Last data update: 24 January 2024 16:39 CET
Plasmid name: pT2-EF1a-hHOIL1-R165A-Blast (LMBP 9675)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | not available |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human RANBP2-type and C3HC4-type zinc finger containing 1 cDNA (RBCK1, HOIL1, RBCK2, RNF54, UBCE7IP3, XAP4, ZRANB4, GeneID 10616); mutated sequence Modified hyperactive sleeping beauty transposon arms |
| Promoter: | Human elongation factor 1α promoter (EF1α) Phage T3 promoter Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) |
| Selection marker: | Blasticidin (bsd) Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pENTR-P4P1R-EF1a; pENTR221-hHOIL-1L-R165A; pENTR-C-BGH-Blast; pdT2-BH-L4L3 |
| Further information: | The plasmid was constructed via multisite Gateway LR recombination, combining the human EF1a promoter from pENTR-P4P1R-EF1a, the human RBCK1 mutant coding sequence from pENTR221-hHOIL-1L-R165A and the BGH terminator with blasticidin resistance gene from pENTR-C-BGH-Blast into the pdT2-BH-L4L3 vector. The human RBCK1 expression cassette is flanked by modified hyperactive sleeping beauty transposon arms consisting of IR/DRs (= inverted repeats containing short direct repeats). The human RBCK1 coding sequence contains an R165A mutation. The purpose of this transposon plasmid is simple stable integration of the mutated human RBCK1 coding sequence in a cell by co-transfection with a sleeping beauty transposase (SB100X). 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. |
| EMBL Accession number: | - |
| Latest sequence update: | 28/02/2018 |
| 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 Dr J. Staal(1) (2) and Prof. R. Beyaert(1) (2). It was constructed by J. Meert(2)(3). (1) VIB-UGent Center for Inflammation Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium (3) BCCM/GeneCorner Plasmid Collection, 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-EF1a-hHOIL1-R165A-Blast (LMBP 9675) is available at BCCM/GeneCorner. This plasmid was deposited by Dr J. Staal and Prof. R. Beyaert . |
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.