Last data update: 24 January 2024 16:39 CET
Plasmid name: pDG1i-hE-cad-eGFP-V5 blast (LMBP 7954)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p7954.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human cadherin 1 cDNA (CDH1, E-cad, UVO, CD324, GeneID 999) Aequorea victoria green fluorescent protein DNA (GFP); enhanced red-shifted variant (EGFP), C-terminal V5 epitope; C-terminal |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE); minimal promoter with tet operator (CMVT) Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR) Simian virus 40 early promoter (SV40 early) Escherichia coli lac operon promoter Phage SP6 promoter Phage T3 promoter Phage T7 gene 10 promoter (T7g10) |
| Ribosome binding site: |
Internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV) polyprotein gene |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 3' LTR); modified HIV-1 3' LTR with deleted U3 region (ΔU3/3' LTR) |
| Selection marker: | Ampicillin (amp) Blasticidin (bsd) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pDG1i-eGFP-V5 blast |
| Further information: | The plasmid was constructed by cloning the human CDH1 coding sequence into the pDG1i-eGFP-V5 blast vector. This is a lentiviral Gateway expression vector, with tet-inducible human CMV-IE promotor. The U3 region of the HIV-1 3' LTR is deleted (ΔU3 or dU3) and facilitates self-inactivation of the HIV-1 5' LTR after transduction to enhance the biosafety of the vector. The element also contains a polyadenylation signal for transcription termination and polyadenylation of mRNA in transduced cells. The vector contains the psi packaging signal, allowing viral packaging, and the rev response element (RRE) from HIV-1, permitting Rev-dependent nuclear export of unspliced viral mRNA. Because the risk for recombination after each subcultivation is high, this plasmid is only available under the format of isolated plasmid DNA. Plasmid integrity should be checked via PvuII restriction digest after each subcultivation.Some leakage expression of human CDH1 may be detected. Other name of the plasmid is pDG1 Ecadh-EGFP. |
| EMBL Accession number: | - |
| Latest sequence update: | 24/06/2015 |
| 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 P. De Groote(1) (2) and Prof. Dr W. Declercq(1) (2). (1) Inflammation Research Center, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | 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 in E. coli; L2 in mammalian cells |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pDG1i-hE-cad-eGFP-V5 blast (LMBP 7954) is available at BCCM/GeneCorner. This plasmid was deposited by Dr P. De Groote and Prof. Dr W. Declercq . |
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.