pBAD-LSSA12
(Plasmid #186546)

• Purpose: LSSA12 fluorescent protein (ex/em = 398/511 nm): bacterial expression plasmid
• Depositing Lab: Gregory Petsko
• Publication: Campbell et al Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 10.1038/s41592-022-01660-7.

Backbone

• Vector backbone: pBAD/His-B
• Backbone manufacturer: Invitrogen
• 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: LSSA12
• Species: Synthetic
• Insert Size (bp): 720
• Mutation: hfYFP-G65S/Y203I/Q204E/E222D/R223F
• Promoter: araB
• Tag / Fusion Protein:
  • 6xHis (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-F (ATGCCATAGCATTTTTATCCA)
• 3′ sequencing primer: pBAD-R (GATTTAATCTGTATCAGG)

Terms and Licenses

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

Depositor Comments

398 nm excitation, 511 nm emission. Backbone is identical to that of Addgene #37129 (several amino acids between the FP insert and His-tag are removed).