Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV51E6HmDE (LMBP 3599)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3599.gb
(View with Genome Compiler) p3599.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) cDNA; heavy chain (E6H), mutated sequence |
| Promoter: | Simian virus 40 early promoter (SV40 early) Simian virus 40 late promoter (SV40 late) |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pSV51; pSV23SE6HmDE; pPLcT73 |
| Further information: | The plasmid was constructed as follows: 1) The mutated E6H cDNA was first cloned as a HindIII-SalI fragment from pSV23SE6HmDE into pPLcT73, resulting in the intermediate construction pPLcT73E6Hm407D409E; 2) The mutated E6H cDNA was then cloned as a XbaI fragment from pPLcT73E6Hm407D409E into the XbaI opened pSV51 vector (nucleotide position 3087 and 3636). As compared to the wt E6H cDNA, the codons at position 407 (Tyr) and 409 (Lys) were replaced, encoding now 2 negatively charged residues, respectively Asp and Glu. Furthermore, a unique ClaI site as well as an extra SacI and TaqI site were introduced. The expressed heavy chain will preferentially associate with a positively charged heavy chain (e.g. pSV51E6HmRK) and form heterodimers. This plasmid is useful for the expression of bifunctional or bispecific monovalent antibodies under control of the SV40 late promoter. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727356.1. The amino acid substitutions and the introduction of the ClaI and SacI site have been verified by DNA sequence analysis at the Department of Biomedical Molecular Biology (Ghent University, Belgium). Other name of the plasmid is pSV51E6Hm407D409E. |
| EMBL Accession number: | LT727356.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 02/08/2000 |
| Sequence detail: | Nucleotide sequence of the mutated E6H cDNA:
407 409
wt E6H 5'…TAC.TTC.ATA.TAT.AGC.AAG.CTC.AAT…3'
Tyr Phe Ile Tyr Ser Lys Leu Asn
mutated E6H 5'…TAC.TTC.ATC.GAT.AGC.GAG.CTC.AAT…3'
Tyr Phe Ile Asp Ser Glu Leu Asn
ClaI SacI
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglI/SnaBI, ClaI/XbaI, EcoRI, SalI and TaqI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr K. De Sutter(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | De Sutter et al., Gene 113 (1992), 223-230 [PMID: 1572543] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12xB HB101 |
| Host reference: | Boyer and Roulland-Dussoix, J. Mol. Biol. 41 (1969), 459-472 [PMID: 4896022] |
| Related host reference: | Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Sambrook et al. (eds), Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, NY (1989) [ISSN/ISBN: 0879693096] |
| 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: |
pSV51E6HmDE (LMBP 3599) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. De Sutter . |
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.