Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGSFLAGmCASP-11m2 (LMBP 3972)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p3972.gb
(View with Genome Compiler) p3972.txt p3972.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Mouse cysteinyl aspartate specific proteinase 11 cDNA (caspase-11, CASP-11, Ich3, Casp4); mutated sequence FLAG epitope tag; N-terminal |
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-FLAGmA20f3 |
Further information: | The plasmid was constructed by inserting a KpnI/SalI PCR amplicon, encoding mutated mCASP-11, in the KpnI/SalI opened pCAGGS-FLAGmA20f3 vector. The mCASP-11 insert contains a Cys/Ser mutation at codon position 254, creating a unique NheI site. pCAGGSFLAGmCASP-11m2 is useful for highly efficient expression of mutated mCASP-11 under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells. 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 LT727297.1. The nucleotide sequence of the mCASP-11 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number Y13089.1, except for the 3' end of the coding sequence that corresponds with the nucleotide sequence of the reverse primer. Other names of the plasmid are pCAGGSFLAGCASP-11M and pCAGGSFLAGCASP-11MCys/Ser. |
EMBL Accession number: | Y13089.1, view at EMBL, GenBank, DDBJ LT727297.1, view at EMBL, GenBank, DDBJ |
Latest sequence update: | 07/09/2007 |
Sequence detail: | Primers used to amplify mutated mCASP-11: Forward primer: 5' CCATGGGTACC.ATG.GCT.GAA.AAC.AAA.CAC.CCT.GAC 3' KpnI *** Reverse primer: 5' ACGCGTCGAC.TCA.GTT.GCC.AGG.AAA.GAC.GTA.GAA.ATA 3' SalI +++ | G according to EMBL Y13089 (C results in Val, G in Leu) ***: start codon. +++: termination codon. Punctuation indicates reading frame. Nucleotide sequence of the mutated mCASP-11 cDNA: 250 253 254 wild type: 5' ATT.GTG.CAG.GCC.TGC.AGA.GGT.GGG.AAC 3' Ile Val Gln Ala Cys Arg Gly Gly Asn 250 253 254 mutated: 5' ATT.GTG.CAG.GCT.AGC.AGA.GGT.GGG.AAC 3' Ile Val Gln Ala Ser Arg Gly Gly Asn NheI ^ ^ ^: mutated nucleotide. Punctuation indicates reading frame. |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglII, EcoRI, NcoI, NdeI, XbaI and XhoI. |
Class: | Recombinant plasmid |
Type: | Plasmid |
History of deposit: | This plasmid was deposited by Prof. Dr R. Beyaert(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: | Schoen et al., PLoS One. 8 (2013), e56520 [PMID: 23441200] [DOI: 10.1371/journal.pone.0056520] Wang et al., Cell 92 (1998), 501-509 [PMID: 9491891] |
Restricted distribution: | - BCCM MTA |
Distributed as: | H/P active culture or plasmid DNA |
Host for distribution: | Escherichia coli K12 MC1061 |
Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
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: |
pCAGGSFLAGmCASP-11m2 (LMBP 3972) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert . |
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.