pSilencer2.1-U6-T1-U6-AP3
(Plasmid #190828)

• Purpose: For mammalian expression of T1 and AP3, the RNA aptamers of ncOGT and GFP, respectively. Expression of each aptamer is driven by a U6 promoter.
• Depositing Lab: Gerald Hart
• Publication: Zhu et al Cell. 2022 Dec 30:S0092-8674(22)01529-X. doi: 10.1016/j.cell.2022.12.016.

Backbone

• Vector backbone: pSilencer2.1-U6 hygro
• Total vector size (bp): 5290
• Vector type: Mammalian Expression, Synthetic Biology, Affinity Reagent/ Antibody
• Selectable markers: Hygromycin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: T1 (an RNA aptamer of OGT)
• Species: Synthetic
• Insert Size (bp): 98
• Promoter: U6

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: T7, LKO.1 5'
• 3′ sequencing primer: 2.0Rev

Gene/Insert 2

• Gene/Insert name: AP3 (an RNA aptamer of GFP)
• Species: Synthetic
• Insert Size (bp): 84
• Promoter: U6

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: LKO.1 5'

Resource Information

• Supplemental Documents:
  • 190828_pSilencer2.1-U6-T1-U6-AP3.gbk

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

Sequence of the AP3 portion is from the article below:
Shui, B., Ozer, A., Zipfel, W., Sahu, N., Singh, A., Lis, J.T., Shi, H., and Kotlikoff, M.I. (2012). RNA aptamers that functionally interact with green fluorescent protein and its derivatives. Nucleic Acids Res 40, e39. 10.1093/nar/gkr1264.