GREAT AT SMALL THINGS

0

GeneCorner plasmid details

Last data update: 29 October 2020 04:15 CET

Plasmid name: pBMGNeoE6Hy3f2 (LMBP 1974)

Add to cart

New search

Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p1974.gb
Sequence
analysis results
Genecorner:

-

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)
Ribosome
binding site:
-
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
Mammalian cells
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
Sequence detail:
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.
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: -
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)
Cultivation temperature: 37°C
Biosafety level: L1
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.

New search