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GeneCorner plasmid details

Last data update: 26 November 2020 04:30 CET

Plasmid name: pSV25SE6Hf2 (LMBP 1703)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1703.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mouse anti-hPLAP monoclonal antibody E6(IgG2b,κ) cDNA; truncated heavy chain of the F(ab')2 fragment E6F2 (E6Hf2)
Promoter: 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: pSV25S; pSV23SE6H
Further information: E6Hf2 is a HindIII fragment of E6H (the cDNA copy of the heavy chain of the mouse monoclonal antobody E6), isolated from pSV23SE6H); the second HindIII site is made by filling in the unique XhoI site of pSV23SE6H (situated in the first part of CH2), and adding at least 2 multistoplinkers : 'CTTAGTTAACTAAG' (MSL); the succession of the 2 linkers resulted in the formation of this second HindIII site.
The multi-stop-linker contains a HpaI site.
The E6Hf2 gene is under control of the early promoter of SV40 and is removable as a HindIII fragment.
This plasmid allows the synthesis of a truncated mouse heavy chain of the F(ab')2 fragment E6F2 wich contains a CH2 domain of only 8 amino acids.
The residual fragment of the CH2 domain (nucl. 1659-1633) is too small to be displayed on the circular map.
Designed for transient expression in COS cells or constitutive expression in CHO cells (cotransfection with DHFR plasmids).
Name of the plasmid mentioned in Feys et al. (1988) is pSV25pE6F2.
EMBL Accession number: -
Latest sequence update: 28/03/1989
Sequence detail:
Nucleotide sequende of the CH2 domain fragment:

   Lys Cys Pro Ala Pro Asn Leu Asp Leu Val Asn *
5' AAA TGC CCA GCT CCT AAC CTC GAC TTA GTT AAC TAA GCTTGGC 3'
           ^   +           Filled-in   HpaI     HindIII
                           XhoI
                                 ___________________
                                       linker

^: End hinge.
+: Start CH2 fragment.
*: Termination codon.
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 V. Feys(1) and Prof. Dr W. Fiers(1).
(1) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Feys et al., Int. J. Cancer Suppl. 2 (1988), 26-27 [PMID: 3162443]
Related plasmid reference: De Sutter et al., Gene 113 (1992), 223-230 [PMID: 1572543]
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:

pSV25SE6Hf2 (LMBP 1703) is available at BCCM/GeneCorner. This plasmid was deposited by V. Feys and Prof. Dr W. Fiers and was published in Feys et al., 1988.

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.

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