Last data update: 24 January 2024 16:39 CET
Plasmid name: pSCUDMFE6L2 (LMBP 3205)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3205.gb
(View with Genome Compiler) p3205.txt p3205.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Saccharomyces cerevisiae α-mating factor 1 gene (MFα1); prepro secretion signal sequence (ppMF) Mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) cDNA; light chain (E6L) missing the leader signal sequence |
| Promoter: | Saccharomyces cerevisiae iso-1-cytochrome C promoter (CYC1) Saccharomyces cerevisiae galactokinase promoter (GAL1) Saccharomyces cerevisiae translation elongation factor 1α promoter (TEF1α) |
| Ribosome binding site: |
- |
| Terminator: | Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) |
| Selection marker: | Ampicillin (amp) Bleomycin (bleo; zeomycin (zeo; Zeocin); phleomycin (phleo)) Methotrexate (MTX) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli Saccharomyces cerevisiae; integrative |
| Parental clone: | pUDHIL208; pSP64ble3; pSCUDT2MFE6L |
| Further information: | This plasmid was constructed as follows: 1) the intermediate plasmid pSCUDMFE6L1 was derived from pUDHIL208 by replacing the BamHI fragment, containing the human interleukin 2 expression cassette, by the BamHI fragment of pSCUDT2MFE6L, containing the light chain E6L of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) expression cassette; 2) the HindIII fragment of this intermediate vector, containing the URA3 selection marker, was then substituted by the HindIII fragment of pSP64ble3, containing the bleomycin selection marker. Expression of the mouse dihydrofolate reductase gene (mDHFR) confers resistance to methotrexate. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727450.1. Since the nucleotide sequence between the S. cerevisiae GAL1 promoter and the S. cerevisiae TY1 transposable element could not be completely traced back, there is uncertainty as to the cutting frequency of the restriction enzymes, except for the sites that were analysed in the authenticity test. |
| EMBL Accession number: | LT727450.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 23/08/2001 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI/HindIII, Eco88I, NdeI/Psp5II, PdmI, PvuI and PvuII. As compared to the compiled nucleotide sequence, the restriction site analysis pattern revealed that there are approximately 150 extra nucleotides located between the PvuI site at position 6433 and the Eco88I site at position 7315, most probably in the unknown sequence region, and that there is an extra NdeI or Psp5II site in the bleomycin expression cassette since the theoretical expected 1196 bp Psp5II fragment resulted on gel in fragments of approximately 1000 bp and 200 bp. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by W. Lammerant(1) and Prof. Dr R. Contreras(1). (1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Mumberg et al., Nucleic Acids Res. 22 (1994), 5767-5768 [PMID: 7838736] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSCUDMFE6L2 (LMBP 3205) is available at BCCM/GeneCorner. This plasmid was deposited by W. Lammerant and Prof. Dr R. Contreras. |
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.