pLGB13
(Plasmid #126618)

• Purpose: (Empty Backbone) Suicide vector for allelic replacement in Bacteroides, erythromycin selection and aTC-inducible ss-Bfe1 counterselection
• Depositing Lab: Laurie Comstock
• Publication: Garcia-Bayona et al mBio. 2019 Aug 13;10(4). pii: mBio.01762-19. doi: 10.1128/mBio.01762-19.

Backbone

• Vector backbone: pNBU2_erm-TetR-P1T_DP-GH023
• Modifications to backbone: 1) Chemically synthesized cassette (Genscript) containing a periplasmic localization signal (first 63 bp of BT_4676 from B. thetaiotaomicron VPI-5482) fused to the bfe1 gene (BF638R_1988) of B. fragilis 638R. (Lim et al., 2017) 2) Using NEBuilder assembly, this cassette was cloned into the NcoI site of pNBU2_erm-TetR-P1T_DP-GH023. 3) The resulting plasmid was used as template for PCR , to remove the attN2 site and the integrase gene, and the resulting product was assembled using NEBuilder.
• Vector type: allellic exchange in bacteria
• Selectable markers: erythromycin (Bacteroidales)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): S17-lambda pir
• Copy number: Low Copy

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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