pENTRdeltaGFPmirGE-DroshaT1.5.5
(Plasmid #61084)

• Purpose: pENTR plasmid containing a mutated GFP followed by 3 miRNA haipins: recommended as backbone.
• Depositing Lab: Patrick Salmon
• Publication: Myburgh et al Mol Ther Nucleic Acids. 2014 Oct 28;3:e207. doi: 10.1038/mtna.2014.58.

Backbone

• Vector backbone: pENTRL1L2
• Backbone size w/o insert (bp): 2718
• Total vector size (bp): 3350
• Modifications to backbone: A triple stop codon is inserted at the beginning of GFP ORF to reduce GFP translation. GFP is thus used as a spacer before the miRNAS hairpins. This plasmid contains 3 tandem hairpins that can be excised by BamHI and XbaI to insert your pet mirGE hairpin as described in our article (MTNA, 2014).
• Vector type: RNAi

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: deltaGFP
• Alt name: 3 stop codons
• Alt name: no GFP translation
• gRNA/shRNA sequence: miRNA (mirGE) triple
• Species: H. sapiens (human)

Terms and Licenses

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

Depositor Comments

From the depositor, regarding a "c" missing from the miRNA: "The mutation is likely from the PCR since the hairpin is amplified by PCR.
I would not use this clone to KD Drosha but in our case, the insert is
used as an indicator of cutting when you use BamHI and XbaI to clone
your own miRNA."