Last data update: 24 January 2024 16:39 CET
Plasmid name: pCAGGS-E-hA20-3R/3A (LMBP 5936)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | not available |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human TNF alpha induced protein 3 cDNA (TNFAIP3, A20, OTUD7C, GeneID 7128); mutated sequence E-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: | pCAGGSEhA20 |
| Further information: | The plasmid was constructed by inserting a HpaI-SacI PCR fragment, containing the mutated sequence (three arginine residues were mutated to alanine) of hA20, between the HpaI and SacI site of pCAGGSEhA20. Human A20 is mutated in three possible cleavage sites for MALT1. pCAGGS-E-hA20-3R/3A is useful for highly efficient expression of hA20-3R/3A under the control of the AG promoter and the hCMV-IE enhancer in various mammalian cells. 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. |
| EMBL Accession number: | M59465, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 03/02/2009 |
| Sequence detail: | PCR mutagenesis on pCAGGSEhA20 was conducted with the following primers:
271 275 279
F1-hA20-HpaI: 5' C.CGA.GCT.GTT.CCA.CTT.GTT.AAC.AGA.GAC 3'
HpaI
443 440 439 435 434
R3-hA20-RA: 5' ATA.GGC.TTC.TCC.CGC.AGA.GGC.CCC.GAG.CGC.C 3'
^^
406 410 411 415 418
F2-hA20-RRAA: 5' GAG.TGC.TCA.GAG.GCG.GCG.CAA.AAG.AAT.CAA.AAC.AAA.CTC 3'
Ala Ala
^^ ^^
415 411 410 405 403
R2-hA20-RRAA: 5' TTG.ATT.CTT.TTG.CGC.CGC.CTC.TGA.GCA.CTC.ATG.GCA.TAA.AG 3'
^^ ^^
436 439 440 445 446
F3-hA20-RA: 5' TC.GGG.GCC.TCT.GCG.GGA.GAA.GCC.TAT.GAG.CCC.TTG 3'
Ala
^^
737 735 730
R1-hA20-SacI: 3' A.TGC.TGA.CAC.TCC.ATG.CAG.AGC.TCC 3'
SacI
410 411 439
wt hA20: 5' ... GAG.AGG.CGG.CAA ... TCT.CGG.GGA
Arg Arg Arg
^: mutated nucleotide.
Punctuation indicates reading frame. |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| 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-E-hA20-3R/3A (LMBP 5936) 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.