pHelper_TS_V2_ampR
(Plasmid #214467)

• Purpose: Helper plasmid for ORBIT in E. coli. Expresses RecT for oligo recombineering and Bxb-1 for pInt incorporation. Includes temperature sensitive origin for plasmid curing.
• Depositing Lab: Scott Saunders
• Publication: Saunders et al bioRxiv 2023

Backbone

• Vector backbone: pKD46
• Vector type: Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Plasmid has temperature sensitive replication and is lost at 37°C or 42°C. It must be grown at 30°C to replicate.
• Copy number: Low Copy

Gene/Insert

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

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: NA
• 3′ sequencing primer: NA

Resource Information

• Supplemental Documents:
  • phelper_ts_v2_ampr.gb
• A portion of this plasmid was derived from a plasmid made by: George Church (Addgene plasmid # 138474: pORTMAGE-Ec1), Kenan Murphy (Addgene plasmid # 108320: pKM461), and Coli Genetic Stock Center (CGSC #7739: pKD46).

Terms and Licenses

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