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Please review the QC test results indicated by check icon below as well as clone information before placing your order.

pET28/His6-EGFP-NT-Lys

Marker of sphingomyelin in cell membrane. Bacterial expression plasmid of nontoxic Lysenin.

Catalog number RDB13498
Resource name pET28/His6-EGFP-NT-Lys
Clone info. Marker of sphingomyelin in cell membrane. Bacterial expression plasmid of nontoxic Lysenin.
Comment 169-297 AA mutant. Reference of DNA sequence, D85846.1.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Kan^r
Gene/insert name common brandling worm Lysenin cDNA Aequorea victoria GFP cDNA
Depositor|Developer Kobayashi, Toshihide | Abe, Mitsuhiro |
Other clones in our bank

External Database
common brandling worm Lysenin
    Aequorea victoria GFP

            Reference sequence
              

            Distribution information

            Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
            Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested. (Makino, A., FASEB J. 29 (2): 477-493, 2015)
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            Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
            This RESOURCE contains a fluorescent protein owned or licensed by Global Life Sciences Solutions Operations UK Ltd. (Cytiva). Please refer to Schedule B and use Schedule A (download the Schedule A and Schedule B [link]).
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            提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する(Makino, A., FASEB J. 29 (2): 477-493, 2015)。
            提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。営利目的利用についてはお問い合わせください。
            提供依頼書 [Word]
            提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]
            GFPリソースを含みます。Schedule Bをご一読の上Schedule A (英語版のみ) の提出をお願いします (Schedule AとSchedule Bを入手[link])。

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            RDB13498 pET28/His6-EGFP-NT-Lys DNA solution

            Please review the QC test results indicated by check icon below as well as clone information before placing your order.

            References and tips

            Electronic file

            Original reference

            original Makino, A., Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis. FASEB J. 29 (2): 477-493 (2015). PMID 25389132.

            Further references such as user reports and related articles (go to bottom)

            Featured content

            Featured content Visualization of Organelles (English text)
            Featured content Sphingolipid signaling pathway (English text)

            Sequence information

            RIKEN BRC has sequenced portions of this material for quality test.
            Please review the QC test results indicated by check icon as well as clone information before placing your order.

            Test sheet RDB13498_A6Cap1-2.pdf check

            Nucleotide sequence of a portion of this resource (if available).

