Last data update: 24 January 2024 16:39 CET
Plasmid name: pT2-CMV-3xHA-hTERT-Neo (LMBP 9746)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9746.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human telomerase reverse transcriptase cDNA (TERT, TRT, CMM9, EST2, DKCA2, DKCB4, GeneID 7015) Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal Modified hyperactive sleeping beauty transposon arms |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Simian virus 40 early promoter (SV40 early) Phage T7 gene 10 promoter (T7g10) Phage T3 promoter Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Neomycin (neo; G418) Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pCDNA-3xHA-hTERT; pT2/BH |
| Further information: | The plasmid was constructed by isolating a cassette, expressing the HA-tagged human TERT coding sequence under control of the hCMV-IE promoter and the BGH terminator, followed by a neomycin resistance marker under the control of the SV40 promoter and terminator, from pCDNA-3xHA-hTERT as a HincII fragment, and cloning it into the EcoRV-opened pT2/BH transposon plasmid. The TERT-Neo cassette is flanked by modified hyperactive sleeping beauty transposon arms consisting of IR/DRs (= inverted repeats containing short direct repeats). Sleeping Beauty (SB) transposon-based transfection is a two-component system consisting of a transposase and a transposon containing IR/DR sequences that result in precise integration into a TA-dinucleotide site. The purpose of this plasmid is to enable high-throughput stable integration for immortalization by co-transfection with a sleeping beauty transposase (SB100X). This plasmid can be co-transfected with pT2-SVT, which immortalizes cells through a different pathway. |
| EMBL Accession number: | - |
| Latest sequence update: | 28/10/2019 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr R. Beyaert(1) (2) and Dr J. Staal(1) (2). (1) Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5α |
| Host reference: | Focus 8 (1986), 9 |
| Related host reference: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pT2-CMV-3xHA-hTERT-Neo (LMBP 9746) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and Dr J. Staal . |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.