Last data update: 24 January 2024 16:39 CET
Plasmid name: pROSA26-COIN-DV-mVEGFA-593 (LMBP 8185)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p8185.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse microRNA 30 (Mir30, Mirn30, mmu-mir-30); fragment Short hairpin RNA sequence targeting mouse vascular endothelial growth factor A (Vegfa, Vegf, VEGF120, VEGF164, VEGF188, VEGF-A, VPF) 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; pEntry attR2-IRES-eGFP-attL3; pUgly-mVEGFA-593; pEntry-attL4-rtTA-IRES-Puro-pA-attR1 |
| Further information: | The plasmid was constructed via multisite Gateway recombination between pROSA26-COIN-DV, pEntry-attL4-rtTA-IRES-Puro-pA-attR1, pUgly-mVEGFA-593 and pEntry attR2-IRES-eGFP-attL3. The siRNA encoded by this plasmid starts at nucleotide position 593 of the mouse Vegfa coding sequence. This multisite Gateway Expression vector 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. This plasmid is part of a first generation COIN (GOF/LOF) system, in which the mouse Vegfa-593 shRNA 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 VEGF.593. |
| 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-mVEGFA-593 (LMBP 8185) 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.