pCJ208
(Plasmid
#162681)
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PurposeLidded variant of PETase from Ideonella sakaiensis 201-F6 (Genbank GAP38373.1) removing a seven-residue loop of PETase (Trp185:Phe191, PETase numbering) and replacing it with Gly251:Thr472 from MHETase; codon optimized for expression in E. coli K12, with C-terminal His tag.
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 162681 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET-21b(+)
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Backbone manufacturerEMD Biosciences
- Backbone size w/o insert (bp) 5367
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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)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namePETase from Ideonella sakaiensis 201-F6 (Genbank GAP38373.1) incorporating the MHETase lid
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SpeciesIdeonella sakaiensis 201-F6
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MutationReplacement of W185-F191 with G251-T472 from MHETase; codon optimized for expression in E. coli K12
- Promoter T7/lac
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Tag
/ Fusion Protein
- His (C terminal on backbone)
Cloning Information
- Cloning method Gibson Cloning
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:
pCJ208 was a gift from Gregg Beckham (Addgene plasmid # 162681 ; http://n2t.net/addgene:162681 ; RRID:Addgene_162681) -
For your References section:
Characterization and engineering of a two-enzyme system for plastics depolymerization. Knott BC, Erickson E, Allen MD, Gado JE, Graham R, Kearns FL, Pardo I, Topuzlu E, Anderson JJ, Austin HP, Dominick G, Johnson CW, Rorrer NA, Szostkiewicz CJ, Copie V, Payne CM, Woodcock HL, Donohoe BS, Beckham GT, McGeehan JE. Proc Natl Acad Sci U S A. 2020 Oct 13;117(41):25476-25485. doi: 10.1073/pnas.2006753117. Epub 2020 Sep 28. 10.1073/pnas.2006753117 PubMed 32989159