Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV71BLAHI (LMBP 2543)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p2543.gb
(View with Genome Compiler) p2543.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Ampicillin resistance gene (amp) Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region |
| Promoter: | Simian virus 40 late promoter (SV40 late) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Chloramphenicol (cam) |
| Replicon: | Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli; preferably recombination-deficient strains Mammalian cells; SV40 permissive cells |
| Parental clone: | pSV71; pMac254BLAHI |
| Further information: | The plasmid was constructed as follows: The fusion gene of ampicillin resistance and the human immunoglobulin γ3 hinge region (BLAHI) was isolated as RspXI-fragment (nucleotide positions 1644 and 2825) from pMac254BLAHI; the 5' sticky ends were filled in with Klenow DNA polymerase and XbaI linkers ('CCTCTAGAGG') were ligated. Subsequently, the BLAHI gene was cloned as an XbaI-fragment into the XbaI opened pSV71, in the expression orientation relative to the SV40 late promoter. The BlaHi fusion gene was cloned as a XbaI fragment into the XbaI opened pSV71, in the expression orientation relative to the SV40 late promoter. Due to the construction, the bacterial promoter in front of the ampicillin resistance gene was lost, so amp can't be used anymore as a selection marker. Nevertheless, the transformants grow on ampicillin selective medium. β-lactamase is also enzymatically active after expression in COS-1 cells. The ampicillin resistance gene is directly fused to the start of the first hinge region H1; translation of this fusion gene terminates 8 amino acids after the end of H4. Name of the plasmid mentioned in the reference is pSV71BlaHi. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727525.1. |
| EMBL Accession number: | LT727525.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 22/01/1997 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI/PstI, EcoRV, NcoI/PvuI, SacI/XmnI, StyI and XbaI/XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr K. De Sutter(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | De Sutter et al., Mol. Microbiol. 6 (1992), 2201-2208 [PMID: 1406260] |
| 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 + chloramphenicol (25 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSV71BLAHI (LMBP 2543) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. De Sutter and was published in De Sutter et al., 1992. |
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.