pJ204-loxP-100RPU.2-ETN-BGHpA-cHS4-100RPU.2-ETN-lox2272
(Plasmid #154834)

• Purpose: Donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE), expressing two copies of etanercept
• Depositing Lab: Lise Marie Grav
• Publication: Sergeeva et al ACS Synth Biol. 2020 Aug 24. doi: 10.1021/acssynbio.0c00322.

Backbone

• Vector backbone: pJ204
• Backbone manufacturer: DNA2.0
• Backbone size w/o insert (bp): 2703
• Modifications to backbone: Amplified for USER cloning
• Vector type: Mammalian Expression, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: ETN
• Alt name: Etanercept
• Insert Size (bp): 1473
• Promoter: 100RPU.2

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Gene/Insert 2

• Gene/Insert name: ETN
• Alt name: Etanercept
• Insert Size (bp): 1473
• Promoter: 100RPU.2

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: unknown

Resource Information

• Supplemental Documents:
  • pl0688_cled.gb

Terms and Licenses

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

Depositor Comments

This is a donor plasmid for Cre/lox-based recombinase-mediated cassette exchange (RMCE) using poly(A)-trap.

Promoter 100RPU.2 was initially described in: Brown et al. 2017 "In silico design of context-responsive mammalian promoters with user-defined functionality." DOI: 10.1093/nar/gkx768

A patent application has been filed by third parties on the promoter 100RPU.2 used in this plasmid, permission for deposition of plasmids containing the promoter sequence has been provided by AstraZeneca and The University of Sheffield.

Etanercept (Enbrel) is a commercial biopharmaceutical.

The plasmid was assembled by USER cloning.