pHelper_NoMutL_V2_gentR
(Plasmid #205293)

• Purpose: Helper plasmid for ORBIT in E. coli. Expresses RecT for oligo recombineering and Bxb-1 for pInt incorporation. Has sacB for counter selection.
• Depositing Lab: Scott Saunders
• Publication: Saunders et al bioRxiv 2023

Backbone

• Vector backbone: pKM461
• Vector type: Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Gentamicin, 10 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: CspRecT / Bxb-1
• Species: Synthetic

Cloning Information

• Cloning method: Gibson Cloning

Resource Information

• Supplemental Documents:
  • phelper_nomutl_v2_gentr.gb
• A portion of this plasmid was derived from a plasmid made by: pORTMAGE-Ec1 (Addgene #138474), pKM461 (Addgene #108320), pKD46 (CGSC #7739), and pMQ30 (Shanks et al. 2006)

Terms and Licenses

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

Depositor Comments

Replicating ORBIT V2 helper plasmid w/ inducible oligo recombineering module (m-tol) and bxb-1 integrase (arabinose). SacB is present for counterselection and gentamicin is the resistance marker. V2 plasmid does not have mutL, which lowers background mutation rate. Please visit https://doi.org/10.1101/2023.06.28.546561 for bioRxiv preprint.