pLIGFP_W220F
(Plasmid #46772)

• Purpose: Protein overexpression with the modified LacIWF repressor
• Depositing Lab: Florian Hollfelder
• Publication: Gatti-Lafranconi et al Microb Cell Fact. 2013 Jul 8;12:67. doi: 10.1186/1475-2859-12-67.

Backbone

• Vector backbone: pET22b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5493
• Total vector size (bp): 6093
• Modifications to backbone: Position 220 in LacI is mutated from tryptophan to phenylalanine

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: LacI_W220F
• Alt name: LacIWF
• Alt name: LacI
• Insert Size (bp): 1083
• Mutation: Tryptophan 220 replaced by Phenylalanine
• Entrez Gene: lacI (a.k.a. b0345, ECK0342)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: cgccattcgatggtgtcc
• 3′ sequencing primer: aggctctcaagggcatcgg

Terms and Licenses

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

Depositor Comments

Use BL21 for expression. Growth temperature depends on the protein being over-expressed and is not affected by the modified repressor LacIWF.