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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/PlyA2-EGFP-His6

Marker of sphingomyelin and cholesterol lipid domains. Bacterial expression plasmid of EGFP-His6-tagged PlyA2.

Catalog number RDB13962
Resource name pET28/PlyA2-EGFP-His6
Clone info. Marker of sphingomyelin and cholesterol lipid domains. Bacterial expression plasmid of EGFP-His6-tagged PlyA2.
Comment Expression was confirmed by the depositor.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Km^r
Gene/insert name Pleurotus eryngii PlyA2 cDNA Aequorea victoria GFP cDNA
Depositor|Developer Kobayashi, Toshihide | Abe, Mitsuhiro |
Other clones in our bank

External Database
Pleurotus eryngii PlyA2
    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(s) designated by the DEPOSITOR is requested. (Bhat, HB., J Lipid Res. 54 (10): 2933-2943, 2013.)
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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 (Bhat, HB., J Lipid Res. 54 (10): 2933-2943, 2013.)。
            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])。

            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB13962 pET28/PlyA2-EGFP-His6 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

            Sequence (full) RDB13962zzt01.seq compiled by us

            Original reference

            original Bhat, H.B., Evaluation of aegerolysins as novel tools to detect and visualize ceramide phosphoethanolamine, a major sphingolipid in invertebrates. FASEB J. 29 (9): 3920-3934 (2015). PMID 26060215.
            original Bhat, H.B., Binding of a pleurotolysin ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains. J. Lipid Res. 54 (10): 2933-2943 (2013). PMID 23918047.

            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 RDB13962_A6Btp1-3.pdf check

