Last data update: 24 January 2024 16:39 CET
Plasmid name: pSEAPBhBCTNf2 (LMBP 3784)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3784.gb
(View with Genome Compiler) p3784.txt p3784.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; pSEAPBhBCTNf3; pGEM-11Zf(+) |
| Further information: | The plasmid was constructed as follows: 1) the 437 bp SacII fragment, containing 298 bp of the human CTNNB1 promoter sequence and 139 bp of the non-coding exon 1 (5' untranslated region) of the human CTNNB1 gene, was excised from pSEAPBhBCTNf3, blunted with T4 DNA polymerase and inserted into the unique HindII site of pGEM-11Zf(+); 2) the EcoRI/XhoI fragment of this intermediate construct was then filled in with T4 DNA polymerase and inserted into the Pfu 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 human CTNNB1 promoter reduces background transcription. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727389.1. 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' untranslated region of the human CTNNB1 gene corresponds with the Genbank accession number NT_022517.18. Name mentioned in Nollet et al. (1996) is pSEAP Basic / FRAG2. Other name of the plasmid is pSEAPBhBCTN-FRAG2. |
| 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 LT727389.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 17/07/2008 |
| 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: BamHI, NarI and PstI. |
| 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] |
| 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: |
pSEAPBhBCTNf2 (LMBP 3784) 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.