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GeneCorner plasmid details

Last data update: 23 September 2021 14:51 CEST

Plasmid name: pGBF-Kozakcre (LMBP 4474)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: p4474.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Phage P1 cre gene encoding recombinase; with Kozak consensus sequence
Promoter: Escherichia coli lac operon promoter
Phage T7 gene 10 promoter (T7g10)
Phage T3 promoter
Ribosome
binding site:
-
Terminator: -
Selection marker: Ampicillin (amp)
Neomycin (neo; kanamycin (kan))
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli
Parental clone: pCR4TOPO
Further information: The plasmid was constructed by PCR amplifying the cre coding sequence from phage P1 and cloning it into pCR4TOPO. The orientation of the inserted fragment is sense relative to the T7 promoter.
The ATG initiation codon is embedded in a consensus Kozak sequence to optimize translation initiation in mammalian cells.
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 the cre gene corresponds with the EMBL Nucleotide Sequence Database accession number X03453.1.
EMBL Accession number: X03453.1, view at EMBL, GenBank, DDBJ
Latest sequence update: 17/12/2001
Sequence detail:
forward primer: 5' GCTAGCCCACC.ATG.TCC.AAT.TTA.CTG.ACC.GTA.C 3'
                   NheI        ***

reverse primer: 5' ACTAGT.CTA.ATC.GCC.ATC.TTC.CAG.CAG 3'
                   SpeI   +++

***: start codon.
+++: termination codon.
Punctuation indicates reading frame.
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: BamHI, BglI/XmnI, HaeII and NheI/SpeI.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Prof. Dr W. Muller(1).
(1) Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom
Plasmid reference: -
Related plasmid reference: Sternberg et al., J. Mol. Biol.187 (1986), 197-212 [PMID: 3486297]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 TOP10
Host reference: Invitrogen Instruction Manual
Related host reference: Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051]
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:

pGBF-Kozakcre (LMBP 4474) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr W. Muller.

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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