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GeneCorner plasmid details

Last data update: 22 October 2020 04:25 CEST

Plasmid name: pCAGGS-TS (LMBP 4167)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p4167.gb (View with Genome Compiler)
p4167.txt
p4167.pdf
Sequence
analysis results
Genecorner:

-

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
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
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
Sequence detail:
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.
Class: Recombinant plasmid
Type: Plasmid
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)
Cultivation temperature: 37°C
Biosafety level: L1
Cultivation remark: -
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.

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