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PurposePlasmid for IPTG-inducible expression and purification of protein A-Hia5-6xHis (pA-Hia5) for antibody directed DNA (adenine) methylation (as in DiMeLo-seq)
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Depositing Lab
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 174372 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
This material is available to academics and nonprofits only.
Backbone
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Vector backbonepET
- Total vector size (bp) 6534
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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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Growth instructionsFor protein expression, use T7 Express lysY competent E. coli cells (NEB #C3010I) See protocol: dx.doi.org/10.17504/protocols.io.bv82n9ye
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameprotein A - hia5
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Insert Size (bp)1500
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Tags
/ Fusion Proteins
- Protein A (N terminal on insert)
- 6xHis (C terminal on backbone)
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
Protocol for purification and expression can be found at: dx.doi.org/10.17504/protocols.io.bv82n9ye
Application for DiMeLo-seq is described in: doi.org/10.1101/2021.07.06.451383
Protocol was adapted and modified from: doi.org/10.1126/science.aaz1646
How to cite this plasmid
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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:
pET-pA-Hia5 was a gift from Aaron Straight (Addgene plasmid # 174372 ; http://n2t.net/addgene:174372 ; RRID:Addgene_174372) -
For your References section:
DiMeLo-seq: a long-read, single-molecule method for mapping protein-DNA interactions genome wide. Altemose N, Maslan A, Smith OK, Sundararajan K, Brown RR, Mishra R, Detweiler AM, Neff N, Miga KH, Straight AF, Streets A. Nat Methods. 2022 Jun;19(6):711-723. doi: 10.1038/s41592-022-01475-6. Epub 2022 Apr 8. 10.1038/s41592-022-01475-6 PubMed 35396487
Map uploaded by the depositor.
