pK-bes
(Plasmid #254055)

• Purpose: (Empty Backbone) Suicide vector for allelic exchange with sucrose-based negative selection via sacB gene. Derived from pKNOCK-bla-ermGb and pCVD442.
• Depositing Lab: Juliane Bubeck-Wardenburg
• Publication: An ubiquitous antibacterial toxin from human gut bacteria engenders neonatal colonization advantage (unpublished)

Backbone

• Vector backbone: pKNOCK-bla-ermGb
• Backbone manufacturer: Jeffrey Gordon Lab. WUSTL. Koropatkin et al., Structure, 2008. DOI: 10.1016/j.str.2008.03.017
• Backbone size (bp): 3040
• Modifications to backbone: rpoD promoter (gene bt1311 from Bacteroides thetaiotaomicron, amplified from pEV1) and sacB coding sequence (amplified from pCVD442) were inserted in KpnI-digested pKNOCK-bla-ermGb using Gibson Assembly. A Multiple Cloning Site allows insert cloning for allelic exchange purposes..
• Vector type: Allelic exchange in Bacteroides
• Promoter: rpoD (for sacB)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Pir1
• Growth instructions: Contains ermG gene for erythromycin selection in Bacteroides recipients. Contains sacB for negative selection of merodiploid clones.
• Copy number: Low Copy

Cloning Information

• Cloning method: Gibson Cloning

Terms and Licenses

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

Depositor Comments

Publication for pKNOCK-bla-ermGb backbone: DOI: 10.1016/j.str.2008.03.017. pCVD442 template for sacB obtained through Addgene (Plasmid #11074).