pET28-his-3C_Clu3B6p
(Plasmid
#100093)
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Purposeexpresses easily crystallizable engineered 14-3-3 sigma variant fused to the HSPB6 phosphorylatable peptide
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 100093 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET28-his-3C
- Backbone size w/o insert (bp) 5369
- Total vector size (bp) 6149
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Vector typeBacterial Expression
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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert name14-3-3 sigma human
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Alt name14-3-3 sigma fusion
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SpeciesH. sapiens (human)
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Insert Size (bp)780
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Mutation75-77 residues changed to AAA to facilitate crystallization
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Entrez GeneSFN (a.k.a. YWHAS)
- Promoter T7
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Tags
/ Fusion Proteins
- HSPB6 peptide (C terminal on insert)
- His6 (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NcoI (not destroyed)
- 3′ cloning site XhoI (not destroyed)
- 5′ sequencing primer T7 promoter
- 3′ sequencing primer T7 terminator (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
The 3C site was introduced immediately prior to 14-3-3 sequence to facilitate purification by subtractive IMAC.
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:
pET28-his-3C_Clu3B6p was a gift from Nikolai Sluchanko (Addgene plasmid # 100093 ; http://n2t.net/addgene:100093 ; RRID:Addgene_100093) -
For your References section:
Chimeric 14-3-3 proteins for unraveling interactions with intrinsically disordered partners. Sluchanko NN, Tugaeva KV, Greive SJ, Antson AA. Sci Rep. 2017 Sep 20;7(1):12014. doi: 10.1038/s41598-017-12214-9. 10.1038/s41598-017-12214-9 [pii] PubMed 28931924