Cloning and expression plasmid for one gene of interest
pUNO plasmids feature a wide choice of native and fusion genes. The gene of interest is under the control of of the strong and ubiquitous mammalian promoter, EF1α/HTLV.
pUNO plasmids can be used for in vitro or in vivo transfection experiments.
Invivogen also provides pUNO-mcs plamids, designed to serve as control and/or cloning vectors.Back to the top
pUNO1: selectable with Blasticidin in mammalian cells and E.coli
pUNO2: selectable with Zeocin™ in mammalian cells and E.coli
pUNO3: selectable with Hygromycin in mammalian cells and E.coli
pUNO4: selectable with Puromycin in mammalian cells and E.coli
• hEF1/HTLV prom is a composite promoter comprising the Elongation Factor-1α (EF-1α) core promoter and the R segment and part of the U5 sequence (R-U5’) of the Human T-Cell Leukemia Virus (HTLV) Type 1 Long Terminal Repeat. The EF-1α promoter exhibits a strong activity, higher than viral promoters and, on the contrary to the CMV promoter, yields persistent expression of the transgene in vivo. The R-U5’ has been coupled to the EF-1α core promoter to enhance stability of DNA and RNA.
• MCS or Gene:
- The multiple cloning site contains the following restriction sites:
pUNO: 5’ - SgrAI, SalI, BamHI, Eco47III, PstI, NheI - 3’
pUNO1: 5’ - Sal I, SgrA I, BamH I, Eco47 III, Nco I, Nhe I - 3’
Each restriction site is compatible with many other enzymes, increasing the cloning options.
• SV40 pan: The Simian Virus 40 late polyadenylation signal enables efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA.
• ori is a minimal E. coli origin of replication with the same activity as the longer Ori.
• SV40 prom: The Simian Virus 40 promoter allows the expression of the blasticidin resistance gene in mammalian cells.
• CMV enh/prom: The human cytomegalovirus immediate-early gene 1 promoter/enhancer was originally isolated from the Towne strain and was found to be stronger than any other viral promoters.
• Resistance gene:
The resistance gene is driven by the SV40 promoter in tandem with the bacterial EM7 promoter to allow antibiotic selection in both mammalian cells and E. coli bacteria.
- bsr: The bsr gene from Bacillus cereus encodes a deaminase that confers resistance to the antibiotic Blasticidin S.
- Sh ble: Resistance to Zeocin™ is conferred by the Sh ble gene from Streptoalloteichus hindustanus which inactivates Zeocin™ upon binding to the antibiotic.
- hph: Resistance to Hygromycin B is conferred by the hph gene from E. coli which encodes a phosphotransferase.
- pac: The pac gene from Streptomyces sp. encodes a puromycin N-acetyl-transferase that confers resistance to Puromycin.
• βGlo pAn: The human beta-globin 3’UTR and polyadenylation sequence allows efficient arrest of the transgene transcription.
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- 20 µg of lyophilized DNA.