Last data update: 23 October 2020 04:27 CEST
Plasmid name: pCAGGS-TAX1BP1-L (LMBP 4050)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human T-cell leukemia virus type I (HTLV-1) Tax1 binding protein 1 cDNA (TXBP15-1; TXBP151; TAX1BP1); large form (TAX1BP1-L)
|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
|Parental clone:||pCAGGS2; pCAGGS-TAX1BP1-S; pCAGGSTXBP151-L|
|Further information:||The plasmid was constructed by inserting a 1488 bp XbaI-BglII fragment from pCAGGSTXBP151-L and a 967 bp BglII-XhoI fragment from pCAGGS-TAX1BP1-S, into the XbaI/XhoI opened pCAGGS2 vector.
pCAGGS-TAX1BP1-L is useful for highly efficient expression of the large human Tax1 binding protein 1 under the control of the AG promoter and the human CMV-IE enhancer in various mammalian cells.
Since the sequence around the EcoRI site at the 5' end of the TAX1 insert could not be completely traced back, there is uncertainty as to the cutting frequency of the restriction enzymes, except for the sites that were analysed in the authenticity test.
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 LT727299.1.
The TAX1BP1-L cDNA was sequenced at the Department of Biomedical Molecular Biology (Ghent University, Belgium). Referring to the sequence analysis results, the human TAX1BP1-L coding sequence corresponds with the EMBL Nucleotide Sequence Database accession number U33821.2 except for: 1) nucleotides CG at position 699-700 are substituted by GC, resulting in the replacement of amino acid 233 (His -> Gln) and 234 (Val -> Leu); 2) nucleotide T at position 919 is substituted by A, resulting in a Leu -> Ile change of amino acid 307; 3) nucleotide T at position 1755 is substituted by C, whereby amino acid 585 remains Asp. These differences were confirmed by Dr K-T. Jeang (NIH, Bethesda, Maryland, USA) who deposited pSV/TXBP15-1 (LMBP 3514) and pSV/TXBP15-1-AS (LMBP 3515).
Other name of the plasmid is pCAGGS-TAX1BP1.
|EMBL Accession number:||U33821.2, view at EMBL, GenBank, DDBJ
LT727299.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||31/01/2002|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: BglII, NcoI, XbaI and XhoI.|
|History of deposit:||This plasmid was deposited by Prof. Dr R. Beyaert(1) (2). It was constructed by M. Klinkenberg(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
|Related plasmid reference:||De Valck et al., Oncogene 18 (1999), 4182 - 4190 [PMID: 10435631]
Shembade et al., EMBO J. 26 (2007), 3910-3922 [PMID: 17703191] [DOI: 10.1038/sj.emboj.7601823]
|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)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pCAGGS-TAX1BP1-L (LMBP 4050) 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.