pH6-TEV-spTorA-GFP(C)
(Plasmid #169041)

• Purpose: GFP preceeded by the signal peptide of TorA (spTorA) with an N-terminal 6xHis tag and TEV cleavage site in pBAD24.
• Depositing Lab: Siegfried Musser
• Publication: Bageshwar et al PLoS One. 2021 Sep 9;16(9):e0256715. doi: 10.1371/journal.pone.0256715. eCollection 2021.

Backbone

• Vector backbone: pBAD24
• Backbone manufacturer: Beckwith Lab
• Backbone size w/o insert (bp): 4542
• Total vector size (bp): 5413
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: H6-spTorA-GFP
• Species: Escherichia coli and Aequorea victoria
• Insert Size (bp): 900
• Mutation: The mut3 GFP variant with 5 additional mutations (Q80R, S147H, M153T, V163A, S202D). There are also 3 silent mutations at G65 (GGT instead of GGG), Q69 (CAA instead of CAG) and P75 (CCA instead of CCG). Internal NcoI site in GFP was deleted (CCATGG to CCCTGG).
• GenBank ID: CAD6011594.1 AAB18957.1
• Promoter: araBAD
• Tags / Fusion Proteins:
  • 6xHis (N terminal on insert)
  • TEV cleavage site (N terminal on insert)
  • spTorA (signal peptide of E. coli TorA) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: PstI (not destroyed)
• 5′ sequencing primer: pBAD forward primer
• 3′ sequencing primer: pBAD reverse primer

Resource Information

• Supplemental Documents:
  • pH6-TEV-spTorA-GFP(C).gbk

Terms and Licenses

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

Depositor Comments

The spTorA-GFP coding sequence was recovered from p-spTorA-GFP-H6C, a plasmid originally reported as pTorA-GFP in Bageshwar & Musser 2007 J. Cell Biol., 179:87-99.