pEXP5-NT/6xHis mCherry F30 2xdBroccoli
(Plasmid #169233)

• Purpose: Constitutive T7 RNAP-mediated expression of 6xHis-mCherry with 2xd Broccoli RNA aptamer.
• Depositing Lab: Dora Tang
• Publication: Gonzales et al ACS Synth Biol. 2022 Jan 21;11(1):205-215. doi: 10.1021/acssynbio.1c00376. Epub 2022 Jan 4.

Backbone

• Vector backbone: pEXP5-NT
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2665
• Total vector size (bp): 3691
• Modifications to backbone: None
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: 6xHis mCherry F30-2xdBroccoli
• Species: Synthetic; Discosoma sp.
• Insert Size (bp): 1023
• Promoter: T7 promoter

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: None
• 3′ sequencing primer: GGTTATTGTCTCATGAGCG

Resource Information

• Supplemental Documents:
  • pEXP5-NT_6xHis mCherry F30-2xdBroccoli.gb

Terms and Licenses

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

Depositor Comments

Plasmid used for monitoring transcription and translation dynamics. Broccoli aptamer (2xdBroccoli) provides a fluorescent signal with DFHBI for mRNA level and mCherry provides fluorescent signal for protein level. Design and sequence of the Broccoli aptamer is from Filonov et al. (2015, doi: 10.1016/j.chembiol.2015.04.018).