Last data update: 24 January 2024 16:39 CET
Plasmid name: pVitro2-mGS-TL1A-Ig (LMBP 8496)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p8496.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse glutamine synthetase cDNA (GS) Mouse tumor necrosis factor ligand member 15 cDNA (TL1A); fragment Mouse immunoglobulin γ1 cDNA (IgG1); κ-chain leader sequence Mouse immunoglobulin γ1 cDNA (IgG1); hinge region and CH2 and CH3 domains of the constant portion of the heavy chain (Fc) |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only Simian virus 40 early promoter (SV40 early); enhancer only Phage T7 gene 10 promoter (T7g10) Human ferritin heavy subunit promoter (FerH) Human ferritin light subunit promoter (FerL) |
| Ribosome binding site: |
Internal ribosome entry site (IRES) of the foot and mouth disease virus (FMDV) |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Hygromycin (hyg) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pVitro2-hygro-mcs |
| Further information: | The plasmid was created by first PCR amplifying the mouse IgG κ-chain leader sequence and the 5'′ end of the cDNA encoding the Fc portion of mouse IgG1, containing the hinge, CH2, and CH3 regions, and ligating them into a TOPO cloning vector. Mouse IgG1 was mutated to replace the three cysteines of the hinge portion to serines. Next, the extracellular domain of the mouse TL1A cDNA, containing amino acids 68 to 252, was PCR amplified and ligated to the 3'′ end of the mouse IgG1 Fc fragment cDNA. The cDNA construct containing mouse IgG κ-chain leader sequence, mouse IgG1 Fc fragment and mouse TL1A fragment was cloned as a SgraI/NheI fragment into the second multiple cloning site of pVitro2-mGS, which had the mouse GS cDNA inserted as a MluI/SalI fragment in the first multiple cloning site. The nucleotide sequence of the mouse IgG1 Fc fragment corresponds with the EMBL Nucleotide Sequence Database accession number AB538879.1. The nucleotide sequence of the mouse TL1A cDNA corresponds with the Genbank accession number NM_177371.3. The nucleotide sequence of the mouse GS cDNA corresponds with the Genbank accession number NM_008131.4. The nucleotide sequence of the FMDV IRES corresponds with the EMBL Nucleotide Sequence Database accession number AF274010.1 except for a missing C between nucleotide position 15 and 16, a C instead of T in position 86, a G instead of A in position 446 and a C instead of A in position 453 of the IRES sequence. The nucleotide sequence of the 3' UTR of human EF1a corresponds with the Genbank accession number NM_001402.5 except for a G instead of T in position 558 of the 3' UTR sequence. Other name of the plasmid is mIgG1-mTL1A mGS pVitro2 mcs. |
| EMBL Accession number: | AB538879.1, view at EMBL, GenBank, DDBJ NM_177371.3, view at GenBank NM_008131.4, view at GenBank AF274010.1, view at EMBL, GenBank, DDBJ NM_001402.5, view at GenBank |
| Latest sequence update: | 01/07/2013 |
| 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 E. Podack(1). (1) Sheila and David Fuente Program in Cancer Biology, University of Miami Miller School of Medicine, Miami, USA |
| Plasmid reference: | Khan et al., J. Immunol. 190 (2013), 1540-1550 [PMID: 23319737] |
| 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 + hygromycin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pVitro2-mGS-TL1A-Ig (LMBP 8496) is available at BCCM/GeneCorner. This plasmid was deposited by Dr E. Podack and was published in Khan et al., 2013. |
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.