Last data update: 24 January 2024 16:39 CET
Plasmid name: pDG2i-hCARD14-FLAG-puro (LMBP 9868)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p9868.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human caspase recruitment domain family member 14 cDNA (CARD14, BIMP2, CARMA2, PSORS2) FLAG epitope tag; C-terminal Reverse tetracycline-responsive transcriptional activator (rtTA); variant 3 (rtTA3) |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE); minimal promoter with tet responsive element (CMVTRE) 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 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) Puromycin (puro) |
| Replicon: | Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) Phage f1 origin |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pDG2i-Flag puro; pENTR3C-hCARD14 |
| Further information: | The plasmid was constructed via Gateway LR recombination between pENTR3C-hCARD14 and pDG2i-Flag puro. This is a lentiviral Gateway expression vector, with the tet-inducible minimal human CMV-IE promotor and rtTA3 on the same plasmid. 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. The nucleotide sequence of the full length human CARD14 coding sequence corresponds with the Genbank accession number NM_024110.4. The nucleotide sequence of the pDG2i-Flag puro vector corresponds with the EMBL Nucleotide Sequence Database accession number LT009444.1. Other name of the plasmid is pDG2i-Flag puro_CARD14 FL. |
| EMBL Accession number: | NM_024110.4, view at GenBank LT009444.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 05/12/2016 |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | - |
| Plasmid reference: | Afonina et al., EMBO Rep. 17 (2016), 914-927 [PMID: 27113748] [DOI: 10.15252/embr.201642109] |
| Related plasmid reference: | De Groote et al., BioTechniques 60 (2016), 252-259 [PMID: 27177818] [DOI: 10.2144/000114417] |
| Restricted distribution: | - 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: | 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: |
pDG2i-hCARD14-FLAG-puro (LMBP 9868) is available at BCCM/GeneCorner. - and was published in Afonina et al., 2016. |
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.