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

            Primer: pBR322_BamHI_R
            Region: RBS, insert 5' (PlyA2, EGFP 5')
            Sequence file: RDB13962_A6Btd.seq check
            >D03144A5_A6Bt_1_pBR 322 BamHI R_D04_10.ab1
                1 NNNNNNNNNN NNNGNNTNNN NNTANGGNNN NTGTGAGCGG NNAACAATTC CCCTCTAGAA
               61 ATAATTTTGT TNAACTTTAA GAAGGAGATA TACCGGTACC ATGGCATACG CACAATGGGT
              121 CATCATCATC ATCCACAACG TCGGGTCCAA AGACGTGAAG ATCGTAAACC TCAAGCCTTC
              181 TTGGGGGAAG CTTCATGCCG ACGGCGACAA AGACACAGAA GTCTCAGCAA GCAAGTACGA
              241 AGGAACAGTC ATCAAGCCTG ACGAGAAGCT CCAGATCAAT GCCTGCGGCA GATCAGACGC
              301 CGCCGAAGGC ACGACAGGAA CCTTCGACCT TGTTGACCCC GCCGATGGCG ACAAGCAGGT
              361 CCGCCACTTC TACTGGGACT GCCCATGGGG AAGCAAGGCC AACACTTGGA CCGTTAGCGG
              421 TTCCAACACC AAGTGGATGA TCGAGTACTC TGGCCAGAAC TTGGACAGCG GTGCCCTCGG
              481 CACGATCACC GTCGACACCT TGAAGAAGGG GAACGAATTC GGAGGAGGAG GAGGAGGATC
              541 CATGGTGAGC AAGGGCGAGG AGCTGTTCAC CGGGGTGGTG CCCATCCTGG TCGAGCTGGA
              601 CGGCGACGTA AACGGCCACA AGTTCAGCGT GTCCGGCGAG GGCGAGGGCG ATGCCACCTA
              661 TGGCAAGCTG ANCCTGAAGT TCATCTGCAC CACCGGCAAG CTGCCCGTGC CCTGGCCCAC
              721 CCTCGTGACC ACCCTGACCT ACGGCGTGCA GTGCTTCAGC CGCTACCCCG ACCACATGAA
              781 GCAGCACGAC TTCTTCNAGT CCGCCATGCC CGAANGCTAC GTCCAGGANC GCANCATCTT
              841 CTTCNNNGAC GANNGNNACT ACAAGANCCG CGCCNANGTG NANTTCGNNG GCGANNCCCN
              901 GNNNNNNCNC ATCNAGCTGN ANNGNNNTCN NNTNNNNNNN NNANNNNNNC ATNCNGGGGN
            961 ANNNGCTGGN NTNNNNCTNN NNNNNNNN
            //
            Primer: EGFP-N_F
            Region: insert 3' (EGFP 3'), 6xHis
            Sequence file: RDB13962_A6Bte.seq check
            >13962_13962_B2D1F_1_EGFP-N_F_A01_01_ABI08.ab1
                1 NNNNNNNNNN NGNNNNNNNG NNGGCGNNGC NNCCTATGGC AAGCTGNNCC TGAAGTTCAT
               61 CTGCACCACC GGCAAGCTGC CCGTGCCCTG GCCCACCCTC GTGACCACCC TGACCTACGG
              121 CGTGCAGTGC TTCAGCCGCT ACCCCGACCA CATGAAGCAG CACGACTTCT TCAAGTCCGC
              181 CATGCCCGAA GGCTACGTCC AGGAGCGCAC CATCTTCTTC AAGGACGACG GCAACTACAA
              241 GACCCGCGCC GAGGTGAAGT TCGAGGGCGA CACCCTGGTG AACCGCATCG AGCTGAAGGG
              301 CATCGACTTC AAGGAGGACG GCAACATCCT GGGGCACAAG CTGGAGTACA ACTACAACAG
              361 CCACAACGTC TATATCATGG CCGACAAGCA GAAGAACGGC ATCAAGGTGA ACTTCAAGAT
              421 CCGCCACAAC ATCGAGGACG GCAGCGTGCA GCTCGCCGAC CACTACCAGC AGAACACCCC
              481 CATCGGCGAC GGCCCCGTGC TGCTGCCCGA CAACCACTAC CTGAGCACCC AGTCCGCCCT
              541 GAGCAAAGAC CCCAACGAGA AGCGCGATCA CATGGTCCTG CTGGAGTTCG TGACCGCCGC
              601 CGGGATCACT CTCGGCATGG ACGAGCTGTA CAAGAAGCTT CACCACCACC ACCACCACTG
              661 AGATCCGGCT GCTAACAAAG CCCGAAAGGA AGCTGANTTG GCTGCTGCCA CCGCTGAGCA
              721 ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT TGCTGANAGG
              781 AGGAACTATA TCCGGANTGN CGAATGGGAC GCGCCCNGTA GCGGCGCATT AAGCGCGGCG
              841 GGTGTGGTGG TTACGCGCAG CGNGACCGCT ACACTTGCCA GCGCCCTANC GCCCGCTNCT
              901 TTCGCTTTCN NCCCTTNCTT NCNCGCNNCN TNCGCCNGCT TNCCCCGNNN NGCTCNAANN
            961 CNGGGGNNNN CNNTNNNGNN NNNNNNTNNN NNNNTNNNNN NNNNN
            //
            Primer: EGFP-M_R
            Region: insert mid (EGFP 5', PlyA2 3')
