pBAD/His-D-iLight
(Plasmid #170270)

• Purpose: Expression of iLight with 6xHis N-terminal tag in bacteria.
• Depositing Lab: Vladislav Verkhusha
• Publication: Kaberniuk et al Nat Commun. 2021 Jun 23;12(1):3859. doi: 10.1038/s41467-021-24212-7.

Backbone

• Vector backbone: Modified pBAD/His-B
• Backbone manufacturer: Life Technologies-Invitrogen (partially modified)
• Backbone size w/o insert (bp): 3999
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: iLight
• Species: Idiomarina sp. A28L
• Insert Size (bp): 1608
• Mutation: See Comments
• Promoter: araBAD
• Tag / Fusion Protein:
  • 6xHis tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pBAD Forward
• 3′ sequencing primer: pBAD Reverse

Resource Information

• Supplemental Documents:
  • pBADHis-D-iLight.gb

Terms and Licenses

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

Depositor Comments

To design iLight for expression in bacteria, a full-length bacterial phytochrome IsPadC from Idiomarina sp. A28L was used as an original template. First, diguanylyl cyclase effector in IsPadC was removed, then following mutations were introduced: I68F, H80Q, A86T, R90S S242C, E274K, R295H I360V, and L464V.