Last data update: 23 October 2020 04:27 CEST
Plasmid name: pCAGGS-FLAG-STING-SV (LMBP 6922)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner core plasmid|
(View with Genome Compiler)
|Cloned DNA:||Human transmembrane protein 173 (TMEM173, NET23, ERIS, MITA, STING); splice variant
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
|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 ligating the PCR amplified human TMEM173 splice variant coding sequence as an EcoRI/SalI fragment into the EcoRI/Sal opened pCAGGS-FLAG-DUBA vector.
The endoplasmic reticulum transmembrane protein TMEM173 forms dimers upon activation, leading to Interferon Regulatory Factor 3 (IRF3) activation and subsequent induction of interferon (IFN).
The FLAG epitope tag contains an enterokinase cleavage site to allow removal.
The splice variant lacks basepairs 760 to 946, resulting in a frameshift and a truncated protein of approximately 31 kDa.
The depositor confirmed mammalian expression by Western blotting.
The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726846.1.
The nucleotide sequence of the human TMEM173 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number BC047779.1, except for the following nucleotide changes in the coding sequence which were experimentally determined by the depositor: A instead of G in position 212, C instead of G in position 689 and G instead of A in position 695. The nucleotide changes in positions 212 and 695 are known 'single nucleotide polymorphisms (SNPs)', leading to R71H and H232R amino acid changes. The nucleotide change in position 689 of the coding sequence leads to a G230A amino acid change.
Other name of the plasmid is pCAGGS-flag-Sting SV.
|EMBL Accession number:||BC047779.1, view at EMBL, GenBank, DDBJ
LT726846.1, view at EMBL, GenBank, DDBJ
|Latest sequence update:||19/07/2013|
|Authenticity test:||Restriction enzyme pattern analysed at GeneCorner: ApaI/HindIII, EcoRI/SalI, NcoI, SacI and ScaI.|
|History of deposit:||This plasmid was deposited by Dr S. Gardam(1) (2) and 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
|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-FLAG-STING-SV (LMBP 6922) is available at BCCM/GeneCorner. This plasmid was deposited by Dr S. Gardam and 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.