Last data update: 24 January 2024 16:39 CET
Plasmid name: pSV71BLAL2HI (LMBP 2548)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p2548.gb
(View with Genome Compiler) p2548.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Ampicillin resistance gene (amp) Human immunoglobulin γ3 cDNA (hIG3); heavy chain hinge region |
| Promoter: | Simian virus 40 late promoter (SV40 late) Simian virus 40 early promoter (SV40 early) |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Chloramphenicol (cam) |
| Replicon: | Escherichia coli plasmid pMB1 origin Simian virus 40 bidirectional origin (SV40) |
| Host range: | Escherichia coli; preferably recombination-deficient strains Mammalian cells; SV40 permissive cells |
| Parental clone: | pSV71BLAHI; pMacBLAL2HI |
| Further information: | The plasmid was constructed by substituting the SacI-AhaII fragment of pSV71BLAHI by the SacI-AhaII fragment of pMacBLAL2HI. This plasmid leads to the expression of a fusion protein in which 10 additional amino acids (derived from the VH region of OMMB) are present between β-lactamase and the first hinge region (H1) of the human γ3 heavy chain. The translation of this fusion protein terminates 8 amino acids behind the fourth hinge region (H4). It differs in this respect from the BlAL2Hi fusion protein expressed from pMacBLAL2HI. The bacterial promoter in front of the ampicillin resistance gene is absent, so ampicillin can't be used as a selection marker; after expression in COS-1 cells, however, β-lactamase is enzymatically active. This plasmid contains a repeated sequence; hence, watch out for homologous recombination. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727527.1. |
| EMBL Accession number: | LT727527.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 22/01/1997 |
| Sequence detail: | Nucleotide sequence at the fusion between amp and the hinge region of hIG3:
amp linker (1) H1-4
------- -------------------- ---------------
5' CAT TGG GAG TCG GGC CCA .... GAG CTC ... CCA GGG CCC ... TAG 3'
SacI ^ ApaI *
^: End of the fourth hinge region (H4).
*: Termination codon (8 amino acids behind H4).
(1): The 10 amino acid linker segment, derived from the C-terminal end of VH. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: EcoRI/PstI, EcoRV, NcoI/PvuI, SacI/XmnI, StyI and XbaI/XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr K. De Sutter(1). (1) 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) + chloramphenicol (25 μg/ml)* |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | *: selection of transformants on chloramphenicol; subsequent cultivation of a single colony in liquid medium with ampicillin. |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSV71BLAL2HI (LMBP 2548) is available at BCCM/GeneCorner. This plasmid was deposited by Dr K. De Sutter. |
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.