pET-21b(+)-Is-PETase-W185A
(Plasmid #112204)

• Purpose: pET-21b(+) based plasmid for expression of PETase from Ideonella sakaiensis 201-F6 (Genbank GAP38373.1) with W185A mutations, codon optimized for expression in E. coli K12
• Depositing Labs: Gregg Beckham, Christopher Johnson
• Publication: Austin et al Proc Natl Acad Sci U S A. 2018 May 8;115(19):E4350-E4357. doi: 10.1073/pnas.1718804115. Epub 2018 Apr 17.

Backbone

• Vector backbone: pET-21b(+)
• Backbone manufacturer: Novagen/Millipore Sigma
• Backbone size w/o insert (bp): 5367
• Total vector size (bp): 6237
• Modifications to backbone: His tag: CACCACCATCACCACCAC
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: PETase gene from Ideonella sakaiensis 201-F6 with W185A mutation, codon optimized for expression in E. coli K12
• Species: Ideonella sakaiensis
• Mutation: W185A
• GenBank ID: GAP38373.1
• Promoter: N/A
• Tag / Fusion Protein:
  • 6XHIS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Resource Information

• Supplemental Documents:
  • pCJ162 A1_m4 Ideonella sakaiensis PETase gene W185A in pET-21b(+).gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry