pMM643
(Plasmid #112533)

• Purpose: pZE2-PLhrtO-lux-hrtR-RBS3-chuA - HrtR RBS variant of plasmid pMM627 (Strength 33545.5 AU), ColE1 origin, KanR
• Depositing Lab: Timothy Lu
• Publication: Mimee et al Science. 2018 May 25;360(6391):915-918. doi: 10.1126/science.aas9315.

Backbone

• Vector backbone: PZE2
• Backbone size w/o insert (bp): 2216
• Total vector size (bp): 10928
• Modifications to backbone: RBS3 (stronger HrtR RBS)
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: HrtR
• Species: Lactococcus lactis
• Insert Size (bp): 570
• Promoter: ProD

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GTC AAT ACG AGG ACG CAG AAA GGG TG

Gene/Insert 2

• Gene/Insert name: ChuA
• Species: E. coli
• Insert Size (bp): 1983
• Mutation: A17P
• Promoter: J23107

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CCG TTC GTA AGC CAT TTC CGC TC

Gene/Insert 3

• Gene/Insert name: luxCDABE
• Alt name: Bacterial luciferase operon
• Species: Photorhabdus luminescens
• Insert Size (bp): 5798
• Promoter: PL(HrtO)

Cloning Information for Gene/Insert 3

• Cloning method: Gibson Cloning
• 5′ sequencing primer: GGC AAG AAA GCC ATC CAG TTT ACT TTG C

Resource Information

• Supplemental Documents:
  • pMM643.gb

Terms and Licenses

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

Depositor Comments

Blood sensor validated in murine and swine models.

The A17P mutation in ChuA is not expected to affect plasmid function.