Last data update: 29 October 2020 04:15 CET
Plasmid name: pBMGNeoE6Hy3f2 (LMBP 1974)
|Price category:||Cat. 1 (cf. price list)|
|Status:||GeneCorner non-core plasmid|
|Cloned DNA:||Mouse anti-hPLAP monoclonal antibody E6(Hy3,κ) half genomic gene; truncated mouse::human chimeric heavy chain of the F(ab')2 fragment E6(Hy3,κ)F2 (E6Hy3f2)
|Promoter:||Mouse metallothionein 1 promoter (Mt1)
Herpes simplex virus (HSV) thymidine kinase promoter (TK)
|Terminator:||Herpes simplex virus (HSV) thymidine kinase polyadenylation signal (TK polyA)|
|Selection marker:||Ampicillin (amp)
Neomycin (neo; G418; kanamycin (kan))
|Replicon:||Escherichia coli plasmid pMB1 origin
Bovine papilloma virus origin (BPV)
|Host range:||Escherichia coli
|Parental clone:||pBMGNeo; pSV25E6Hy3f2|
|Further information:||This plasmid contains the E6Hy3f2 gene in a sense orientation relative to the mMt1 promoter.
This plasmid was made as follows: 1) The E6Hy3f2 gene was isolated as a HindIII fragment from pSV25E6Hy3f2; the 5' sticky ends were filled in with Klenow and SalI linkers ('CGTCGACG') were added; 2) This SalI fragment, containing the E6Hy3f2 gene, was then cloned into the unique SalI site of pBMGNeo, clockwise behind the mMt1 promoter.
The mMt1 promoter controls the expression of the inserted E6Hy3f2 gene in mammalian cells.
Because of the presence of the second intron of the rabbit β-globin gene, the E6Hy3f2 transcript contains an additional intron in the 3' untranslated region.
The truncated CH2 domain consists of 31 amino acids, terminating at the multi-stop-linker.
See also pBMGNeo and pSV25E6Hy3f2 for other remarks.
|EMBL Accession number:||-|
|Latest sequence update:||17/06/1991|
Terminal sequence of the truncated CH2 domain: CH2 pBMGNeo -------------- ----------> Arg Thr Pro Asp Leu Val Asn * 5' CGG ACC CCT GAC TTA GTT AAC TAA GCT CGTC GACNNNNNNN 3' ^ HpaI Filled- SalI in HindIII ------------------- linker *: Termination codon. ^: 27th amino acid of CH2.
|Authenticity test:||The plasmid still needs to be subjected to the authenticity test.|
|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
|Restricted distribution:||- BCCM MTA|
|Distributed as:||H/P active culture or plasmid DNA|
|Host for distribution:||Escherichia coli K12 DH5α|
|Host reference:||Focus 8 (1986), 9
|Related host reference:||Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187]
Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791]
Rodriguez-Quinones et al., Focus 15 (1993), 110-112
Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660]
|Cultivation medium:||LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml)|
|Other culture collection numbers:||-|
Refer in your Materials and Methods:
|pBMGNeoE6Hy3f2 (LMBP 1974) is available at BCCM/GeneCorner. This plasmid was deposited by V. Feys and Prof. Dr W. Fiers.|
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.