pET28a_T5-ARG1
(Plasmid #106476)

• Purpose: Expresses acoustic reporter genes or gas vesicles (ARG1) in Escherichia coli Nissle 1917
• Depositing Lab: Mikhail Shapiro
• Publication: Bourdeau et al Nature. 2018 Jan 3;553(7686):86-90. doi: 10.1038/nature25021.

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5189
• Total vector size (bp): 11944
• Vector type: Bacterial Expression, Synthetic Biology

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin + 2% Glucose
• Growth Temperature: 37°C
• Growth Strain(s): NEB Turbo
• Growth instructions: Following transformation, rescue in SOC broth for 1 hour at 37C before transferring the rescued cells (entire volume) into overnight growth media (LB+ 50 ug/mL Kanamycin + 2% glucose) for growth at 37C. 2% glucose is optional, but it's recommend to prevent any leaky protein expression from the constitutive T5 promoter driving the ARG gene. Usually prepare a 40% stock solution of glucose and dilute it to the final concentration of 2%.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: ARG1
• Alt name: Acoustic Reporter Gene 1
• Alt name: Hybrid Gas Vesicle gene cluster
• Species: Anabaena flos-aquae and Bacillus megaterium
• Insert Size (bp): 6755
• Promoter: T5

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: custom primers
• 3′ sequencing primer: T7 terminator_Reverse

Resource Information

• Supplemental Documents:
  • pET28a_T5-ARG1.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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