Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV-Sport-PITSLRE-E (LMBP 5267)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p5267.gb
(View with Genome Compiler) p5267.txt p5267.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human PITSLRE kinase cDNA (CDC2L2, CDK11, p110 PITSLRE); expressing the β SV3 isoform E-tag; C-terminal |
| Promoter: | Phage SP6 promoter Phage T7 gene 10 promoter (T7g10) Escherichia coli lac operon promoter; mutant (lacUV5) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | 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: | pSV-Sport-Etag; pMa-p110PITSLRE |
| Further information: | This plasmid was constructed as follows: 1) CDC2L2 cDNA was amplified by PCR; 2) the amplicon was EcoRI digested and cloned into pMa58; 3) a NotI restriction site was introduced at the CDC2L2 termination codon by site-directed mutagenesis; 4) the CDC2L2 cDNA was finally inserted as an EcoRI-NotI fragment in the pSV-Sport-Etag expression vector, in frame with the C-terminal E-tag. 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. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT726995.1. The nucleotide sequence of the CDC2L2 cDNA corresponds with the EMBL Nucleotide Sequence Database accession number AF067521.1, adapted to the BCCM/GeneCorner results of the sequence analysis of the 5' side of the insert. The following difference is observable: the nucleotide T at position 892 of the CDC2L2 coding sequence is substituted by a C-residue, resulting in a Ser/Pro replacement for amino acid 298. The sequencing results of the 3' side of the insert revealed several differences with AF067521, including the absence of the EcoRI site in the CDC2L2 cDNA. The plasmid sequence file was not adapted to these observations. |
| EMBL Accession number: | AF067521.1, view at EMBL, GenBank, DDBJ LT726995.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 23/05/2008 |
| Sequence detail: | Sequence at the borders of the CDC2L2 coding sequence:
|-> CDC2L2 ->|
5' ... GAATTCTTAACTCAA|ATG.GGT.GAT.GAA.AAG.GAC.TCT.TGG ... CTC.AAG.TTC|
EcoRI
|-> E-tag ->|
GCG.GCC|GCA.GGT.GCG.CCG.GTG.CCG.TAT.CCG.GAT.CCG.CTG.GAA.CCG.CGT|GCC.GCA.TAG ... 3'
NotI ***
***: termination codon. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI/NotI, HaeII and XbaI. The EcoRI site in the CDC2L2 cDNA could not be experimentally confirmed. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Y. Bruynooghe(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 |
| Plasmid reference: | Cornelis et al., Mol. Cell 5 (2000), 597-605 [PMID: 10882096] |
| 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: |
pSV-Sport-PITSLRE-E (LMBP 5267) is available at BCCM/GeneCorner. This plasmid was deposited by Y. Bruynooghe and Prof. Dr R. Beyaert and was published in Cornelis et al., 2000. |
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.