pDB-hPER2-Halo-CD4-bla
(Plasmid
#179451)
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PurposeDonor vector to knock in Halotag C-terminal to human PER2 gene
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 179451 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepDB, derived from pUC57
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Backbone manufacturernone
- Backbone size w/o insert (bp) 8200
- Total vector size (bp) 9187
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Modifications to backboneregions homologous to the hPER2 genomic region (surrounding STOP codon) flank insert+positive selection cassette (mCerulean-P2A-blaR). Silent mutations to the hPER2 genomic sequence were inserted to avoid donor vector targeting by Cas9/sgRNA. Negative Selection cassette expresses hCD4. Should be used together with Cas9/sgRNA plasmids #179454, 179455, 179456
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Vector typeCRISPR
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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 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 nameHaloTag
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Alt nameHalo7
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SpeciesSynthetic
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Insert Size (bp)887
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Entrez GeneHalo7 (a.k.a. Halo7)
- Promoter None
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Tag
/ Fusion Protein
- His/Flag (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site AgeI (not destroyed)
- 3′ cloning site SpeI (not destroyed)
- 5′ sequencing primer --
- 3′ sequencing primer -- (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byhCD4 was cloned from Addgene #35712
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.
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:
pDB-hPER2-Halo-CD4-bla was a gift from Achim Kramer (Addgene plasmid # 179451 ; http://n2t.net/addgene:179451 ; RRID:Addgene_179451) -
For your References section:
Live-cell imaging of circadian clock protein dynamics in CRISPR-generated knock-in cells. Gabriel CH, Del Olmo M, Zehtabian A, Jager M, Reischl S, van Dijk H, Ulbricht C, Rakhymzhan A, Korte T, Koller B, Grudziecki A, Maier B, Herrmann A, Niesner R, Zemojtel T, Ewers H, Granada AE, Herzel H, Kramer A. Nat Commun. 2021 Jun 18;12(1):3796. doi: 10.1038/s41467-021-24086-9. 10.1038/s41467-021-24086-9 PubMed 34145278