pPD3C
(Plasmid
#86971)
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PurposeExpresses PCNA3 protein fused phosphite dehydrogenase and P450cam in E. coli
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 86971 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET-15b(+)
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 5700
- Total vector size (bp) 8775
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Modifications to backboneA SapI recognition site (GAAGAGC) was modified to GTTGAGC
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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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Growth instructionsSupplement with 1% glucose is recommended to suppress leaky expression
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Copy numberLow Copy
Gene/Insert
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Gene/Insert namePCNA3 fused to phosphite dehydrogenase and P450cam
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Alt namePTDH-PCNA3-P450cam
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SpeciesSynthetic; Pseudomonas, Sulfolobus
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Insert Size (bp)3075
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MutationE175A/A176R in PTDH, N106R in PCNA3, silent mutations in P450cam
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GenBank IDAAA25760.1 WP_009988781.1
- Promoter T7 promoter
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Tag
/ Fusion Protein
- His6 tag (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NdeI (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer T7 Terminal (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byStephen Sligar, University of Illinois at Urbana-Champaign (pUS200, Addgene plasmid # 17788)
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:
pPD3C was a gift from Teruyuki Nagamune (Addgene plasmid # 86971 ; http://n2t.net/addgene:86971 ; RRID:Addgene_86971) -
For your References section:
Supramolecular protein assembly supports immobilization of a cytochrome P450 monooxygenase system as water-insoluble gel. Tan CY, Hirakawa H, Nagamune T. Sci Rep. 2015 Mar 3;5:8648. doi: 10.1038/srep08648. 10.1038/srep08648 PubMed 25733255