ECFP CD
(Plasmid #207414)

• Purpose: Mammalian-cell-based plasmid system that expresses ECFP (a non R-loop-forming transcript) upon dox-induction, which will further cause co-directional transcription-replication conflict (CD TRC).
• Depositing Lab: Karlene Cimprich
• Publication: Hamperl et al Cell. 2017 Aug 10;170(4):774-786.e19. doi: 10.1016/j.cell.2017.07.043.

Backbone

• Vector backbone: pCEP4 Mammalian Expression Vector
• Backbone manufacturer: Thermo Fisher
• Backbone size w/o insert (bp): 10400
• Total vector size (bp): 10822
• Modifications to backbone: 1) Replacement of pCMV with tight-Tet-ON promoter; 2) Switching orientation of tight-Tet-ON promoter by SalI restriction enzyme digestion and religation; 3) Cloning of ECFP or mAIRN sequence downstream of tight-Tet-ON promoter
• Vector type: Mammalian Expression, Bacterial Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: ECFP
• Species: Synthetic; Aequorea victoria
• Insert Size (bp): 717

Resource Information

• Supplemental Documents:
  • pSH27.gb

Terms and Licenses

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

Depositor Comments

Please Note: Addgene NGS is not able to fully resolve the EBNA1 feature within the plasmid due to the repetitive nature of the sequence. Please refer to the depositor's expected sequence for this region of the plasmid.