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GeneCorner plasmid details

Last data update: 21 October 2020 04:26 CEST

Plasmid name: pCAGGSE6L (LMBP 3547)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3547.gb (View with Genome Compiler)
p3547.pdf
Sequence
analysis results
Genecorner:

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Cloned DNA: Mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) cDNA; light chain (E6L)
Promoter: Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
Ribosome
binding site:
-
Terminator: Rabbit β-globin polyadenylation signal (β-globin polyA)
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: pCAGGS; pSV51E6L1
Further information: The plasmid was constructed by ligating the XbaI fragment (filled in with Klenow DNA polymerase) from pSV51E6L1, containing a cDNA copy of the light chain of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ), to the MscI vector fragment from pCAGGS.
pCAGGSE6L is useful for efficient expression of the light chain of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) under the control of the chicken β-actin/rabbit β-globin hybrid promoter (AG) and the human CMV-IE enhancer in various mammalian cells.
In combination with an E6H (heavy chain of the mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ)) expression vector (e.g. pCAGGSSE6H), this plasmid can be used to produce a hPLAP (human placental alkaline phosphatase)-specific mouse antibody.
In combination with an expression vector for a heavy chain mouse immunotoxin (e.g. pUHDE6Hf2ETA) or a mouse::human immunotoxin (e.g. pUHDE6Hy3f2ANG), this plasmid can be used to produce a hPLAP-specific mouse or mouse::human chimeric immunotoxin respectively.
The AG promoter sequence consists of the chicken β-actin promoter, the first exon and part of the first intron (that seems to have a strong enhancer-like activity) linked to a rabbit β-globin fragment, consisting of a 3' part of the second intron (inclusive a branch point which is required for normal splicing reactions) and a 5' part of the third exon.
When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727350.1.
EMBL Accession number: LT727350.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 31/08/2000
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, BamHI/XmnI, BglII/XhoI, SacII, SacII/SspI, SspI and XmnI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr R. Contreras(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: -
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061(λ)
Host reference: Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329]
Related host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pCAGGSE6L (LMBP 3547) is available at BCCM/GeneCorner. This plasmid was deposited by 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.

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