Last data update: 22 October 2020 04:25 CEST
Plasmid name: pCAGGS-TS (LMBP 4167)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Trypanosoma cruzi trans-sialidase gene (TS); mature sequence
|Promoter:||Chicken β-actin/rabbit β-globin hybrid promoter (AG)
Human cytomegalovirus immediate early promoter (CMV-IE); enhancer only
Escherichia coli lac operon promoter
|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
|Further information:||The plasmid was constructed by inserting the Pfu amplified PCR fragment of the T. cruzi TS gene into the MscI site of pCAGGS. As a result, a unique MscI and an extra NdeI restriction site were created upstream of the start of the coding TS sequence, and a unique BclI and an extra StuI site were created downstream from the coding TS sequence.
The TS gene does not contain a C-terminal repeat domain.
pCAGGS-TS is useful for highly efficient expression of trans-sialidase under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
The sequence corresponds with the EMBL Nucleotide Sequence Database accession number AJ276679 (version SV 1).
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 LT727318.1.
Other name of the plasmid is pCAGGS-TSj.
|EMBL Accession number:||AJ276679, view at EMBL, GenBank, DDBJ
LT727318.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||25/08/2000|
Primers used to amplify the TS coding sequence: Forward: 5' GCCATGGCCAT.ATG.CTG.GCA.CCC.GGA.TCG 3' *** Reverse: 5' GGCCTAGGCCTGA.TCA.GGC.ACT.CGT.GTC.GCT.GC 3' +++ ***: start codon. +++: termination codon. Punctuation indicates reading frame
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BglII, MscI and StuI.|
|History of deposit:||This plasmid was deposited by Prof. Dr N. Callewaert(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Plasmid reference:||Laroy et al., Protein Expr. Purif. 20 (2000), 389-393 [PMID: 11087678]
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-TS (LMBP 4167) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr N. Callewaert and was published in Laroy et al., 2000.|
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.