            Sequence file: RDB13962_A6Btf.seq check
            >13962_13962_B2D1F_1_EGFP-M_R_A12_01_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNGGCGGNNC TNNNNNNNNC CTTGATGCCG TTCTTCTGCT
               61 TGTCGGCCAT GATATAGACG TTGTGGCTGT TGTAGTTGTA CTCCAGCTTG TGCCCCAGGA
              121 TGTTGCCGTC CTCCTTGAAG TCGATGCCCT TCAGCTCGAT GCGGTTCACC AGGGTGTCGC
              181 CCTCGAACTT CACCTCGGCG CGGGTCTTGT AGTTGCCGTC GTCCTTGAAG AAGATGGTGC
              241 GCTCCTGGAC GTAGCCTTCG GGCATGGCGG ACTTGAAGAA GTCGTGCTGC TTCATGTGGT
              301 CGGGGTAGCG GCTGAAGCAC TGCACGCCGT AGGTCAGGGT GGTCACGAGG GTGGGCCAGG
              361 GCACGGGCAG CTTGCCGGTG GTGCAGATGA ACTTCAGGGT CAGCTTGCCA TAGGTGGCAT
              421 CGCCCTCGCC CTCGCCGGAC ACGCTGAACT TGTGGCCGTT TACGTCGCCG TCCAGCTCGA
              481 CCAGGATGGG CACCACCCCG GTGAACAGCT CCTCGCCCTT GCTCACCATG GATCCTCCTC
              541 CTCCTCCTCC GAATTCGTTC CCCTTCTTCA AGGTGTCGAC GGTGATCGTG CCGAGGGCAC
              601 CGCTGTCCAA GTTCTGGCCA GANTACTCGA TCATCNACTT GGTGTTGGAA CCGCTNACGG
              661 TCCAAGTGNT GGNCTTGCTN CCCCNTGGGN AGTCCNNGTA NANNNGGNNN ANNNGCTNNN
            721 CGNNNTCGGC GGGNNNNNNN NNN
            //
            Primer: pGL3_RV4
            Region: lacI 3'
            Sequence file: RDB13962_A6Btg.seq check
            >D03144A5_A6Bt_1_pGL3_RV4_G12_21.ab1
                1 NNNNNNNNNN NNNNNNGNTC TCAGGGCNTC GGTCGANNNC CCGGTGCCTA ATGAGTGAGC
               61 TAACTTACAT TAATTGCGTT GCGCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC
              121 CAGCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG GTTTGCGTAT TGGGCGCCAG
              181 GGTGGTTTTT CTTTTCACCA GTGAGACGGG CAACAGCTGA TTGCCCTTCA CCGCCTGGCC
              241 CTGAGAGAGT TGCAGCAAGC GGTCCACGCT GGTTTGCCCC AGCAGGCGAA AATCCTGTTT
              301 GATGGTGGTT AACGGCGGGA TATAACATGA GCTGTCTTCG GTATCGTCGT ATCCCACTAC
              361 CGAGATATCC GCACCAACGC GCAGCCCGGA CTCGGTAATG GCGCGCATTG CGCCCAGCGC
              421 CATCTGATCG TTGGCAACCA GCATCGCAGT GGGAACGATG CCCTCATTCA GCATTTGCAT
              481 GGTTTGTTGA AAACCGGACA TGGCACTCCA GTCGCCTTCC CGTTCCGCTA TCGGCTGAAT
              541 TTGATTGCGA GTGAGATATT TATGCCAGCC AGCCAGACGC AGACGCGCCG AGACAGAACT
              601 TAATGGGCCC GCTAACAGCG CGATTTGCTG GTGANCCAAT GCGACCAGAT GCTCCACGCC
              661 CAGTCGCGTA CCGTCTTCAT GGGAGAAAAT AATACTGTTG ATGGGTGTCT GGTCAGANAC
              721 ATCAAGAAAT AACGCCNGAA CATTAGTGCA NGCAGCTTCC ACAGCAATGG CATCCTGGTC
              781 ATCCAGCGGA TAGTTAATGA TCAGCCCANN GANGCGTTGC GCGAGAAGNN TGTGCANCGC
              841 CGCTTTNCAG GCNTCNACGC CGCTTCNTTC TANCNTCNAC NCCACCACNC NNNNACCCAG
            901 TTGNNNNNNG NNANATTTNA TCGNNNNNAN NNNTTGCNAN NNNNNNNNN
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Bhat, H.B., Evaluation of aegerolysins as novel tools to detect and visualize ceramide phosphoethanolamine, a major sphingolipid in invertebrates. FASEB J. 29 (9): 3920-3934 (2015). PMID 26060215.
            original Bhat, H.B., Binding of a pleurotolysin ortholog from Pleurotus eryngii to sphingomyelin and cholesterol-rich membrane domains. J. Lipid Res. 54 (10): 2933-2943 (2013). PMID 23918047.
            protocol Abe, M., Dynamics of sphingomyelin-and cholesterol-enriched lipid domains during cytokinesis. Methods Cell Biol. 137: 15-24 (2017). PMID 28065303.

            2022.05.17

            GNP_filter3_RDBDEP_html_220516.pl