Last data update: 24 January 2024 16:39 CET
Plasmid name: pRcCMVhuIKB-alpha (LMBP 3129)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3129.gb
(View with Genome Compiler) p3129.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human nuclear factor κB (NF-κB) inhibitor alpha cDNA (IκBα, NFKBIA, MAD-3) |
| Promoter: | Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer Simian virus 40 early promoter (SV40 early) Phage T7 gene 10 promoter (T7g10) Phage SP6 promoter |
| Ribosome binding site: |
- |
| Terminator: | Bovine growth hormone polyadenylation signal (BGH polyA) Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) Neomycin (neo; G418) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli Mammalian cells; SV40 permissive cells |
| Parental clone: | pRc/CMV; pCR1000 |
| Further information: | The plasmid was constructed as follows: a cDNA encoding the human IκB protein MAD-3 was amplified by the PCR technique from a human uterus cDNA library using different primers (Zabel et al. (1993)). The resulting PCR product was cloned into pCR1000. A HindIII fragment of this intermediate construct, containing the full-length human MAD-3 cDNA (hMAD3) was then inserted into the unique HindIII site of the eukaryotic expression vector pRcCMV. The plasmid is useful for expression in mammalian cells. There is uncertainty about the presence of a second enhancer in the SV40 early promoter. The absence of a second enhancer in this promoter may cause bacterial leakage expression of the Tn5 neomycin resistance gene. This would cause transformed E. coli cells to be resistant to kanamycin, although growth should be reduced compared to growth on medium containing ampicillin. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727445.1. The nucleotide sequence of the hMAD3 cDNA corresponds with the EMBL Nucleotide Sequence Database (Release 30; Accession number M69043). Other name of the plasmid is pRcCMVMAD3. |
| EMBL Accession number: | M69043, view at EMBL, GenBank, DDBJ LT727445.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 27/01/1995 |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: ApaI, BglII, HindIII and XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr P.A. Baeuerle(1) and constructed by T. Henkel(2). (1) Tularik Inc., South San Francisco, USA (2) Institute for Biochemistry, University of Freiburg, Germany |
| Plasmid reference: | Zabel et al., EMBO J. 12 (1993), 201-211 [PMID: 7679069] |
| Related plasmid reference: | Haskill et al., Cell 65 (1991), 1281-1289 [PMID: 1829648] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 XL1-Blue |
| Host reference: | Bullock et al., BioTechniques 5 (1987), 376-379 |
| 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: |
pRcCMVhuIKB-alpha (LMBP 3129) is available at BCCM/GeneCorner. This plasmid was deposited by Dr P.A. Baeuerle and constructed by T. Henkel and was published in Zabel et al., 1993. |
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.