pTrc99S-ssDsbA-EPA-DNNNS-DQNRT
(Plasmid #128404)

• Purpose: Periplasmic expression of P. aeruginosa exotoxin A with DNNNS and DQNRT internal glycosylation sites in E. coli
• Depositing Lab: Michael Jewett
• Publication: Stark et al Sci Adv. 2021 Feb 3;7(6). pii: 7/6/eabe9444. doi: 10.1126/sciadv.abe9444. Print 2021 Feb.

Backbone

• Vector backbone: pTrc99S
• Backbone size w/o insert (bp): 4535
• Total vector size (bp): 6442
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin and Streptomycin, 50 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Exotoxin A
• Species: P. aeruginosa
• Insert Size (bp): 1906
• Mutation: 242DNNNS246 and 384DQNRT388 engineered glycosylation sites
• Entrez Gene: toxA (a.k.a. PA1148)
• Promoter: trc
• Tags / Fusion Proteins:
  • 6xHis tag (C terminal on insert)
  • DsbA signal sequence for periplasmic translocation (N terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GACAATTAATCATCCGGCTCG
• 3′ sequencing primer: GATTTAATCTGTATCAGG

Resource Information

• Supplemental Documents:
  • pTrc99S-ssDsbA-EPA-DNNNS-DQNRT.gb
• A portion of this plasmid was derived from a plasmid made by: Exotoxin A with internal DNNNS and DQNRT glycosylation sites was originally described by Cuccui, et al. doi.org/10.1098/rsob.130002

Terms and Licenses

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