Last data update: 24 January 2024 16:39 CET
Plasmid name: pAS2A20ZF- (LMBP 3767)
New search | Print data sheet |
Price category: | Cat. 1 (cf. price list) |
Status: | GeneCorner core plasmid |
GeneCorner sequence: |
p3767.gb
(View with Genome Compiler) p3767.pdf |
Sequence analysis results Genecorner: |
- |
Cloned DNA: |
Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128); with C-terminal deletion Saccharomyces cerevisiae GAL4 DNA-binding domain (GALbd) Influenza HA epitope encoding the haemagglutinin tagging peptide; N-terminal |
Promoter: | Saccharomyces cerevisiae alcohol dehydrogenase promoter (ADH1) Escherichia coli lac operon promoter |
Ribosome binding site: |
- |
Terminator: | Saccharomyces cerevisiae alcohol dehydrogenase terminator (ADH1) Saccharomyces cerevisiae iso-1-cytochrome C terminator (CYC1) Saccharomyces cerevisiae 2 micron plasmid (2μ) FLP terminator |
Selection marker: | Ampicillin (amp) Cycloheximide (CYH2) TRP1; auxotrophic |
Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Saccharomyces cerevisiae 2 micron plasmid origin (2μ); incl. region conferring stability (STB) |
Host range: | Escherichia coli Saccharomyces cerevisiae; trp1(-) |
Parental clone: | pAS2; pLT10T3A20 |
Further information: | The plasmid was constructed by inserting the 1101 bp NdeI-Bsp1286I (blunted with T4 DNA polymerase) fragment of pLT10T3A20, containing C-terminal-deleted human zinc finger protein A20 cDNA (hA20ZF(-)), between the NdeI and SmaI sites of pAS2. hA20ZF(-) spans amino acid 1 to 367 and lacks the seven Cys2/Cys2 zinc fingers. The S. cerevisiae GALbd domain is composed of the first 147 codons from GAL4, containing the nuclear localization signal. The GALbd-HA-hA20ZF(-) fusion protein can be used as a bait protein in two-hybrid screening. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727377.1. The nucleotide sequence of the pAS2 vector corresponds with the EMBL Nucleotide Sequence Database accession number U30496.1, but adapted as follows: 1) the 'N' nucleotide was removed, restoring the coding sequence of cycloheximide; 2) the sequence GATTTC between GALbd and the HA epitope was replaced by GAATTC, an EcoRI site that has been experimentally confirmed in pAS2; 3) the sequence TTCGAA (Csp45I site) in the ADH1 promoter was replaced by TCGAAG, eliminating the Csp45I site that could not be experimentally confirmed in pAS2. The nucleotide sequence of the hA20 coding sequence corresponds with the Genbank accession number M59465.1. Name mentioned in De Valck et al. (1996) is pAS2A20ZF(-). Other name of the plasmid is pAS2HuA20ZF(-). |
EMBL Accession number: | M59465.1, view at EMBL, GenBank, DDBJ LT727377.1, view at EMBL, GenBank, DDBJ |
Latest sequence update: | 03/02/2004 |
Sequence detail: | The nucleotide sequence of the fusion gene: ------------------ GALbd -----------------> 5' ... CTGAAAG.ATG.AAG.CTA.CTG.TCT ... CAG.TTG.ACT.GTA.TCG.CCG.GAA.TTC.ATG.GCT. Met Lys Leu Leu Ser Gln Leu Thr Val Ser Pro Glu Phe Met Ala *** EcoRI --------------- ----------- HA epitope -----------> 1 TAC.CCA.TAC.GAT.GTT.CCA.GAT.TAC.GCT.AGC.TTG.GGT.GGT.CAT.ATG.GCT.GAA.CAA Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Gly Gly His Met Ala Glu Gln NheI NdeI -- hA20f -----------------> 367 GTC ... GGG.AGG.AGA.GAG.GGG.GGA.TCC.GTC.GAC.CTG.CAG.CCA.AGC.TAA.TTCCGG ... 3' Val Gly Arg Arg Glu Gly Gly Ser Val Asp Leu Gln Pro Ser +++ BamHI SalI PstI HindII ***: start codon. +++: termination codon. Punctuation indicates reading frame. |
Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BamHI, HindIII and SphI. |
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: | De Valck et al., FEBS Lett. 384 (1996), 61-64 [PMID: 8797804] |
Related plasmid reference: | De Valck et al., Oncogene 18 (1999), 4182 - 4190 [PMID: 10435631] De Valck et al., Biochem. Biophys. Res. Commun. 238 (1997), 590-594 [PMID: 9299557] |
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 |
Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pAS2A20ZF- (LMBP 3767) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr R. Beyaert and was published in De Valck et al., 1996. |
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.