            Primer: T7_20bp
            Region: 6xHis,thrombin site,insert 5'(EGFP 5')
            Sequence file: RDB13498_A6Cad.seq check
            >13498_13498_A6Ca_4_T7_20bp_E05_14.ab1
                1 NNNNNNNNNN NNNNNNNNNN NTTTGTTNNN CTTTAAGAAG GAGNNATACC ATGGGCAGCA
               61 GCCATCATCA TCATCATCAC AGCAGCGGCC TGGTGCCGCG CGGCAGCCAT ATGGCTAGAG
              121 TGAGCAAGGG CGAGGAGCTG TTCACCGGGG TGGTGCCCAT CCTGGTCGAG CTGGACGGCG
              181 ACGTAAACGG CCACAAGTTC AGCGTGTCCG GCGAGGGCGA GGGCGATGCC ACCTACGGCA
              241 AGCTGACCCT GAAGTTCATC TGCACCACCG GCAAGCTGCC CGTGCCCTGG CCCACCCTCG
              301 TGACCACCCT GACCTACGGC GTGCAGTGCT TCAGCCGCTA CCCCGACCAC ATGAAGCAGC
              361 ACGACTTCTT CAAGTCCGCC ATGCCCGAAG GCTACGTCCA GGAGCGCACC ATCTTCTTCA
              421 AGGACGACGG CAACTACAAG ACCCGCGCCG AGGTGAAGTT CGAGGGCGAC ACCCTGGTGA
              481 ACCGCATCGA GCTGAAGGGC ATCGACTTCA AGGAGGACGG CAACATCCTG GGGCACAAGC
              541 TGGAGTACAA CTACAACAGC CACAACGTCT ATATCATGGC CGACAAGCAG AAGAACGGCA
              601 TCAAGGTGAA CTTCAAGATC CGCCACAACA TCGAGGACGG CAGCGTGCAG CTCGCCGACC
              661 ACTACCAGCA GAACACCCCC ATCGGCGACG GCCCCGTGCT GCTGCCCGAC AACCACTACC
              721 TGANCACCCA GTCCGCCCTG ANCAAAGANC CCAACGAGAA GCGCGATCAC ATGGTCCTGC
              781 TGGANTTCGT GACCGCCGCC GGGATCACTC TCGGCATGNA CGANCTGTAC AAGAAGCTTA
              841 TAATCCTTGG AAAANCAGAA NNTCGAANTN AGCANGCAGA AANGAANGNN TACATGACAG
              901 TCGTTNCNAG AAAAAAGTTN GCCAGCNGCN NNNNTTGGNN ATANCAAACT TTTTCNNNNT
            961 NGNGNNNNAN NNNNNNNNN
            //
            Primer: T7 terminator
            Region: insert 3'(NT-Lys, EGFP 3')
            Sequence file: RDB13498_A6Cae.seq check
            >13498_13498_A6Ca_4_T7_terminator_F05_17.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNTNAGCAGC CGGNNCTCAG TGGTGGTGGT GGTGGTGCTC
               61 GAGTTAACCA ACCACTTCCA AAATCCACTT GTCTTCACGT TTGTCCAGAC AGTACACGTT
              121 TGTTGCCGGT CCATCATCGT AGCACAGCCC ACTGCGAGTG AAGTACTTAT TCATGAAGGT
              181 GACTACGTCA CCATGACGAA GAGGCAATGA CTTATTGATT GCCCAGCGCT GTTTCGGATT
              241 ATCAGTACCC TGATCAAAGT AGATTCCTCC TTGATCAGAG GAATAGGCAT ACTCATAGCC
              301 CGAATACATG GTGTTCAGCG TTTTAATTCG AAATCCCCCC CAATCTTCAT AGAGCACAAA
              361 CTTGAAAAGT TTGCTATGTC CAAGAGTTGC AGCTGGCCAA CTTTTTCTTG AAACGACTGT
              421 CATGTACTCC TTCCTTTCTG CATGCTTAAT TCGAATTTCT GTTTTTCCAA GGATTATAAG
              481 CTTCTTGTAC AGCTCGTCCA TGCCGAGAGT GATCCCGGCG GCGGTCACGA ACTCCAGCAG
              541 GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGG GCGGACTGGG TGCTCAGGTA
              601 GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT GGTAGTGGTC
              661 GGCGAGCTGC ACGCTGCCGT CCTCGATGTT GTGGCGGATC TTGAAGTTCA CCTTGATGCC
              721 GTTCTTCTGC TTGTCGGCCA TGATATAGAC GTTGTGGCTG TTGTAGTTGT ACTCCAGCTT
              781 GTGCCCCANG ATGTTGCCGT CCTCCTTGAA GTCNATGCNN TTCAGCTCGA TGCGGTTCAC
              841 CAGGGTGTCG CCCTCGAACT TCACNTCGGC GCNGGTCNTG TAGTTGNCGT CGTNCNNNAA
              901 GAANANGGTG CGCNNCNNGN NGTANCCNNC NGGNNTGGNG GNNTNGNANN AANNCGTGCT
            961 GCTNNNTGTN NNCGGGNNAN CGGNNGNNNN NNNNNN
            //
            Primer: T7 terminator
            Region: insert 3'(NT-Lys, EGFP 3')
            Sequence file: RDB13498_A6Cae.seq check
            >13498_13498_A6Ca_4_T7_terminator_F05_17.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNTNAGCAGC CGGNNCTCAG TGGTGGTGGT GGTGGTGCTC
               61 GAGTTAACCA ACCACTTCCA AAATCCACTT GTCTTCACGT TTGTCCAGAC AGTACACGTT
              121 TGTTGCCGGT CCATCATCGT AGCACAGCCC ACTGCGAGTG AAGTACTTAT TCATGAAGGT
              181 GACTACGTCA CCATGACGAA GAGGCAATGA CTTATTGATT GCCCAGCGCT GTTTCGGATT
              241 ATCAGTACCC TGATCAAAGT AGATTCCTCC TTGATCAGAG GAATAGGCAT ACTCATAGCC
              301 CGAATACATG GTGTTCAGCG TTTTAATTCG AAATCCCCCC CAATCTTCAT AGAGCACAAA
              361 CTTGAAAAGT TTGCTATGTC CAAGAGTTGC AGCTGGCCAA CTTTTTCTTG AAACGACTGT
              421 CATGTACTCC TTCCTTTCTG CATGCTTAAT TCGAATTTCT GTTTTTCCAA GGATTATAAG
              481 CTTCTTGTAC AGCTCGTCCA TGCCGAGAGT GATCCCGGCG GCGGTCACGA ACTCCAGCAG
              541 GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGG GCGGACTGGG TGCTCAGGTA
              601 GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT GGTAGTGGTC
              661 GGCGAGCTGC ACGCTGCCGT CCTCGATGTT GTGGCGGATC TTGAAGTTCA CCTTGATGCC
              721 GTTCTTCTGC TTGTCGGCCA TGATATAGAC GTTGTGGCTG TTGTAGTTGT ACTCCAGCTT
              781 GTGCCCCANG ATGTTGCCGT CCTCCTTGAA GTCNATGCNN TTCAGCTCGA TGCGGTTCAC
              841 CAGGGTGTCG CCCTCGAACT TCACNTCGGC GCNGGTCNTG TAGTTGNCGT CGTNCNNNAA
              901 GAANANGGTG CGCNNCNNGN NGTANCCNNC NGGNNTGGNG GNNTNGNANN AANNCGTGCT
            961 GCTNNNTGTN NNCGGGNNAN CGGNNGNNNN NNNNNN
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Makino, A., Visualization of the heterogeneous membrane distribution of sphingomyelin associated with cytokinesis, cell polarity, and sphingolipidosis. FASEB J. 29 (2): 477-493 (2015). PMID 25389132.
            protocol Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303.
            review Kobayashi, T., Impact of Intrinsic and Extrinsic Factors on Cellular Sphingomyelin Imaging with Specific Reporter Proteins.

            2022.05.17

            GNP_filter3_RDBDEP_html_220516.pl