RNA motif plasmid cloning backbone
(Plasmid #107253)

• Purpose: RNA motif plasmid with BoxB sites. Digest with BsmBI and insert RNA motif of interest. Stuffer size 1.8kb
• Depositing Lab: Paul Khavari
• Publication: Ramanathan et al Nat Methods. 2018 Feb 5. pii: nmeth.4601. doi: 10.1038/nmeth.4601.

Backbone

• Vector backbone: pLEX
• Vector type: Mammalian Expression, Lentiviral

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

• Gene/Insert name: RNA motif plasmid cloning backbone
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBI (destroyed during cloning)
• 3′ cloning site: BsmBI (destroyed during cloning)
• 5′ sequencing primer: CAT GGT CCT GCT GGA GTT CGT G

Resource Information

• Supplemental Documents:
  • RNA_motif_plasmif with Stuffer.gb
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

Digest plasmid with BsmBI and ligate RNA motif of interest. The RNA motif of interest would be in the 3UTR and flanked by BoxB motifs.

Oligo cloning can be performed similarly to sgRNA cloning. Primers to order:
Primers (5'->3')
F GCTT <RNA motif of interest>
R AGCT <Reverse complement RNA motif of interest>

Example to clone in motif IRE motif
IRE motif: TAACACATTATCGGGAGCAGTGTCTTCCATAATGTATAA
Primers to order
F: GCTT TAACACATTATCGGGAGCAGTGTCTTCCATAATGTATAA
R: AGCT TTATACATTATGGAAGACACTGCTCCCGATAATGTGTTA