Last data update: 24 January 2024 16:39 CET
Plasmid name: pROSA26-COIN-DV-Ins/Ins-p53-1224 (LMBP 8181)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner non-core plasmid |
Depositor's sequence: | p8181.gb |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Mouse microRNA 30 (Mir30, Mirn30, mmu-mir-30); fragment Short hairpin RNA sequence targeting mouse transformation related protein 53 (Trp53, p44, p53) Mouse gene trap ROSA 26 gene (Gtrosa26, Gt(ROSA)26Sor, beta geo, Gtrgeo26, R26, ROSA26, Thumpd3as1, GeneID 14910); 5' UTR and 3' UTR |
Promoter: | Mouse phosphoglycerate kinase 1 promoter (PGK1) 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) Simian virus 40 polyadenylation signal (SV40 polyA) |
Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
Host range: | Escherichia coli Mammalian cells |
Parental clone: | pROSA26-COIN-DV-Ins/Ins; pEntry attR2-IRES-eGFP-attL3; pUgly-p53-1224; pEntry-attL4-rtTA-IRES-Puro-pA-Ins/Ins-attR1 |
Further information: | The plasmid was constructed via a multisite Gateway recombination between pROSA26-COIN-DV-Ins/Ins, pEntry-attL4-rtTA-IRES-Puro-pA-Ins/Ins-attR1, pUgly-p53-1224 and pEntry attR2-IRES-eGFP-attL3. The siRNA encoded by this plasmid starts at nucleotide position 1224 of the p53 coding sequence. The plasmid contains: - two bacteriophage P1 Cre recombinase target sites (loxP sites) - a splice acceptor (SA) for linking the rtTA-IRES-puromycin to the endogenous ROSA26 promoter - two adjacent copies of the chicken β-globin 5' DNase I hypersensitive site 4 (5'HS4) insulator core sequence. Insulators are DNA sequences which possess the ability to protect expressing genes from inappropriate signals emanating from their surrounding environment by acting as barriers that prevent the advance of nearby condensed chromatin that may otherwise silence expression. This multisite Gateway expression vector is part of a second generation, insulated COIN (GOF/LOF) system, in which the miR30-shRNA cassette is to be targeted to the mouse ROSA26 locus via homologous recombination and can be expressed conditionally and inducibly. The G4 ROSALUC ES cells (LMBP 10507CB) are available at BCCM/GeneCorner as well. Other name of the plasmid is pROSA26 COIN DV Insulated p53.1224. |
EMBL Accession number: | - |
Latest sequence update: | 01/12/2016 |
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 J. Haigh(1). (1) Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, Australia |
Plasmid reference: | PhD thesis Lieven Haenebalcke |
Restricted distribution: | - VIB/BCCM MTA - The depositor will be informed of the customer's identity upon release of a sample outside the depositor's department or outside the departments in which BCCM/GeneCorner is embedded, namely UGent-DBMB and VIB-IRC. |
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: | 28°C |
Biosafety level: | L1 |
Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pROSA26-COIN-DV-Ins/Ins-p53-1224 (LMBP 8181) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr J. Haigh and was published in Lieven Haenebalcke, . |
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.