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GeneCorner plasmid details

Last data update: 25 October 2020 04:11 CET

Plasmid name: pCAGGS-mCASP-7a (LMBP 4868)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4868.gb (View with Genome Compiler)
p4868.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse cysteinyl aspartate specific proteinase 7 cDNA (caspase-7, CASP-7, Casp7, Mch3)
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; pGAD-mCASP-7
Further information: The plasmid was constructed by inserting the mouse caspase-7 cDNA as a PvuII/SmaI fragment of pGAD-mCASP-7 into the MscI (BalI) opened pCAGGS vector. The orientation of the inserted fragment is antisense relative to the AG promoter.
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.
The EcoRI site at position 1183 could not be experimentally confirmed.
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 LT727050.1.
The nucleotide sequence of the mCASP-7 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number Y13088.1.
Other name of the plasmid is pCAGGS B64#5(antisense).
EMBL Accession number: Y13088.1, view at EMBL, GenBank, DDBJ
LT727050.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 23/02/2000
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, EcoRI, HindII, MspA1I, NcoI/SalI and XbaI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr M. Van de Craen(1)(2) and Prof. Dr P. Vandenabeele(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: Van de Craen et al., FEBS Lett. 403 (1997), 61-69 [PMID: 9038361]
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: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: LMBP 03819

Refer in your Materials and Methods:

pCAGGS-mCASP-7a (LMBP 4868) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Van de Craen and Prof. Dr P. Vandenabeele.

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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