pSJ111
(Plasmid #78707)

• Purpose: Modular vector for constitutive secreted amylase reporter expression
• Depositing Lab: Wiep Klaas Smits
• Publication: Oliveira Paiva et al ACS Synth Biol. 2016 Jun 23.

Backbone

• Vector backbone: pRPF185
• Backbone manufacturer: Fagan and Fairweather
• Backbone size w/o insert (bp): 6378
• Total vector size (bp): 8437
• Modifications to backbone: KpnI,SacI to remove Ptet, replaced with Pveg SacI, BamHI to remove gusA, replaced with amyEopt
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol, 25 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): MC1061
• Growth instructions: For selection in C. difficile, use thiamphenicol
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: amyEopt
• Alt name: secreted alpha-amylase
• Species: Synthetic
• Insert Size (bp): 1962
• Promoter: Pveg

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: NF_793 (CACCTCCTTTTTGACTTTAAGCCTACGAATACC)
• 3′ sequencing primer: NF_794 (CACCGACGAGCAAGGCAAGACCG)

Resource Information

• Supplemental Documents:
  • pSJ111 Genbank file

Terms and Licenses

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