pTcI39-switch
(Plasmid #195557)

• Purpose: Temperature state switch expressing low-temperature mWasabi and high-temperature mRFP1
• Depositing Lab: Mikhail Shapiro
• Publication: Xiong et al ACS Synth Biol. 2022 Jul 15;11(7):2518-2522. doi: 10.1021/acssynbio.2c00093. Epub 2022 Jun 15.

Backbone

• Vector backbone: pETDuet-1
• Backbone manufacturer: Novagen
• Modifications to backbone: Replaced T7 Promoter #1 with pR/pL Promoter. Replaced T7 Promoter #2 with LacI Promoter.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: mRFP1
• Species: Synthetic
• Insert Size (bp): 678
• GenBank ID: CAH64892

Gene/Insert 2

• Gene/Insert name: mWasabi
• Species: Synthetic
• Insert Size (bp): 747
• GenBank ID: ABW74902.1
• Tags / Fusion Proteins:
  • AANDENYADAS degradation tag (C terminal on insert)
  • 6x His (N terminal on insert)

Gene/Insert 3

• Gene/Insert name: TcI39
• Alt name: TcI
• Alt name: Lambda Repressor (temperature sensitive)
• Alt name: CI857
• Species: Bacteriophage Lambda
• Insert Size (bp): 714
• Mutation: A67T

Resource Information

• Supplemental Documents:
  • pTcI39-state-switch.gb
• A portion of this plasmid was derived from a plasmid made by: mRFP1 was obtained from the pTU1-A-RFP plasmid5, a gift from Paul Freemont (Addgene plasmid # 72939 ; http://n2t.net/addgene:72939 ; RRID:Addgene_72939). mWasabi and TcI39 were derived from our previous work: Piraner et al Nat Chem Biol. 2017 Jan;13(1):75-80. doi: 10.1038/nchembio.2233. Epub 2016 Nov 14.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry