Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV51E6HmE (LMBP 3602)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3602.gb
(View with Genome Compiler) p3602.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; pSV23SE6HmE; pPLcT73 |
| Further information: | The plasmid was constructed as follows: 1) The mutated E6H cDNA was first cloned as a HindIII-SalI fragment from pSV23SE6HmE into pPLcT73, resulting in the intermediate construction pPLcT73E6Hm394E; 2) The mutated E6H cDNA was then cloned as a XbaI fragment from pPLcT73E6Hm394E into the XbaI opened pSV51 vector (nucleotide position 3087 and 3636). As compared to the wt E6H cDNA, the codons at position 394 (Thr) and 395 (Ala) were replaced, respectively by a Glu and a Leu codon. Furthermore, an extra SacI site and an extra AluI site were introduced. Due to the introduction of an acidic amino acid at position 394, the expressed heavy chain will preferentially associate with a heavy chain containing a basic amino acid (e.g. pSV51E6HmK) 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 LT727359.1. The amino acid substitutions and the introduction of the 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 pSV51E6Hm394E. |
| EMBL Accession number: | LT727359.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 02/08/2000 |
| Sequence detail: | Nucleotide sequence of the mutated E6H cDNA:
394 395
wt E6H 5'…GAG.AAC.TAC.AAG.GAC.ACC.GCA.CCA.GTC.CTA.GAC…3'
Glu Asn Tyr Lys Asp Thr Ala Pro Val Leu Asp
mutated E6H 5'…GAG.AAC.TAC.AAG.GAC.GAG.CTC.CCA.GTC.CTA.GAC…3'
Glu Asn Tyr Lys Asp Glu Leu Pro Val Leu Asp
SacI
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglI/SnaBI, ClaI/XbaI, EcoRI, SacI/XmnI, 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: |
pSV51E6HmE (LMBP 3602) 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.