pET151-Vps4(101-437)-Hcp1 K321A/R325A
(Plasmid
#89287)
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PurposeBacterial expression of S. cerevisiae Vps4 AAA ATPase cassette fused to P. aeruginosa Hcp1
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 89287 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET151
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5700
- Total vector size (bp) 7300
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Vector typeBacterial Expression
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 numberLow Copy
Gene/Insert
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Gene/Insert nameVacuolar protein sorting-associated protein 4
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Alt nameVps4
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SpeciesS. cerevisiae (budding yeast)
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Insert Size (bp)1554
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Mutationdeleted amino acids 1-100, mutated Lys-321 and Arg-325 to Ala
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GenBank IDNM_001184270
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Entrez GeneVPS4 (a.k.a. YPR173C, CSC1, DID6, END13, GRD13, VPL4, VPT10)
- Promoter T7
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Tags
/ Fusion Proteins
- 6xHis-V5-TEV (N terminal on backbone)
- Hcp1 (C terminal on insert)
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer TAATACGACTCACTATAGG
- 3′ sequencing primer GCTAGTTATTGCTCAGCGG (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.
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:
pET151-Vps4(101-437)-Hcp1 K321A/R325A was a gift from Chris Hill (Addgene plasmid # 89287 ; http://n2t.net/addgene:89287 ; RRID:Addgene_89287) -
For your References section:
Structural Basis of Protein Translocation by the Vps4-Vta1 AAA ATPase. Monroe N, Han H, Shen PS, Sundquist WI, Hill CP. Elife. 2017 Apr 5;6. pii: e24487. doi: 10.7554/eLife.24487. 10.7554/eLife.24487 PubMed 28379137