pInt_attP1_tsXis_sacB_kanR
(Plasmid #205302)

• Purpose: (Empty Backbone) Integrating plasmid for ORBIT in E. coli with self excising factor (Xis) and sacB for counter selection. Used for markerless modifications.
• Depositing Lab: Scott Saunders
• Publication: Saunders et al bioRxiv 2023

Backbone

• Vector backbone: pKD4
• Vector type: Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): BW25141
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: None

Resource Information

• Supplemental Documents:
  • pint_attp1_tsxis_sacb_kanr.gb
• A portion of this plasmid was derived from a plasmid made by: pKM496 (Addgene #109301), pKD4 (CGSC #7632), pKM461 (Addgene #108320), and pBad-Xis/Int Reset Flipper (Addgene #38207)

Terms and Licenses

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

Depositor Comments

This plasmid is pInt_attP1_kanR with the sacB gene (sucrose sensitivity) and the Bxb-1 excisionase (Xis) under temperature sensitive regulation (induced at 37C - 42C). This is used for markerless modifications (leaves attB scar). Normal (uninduced) grow temperature is 30C. Please visit https://doi.org/10.1101/2023.06.28.546561 for bioRxiv preprint. Depositing lab uses BW25141 strain for growth (https://cgsc.biology.yale.edu/Strain.php?ID=64690)