pCMV5-25BoxB
(Plasmid #60817)

• Purpose: Cloning vector, 25 repeats of 19 bp BoxB sequence from bacteriophage-λ, for RNA labeling. BoxB-RNA is bound by 22 amino acid RNA binding domain of λ-anti-terminator protein N, e.g. tagged with GFP
• Depositing Lab: Maria Carmo-Fonseca
• Publication: Martin et al Cell Rep. 2013 Sep 26;4(6):1144-55. doi: 10.1016/j.celrep.2013.08.013. Epub 2013 Sep 12.

Backbone

• Vector backbone: pCMV5
• Backbone manufacturer: Dr. David Russell, UTSW
• Backbone size w/o insert (bp): 4657
• Total vector size (bp): 5639
• Vector type: Mammalian Expression, Synthetic Biology

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: 25x BoxB
• Alt name: binding site of bacteriophage λ antitermination protein N
• Alt name: boxb
• Species: Synthetic; bacteriophage λ
• Insert Size (bp): 1042
• Mutation: 25 repeats of BoxB sequence
• GenBank ID: NC_001416.1 1QFQ_A
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: hGH-PA-R

Resource Information

• A portion of this plasmid was derived from a plasmid made by: A construct with 5x boxb repeats for tethering of RNA was described in Gehring et al. Mol Cell. 2003 Apr;11(4):939-49. The authors provided us with the original published reporter gene containing 5x boxb. From this precursor the 5x boxb sequence was amplified by PCR, cloned into a new vector backbone pCMV5 and multiplied to finally contain 25x boxb sequence repeats by us.
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

the authors of Gehring et al. Mol Cell. 2003 Apr;11(4):939-49 were contacted by us and granted their permission to distribute the 25xboxb construct generated in our lab from their precursor for non-profit purpose via Addgene