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GeneCorner plasmid details

Last data update: 24 January 2024 16:39 CET

Plasmid name: pSEAPBhBCTNf3 (LMBP 3783)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p3783.gb (View with Genome Compiler)
p3783.txt
p3783.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human placental alkaline phosphatase gene (PLAP, ALPP); mutant encoding a secreted protein (SEAP)
Promoter: Human β-catenin promoter (BCTN, CTNNB1)
Ribosome
binding site:
-
Terminator: Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Host range: Escherichia coli
Mammalian cells
Parental clone: pSEAPBasic
Further information: The plasmid was constructed by inserting the 5,6 kb EcoRI fragment (filled in with T4 DNA polymerase) of the human CTNNB1 gene, containing 3,4 kb of the 5' flanking region, the CTNNB1 promoter sequence, the non-coding exon 1 (5' untranslated region) and 1 kb of intron 1, into the unique, T4 DNA polymerase filled-in HindIII site of pSEAPBasic.
The SV40 small t-antigen splice signal and the SV40 polyA's downstream from the SEAP gene ensure efficient processing.
The SV40 polyA upstream of the 5' flanking region of the human CTNNB1 gene reduces background transcription.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727388.1.
The nucleotide sequence of pSEAPBasic corresponds with the EMBL Nucleotide Sequence Database accession number U09660.1
The nucleotide sequence of the human CTNNB1 promoter corresponds with the EMBL Nucleotide Sequence Database accession number X89448.1
The nucleotide sequence of the 5' flanking region, the 5' untranslated region and the first intron of the human CTNNB1 gene corresponds with the Genbank accession number NT_022517.18.
Name mentioned in Nollet et al. (1996) is pSEAP Basic / FRAG3.
Other name of the plasmid is pSEAPBhBCTN-FRAG3.
EMBL Accession number: X89448.1, view at EMBL, GenBank, DDBJ
U09660.1, view at EMBL, GenBank, DDBJ
NT_022517.18, view at EMBL, GenBank, DDBJ
LT727388.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 16/09/2010
Sequence detail:
Nucleotide sequence around the termination codon of the human PLAP gene:

                485 486 487 488 489 490 491 492 493      513
PLAP:     5'... GCC GCG CAC CCG GGG CGG TCC GTG GTC......CCC TGA ... 3'
                Ala Ala His Pro Gly Arg Ser Val Val      Pro +++

SEAP:     5'... GCC GCG CAC CCG GGT TAA CCCGTGGTCCCC ... 3'
                Ala Ala His Pro Gly +++
                                 HpaI
+++: termination codon.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglII, EcoNI, SfiI/SphI, XcmI and XhoI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr F. Nollet(1) (2) and Prof. Dr F. Van Roy(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Nollet et al., Genomics 32 (1996), 413-424 [PMID: 8838805]
Related plasmid reference: Previdi et al., Eur. J. Cancer 46 (2010), 1679-1691 [PMID: 20350802] [DOI: 10.1016/j.ejca.2010.02.036]
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 + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: -

Refer in your Materials and Methods:

pSEAPBhBCTNf3 (LMBP 3783) is available at BCCM/GeneCorner. This plasmid was deposited by Dr F. Nollet and Prof. Dr F. Van Roy and was published in Nollet et al., 1996.

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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