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LMBP plasmid details

Last data update: 26 September 2020 04:22 CEST

Plasmid name: pLenti6-hRIPK4-K51R-VSV-puro (LMBP 10186)

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

-

Cloned DNA: Human receptor-interacting serine-threonine kinase 4 cDNA (RIPK4, RIP4, ANKRD3, ANKK2, DIK, PKK); mutated sequence
Vesicular stomatitis virus glycoprotein (VSV-G) epitope; C-terminal
Promoter: Human cytomegalovirus immediate early promoter (CMV-IE) and enhancer
Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 5' LTR)
Simian virus 40 early promoter (SV40 early)
Escherichia coli lac operon promoter
Phage T3 promoter
Phage T7 gene 10 promoter (T7g10)
Ribosome
binding site:
-
Terminator: Rous sarcoma virus/human immunodeficiency virus hybrid long terminal repeat (RSV/HIV-1 3' LTR); modified HIV-1 3' LTR with deleted U3 region (ΔU3/3' LTR)
Simian virus 40 polyadenylation signal (SV40 polyA)
Selection marker: Ampicillin (amp)
Puromycin (puro)
Replicon: Escherichia coli plasmid pMB1 origin
Phage f1 origin
Simian virus 40 bidirectional origin (SV40)
Host range: Escherichia coli
Mammalian cells; SV40 permissive cells
Parental clone: pLenti6-hRIPK4-VSV-puro
Further information: This Gateway expression plasmid was constructed by introducing a K51R mutation into the human RIPK4 coding sequence of pLenti6-hRIPK4-VSV-puro via site-directed mutagenesis.
Two more nucleotides directly upstream of the K51R mutation have been changed, resulting in silent mutations.
The region from the VSV epitope up to and including the puromycin resistance gene has been sequenced at GeneCorner.
Because the risk for recombination after each subcultivation is high, this plasmid is only available under the format of isolated plasmid DNA.
Other name of the plasmid is pLenti6puroVSV_hRIPK4 K51R.
EMBL Accession number: -
Latest sequence update: 01/12/2016
Sequence detail:
Wild type sequence of the human RIPK4 coding sequence:

   48  49  50  51  52  53
...CTG.GCC.ATC.AAG.TGC.TCG...
   Leu Ala Ile Lys Cys Ser

Mutated sequence of the human RIPK4 coding sequence:

   48  49  50  51  52  53
...CTC.GCG.ATC.AGG.TGC.TCG...
   Leu Ala Ile Arg Cys Ser
Authenticity test: Restriction enzyme pattern analysed at GeneCorner: AgeI/SpeI, HindIII, NdeI and PvuII.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: -
Plasmid reference: PhD thesis Corinne Urwyler
Restricted distribution: - BCCM MTA
Distributed as: 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)
Cultivation temperature: 28°C
Biosafety level: L1 in E. coli; L2 in mammalian cells
Other culture collection numbers: -

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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