pET28a_6HT-TBP6.7(P51A)
(Plasmid
#112728)
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PurposeSimilar to P46, P51 was selected and is present in all evolved TBPs. Mutating this position into Ala has a modest effect on binding.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 112728 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET28a(+)
- Backbone size w/o insert (bp) 5277
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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 name6His-TEV-TBP6.7(P51A)
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Insert Size (bp)359
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MutationP51A
- Promoter T7
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Tag
/ Fusion Protein
- 6His-TEV (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NcoI (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer T7 promoter
- 3′ sequencing primer T7 terminator (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.
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:
pET28a_6HT-TBP6.7(P51A) was a gift from Joseph Wedekind (Addgene plasmid # 112728 ; http://n2t.net/addgene:112728 ; RRID:Addgene_112728) -
For your References section:
Structure of HIV TAR in complex with a Lab-Evolved RRM provides insight into duplex RNA recognition and synthesis of a constrained peptide that impairs transcription. Belashov IA, Crawford DW, Cavender CE, Dai P, Beardslee PC, Mathews DH, Pentelute BL, McNaughton BR, Wedekind JE. Nucleic Acids Res. 2018 Jul 27;46(13):6401-6415. doi: 10.1093/nar/gky529. 10.1093/nar/gky529 PubMed 29961805