that differ from the full plasmid sequence of the monomer. The depositing lab has confirmed that dimerization does not affect plasmid function.
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:
pENTR-R4R3-stop was a gift from Magdalena Bezanilla (Addgene plasmid # 113745 ; http://n2t.net/addgene:113745 ; RRID:Addgene_113745) -
For your References section:
Efficient and modular CRISPR-Cas9 vector system for Physcomitrella patens. Mallett DR, Chang M, Cheng X, Bezanilla M. Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. eCollection 2019 Sep. 10.1002/pld3.168 PubMed 31523744