pCWori cytochrome P450-BM3h 9D7
(Plasmid #61308)

• Purpose: Expresses Cytochrome P450-BM3h 9D7 with C-terminal 6-His tag
• Depositing Lab: Alan Jasanoff
• Publication: Brustad et al J Mol Biol. 2012 Sep 14;422(2):245-62. doi: 10.1016/j.jmb.2012.05.029. Epub 2012 May 30.

Backbone

• Vector backbone: pCWori
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 6365
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Transform into BL21(DE3) for expression. Cultures can be grown at 37 degC until induction, after which the cultures should be incubated at 30 degC.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Cytochrome P450-BM3 heme domain 9D7
• Alt name: BM3h
• Alt name: BM3h-9D7
• Alt name: CYP102A1
• Species: Bacillus megaterium
• Insert Size (bp): 1769
• Mutation: T268A, I263A, T438V, A328G
• GenBank ID: J04832.1 142797
• Promoter: Tac
• Tag / Fusion Protein:
  • 6xHis (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (unknown if destroyed)
• 3′ cloning site: EcoRI (unknown if destroyed)
• 5′ sequencing primer: LacI-R

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: We received this plasmid from Eric Brustad (first author of the article and the researcher who cloned the gene) as a result of the collaboration between our lab (Jasanoff Lab) and the Arnold Lab on the primary reference article.

Terms and Licenses

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