mc2 puro hPGK
(Plasmid #206227)

• Purpose: (Empty Backbone) For expressing cloned exons as circular RNA. Alternatively, for expressing cDNA (if the EcoRI-XhoI restriction fragment is also removed).
• Depositing Labs: Simon Conn, Brett Stringer
• Publication: Stringer et al bioRxiv. 2023 Nov 22:2023.11.21.568171. doi: 10.1101/2023.11.21.568171. Preprint.

Backbone

• Vector backbone: pcDNA3.1(+) CircRNA Mini Vector
• Backbone manufacturer: Addgene ID 60648
• Vector type: Mammalian Expression, Synthetic Biology
• Selectable markers: Puromycin

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: None

Resource Information

• Supplemental Documents:
  • mc2 puro hPGK.gb

Terms and Licenses

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

Depositor Comments

ZKSCAN1 intron 1 (partial) & ZKSCAN1 intron 3 (partial) separated by a pair of BsmBI restriction sites. 5' Cloning Site: EcoRI (not destroyed), 3' Cloning Site: XhoI (not destroyed). The mc2 and pF mc2 series of plasmids were designed and created by Dr Brett Stringer in the laboratory of Prof Simon Conn. They are based on the original circRNA mini vector design of Liang & Wilusz, 2014. The pair of BsmBI restriction sites is the cloning site for DNA specifying the RNA sequence to be circularised. For cloning cDNA, the 5' restriction site should be either NheI, BmtI, AflII, HindIII, Acc65I, KpnI, Eco53kI, SacI, BamHI, BstXI or EcoRI and the 3' restriction site should be either XhoI, PspXI or XbaI. The neoR gene of Addgene plasmid ID 60648 also was replaced with the puroR gene from Addgene plasmid ID 98290. The CMV enhancer/promoter of Addgene plasmid ID 60648 also was replaced with the hPGK promoter from Addgene plasmid ID 96923. Please visit https://doi.org/10.1101/2023.11.21.568171 for bioRxiv preprint.