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GeneCorner plasmid details

Last data update: 23 November 2020 04:19 CET

Plasmid name: pCAGGS-FLAG-TAX1BP1-L2 (LMBP 5665)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner core plasmid
GeneCorner sequence: p5665.gb (View with Genome Compiler)
p5665.txt
p5665.pdf
Sequence
analysis results
Genecorner:

-

Cloned DNA: Human T-cell leukemia virus type I (HTLV-1) Tax1 binding protein 1 cDNA (TXBP15-1; TXBP151; TAX1BP1); large form (TAX1BP1-L)
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: pcDNA3flag-Tax1BP1; pCAGGS-FLAG-TAX1BP1-L
Further information: The plasmid was constructed by inserting a BglII PCR fragment, amplified from pcDNA3flag-Tax1BP1 and containing the C-terminal part of the human TAX1BP1 coding sequence, into the BglII opened pCAGGS-FLAG-TAX1BP1-L.
The plasmid is useful for highly efficient expression of human TAX1BP1-L2 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 lacIq 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 LT726939.1.
According to the depositor, the human TAX1BP1-L2 CDS is a slightly longer splice variant compared to the human TAX1BP1-L CDS of the plasmid pCAGGS-FLAG-TAX1BP1-L (LMBP 5163). According to sequencing results however, the only difference with pCAGGS-FLAG-TAX1BP1-L is that pCAGGS-FLAG-TAX1BP1-L2 lacks a fragment of pCAGGS upstream of the 3' UTR of the rabbit β-globin gene. The depositor will be contacted to obtain the longer splice variant.
The entire human TAX1BP1 CDS was sequenced at LMBP.
The nucleotide sequence of the human TAX1BP1 coding sequence corresponds with the Genbank accession number BC001764.1, except for an A instead of T at nucleotide position 919 of the human TAX1BP1 CDS, resulting in a isoleucine instead of leucine at amino acid position 307. As compared to the EMBL accession number U33821.2, there are more nucleotide differences.
EMBL Accession number: BC001764.1, view at EMBL, GenBank, DDBJ
LT726939.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 09/10/2013
Sequence detail:
Primers used to amplify the C-terminal part of human TAX1BP1:

forward: 5' GAA.GAT.CTG.AAA.CTC.CGT.CTT.C 3' 
              BglII

reverse: 5' GAAGATCT.CTA.GTC.AAA.ATT.TAG.AAC.ATT.C 3'
              BglII  +++

+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BglII/XhoI, EcoRI, SpeI and XbaI/XmnI.
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: -
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-FLAG-TAX1BP1-L2 (LMBP 5665) 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.

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