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pCSII-CMV-MCS _Y (YFP- type) c-GRAPHIC T2A H2B-Azurite

Lentiviral expression vector of GRAPHIC probe (LZB).

Catalog number RDB15874
Resource name pCSII-CMV-MCS _Y (YFP- type) c-GRAPHIC T2A H2B-Azurite
Clone info. Lentiviral expression vector of GRAPHIC probe (GPI anchored Reconstitution-Activated Proteins Highlight Intercellular Connections), fluorescent indicator for intercellular contacts with optimized intercellular GFP reconstitution. This probe contains an mouse preproacrosin signal peptide (SP), superfolder GFP-CT (YFP type), basic leucine zipper domain (LZB), mouse Thy-1 GPI-anchored domain, 2A peptide, human histone 2B (H2B), Azurite, see Fig. 1 of Kinoshita, N. et al., iScience 15: 28-38, 2019.
Comment Expression was confirmed by the depositor with detection of reconstituted YFP fluorescence and nucleic Azurite fluorecence.
Vector backbone CSII-CMV-MCS (plasmid)
Size of vector backbone 8.6 kb
Selectable markers Ampicillin (E. coli). Please note that Zeo resistance marker is not included in the lentivirus produced from this vector.
Gene/insert name mouse Acr cDNA mouse Thy1 cDNA human histon H2BJ cDNA Aequorea victoria GFP cDNA
Depositor|Developer Shimogori, Tomomi | Kinoshita, Nagatoki |
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External Database
mouse Acr mouse Thy1 human histon H2BJ Aequorea victoria GFP

      Reference sequence
        
       
      Sequence (full) RDB15874zzt01.seq compiled by us
      Sequence (partial) RDB15874pka01.seq provided by the depositor

      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 for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR (Kinoshita, N. et al., iScience 15: 28-38 (2019)) and an acknowledgment to the DEPOSITOR are requested. The use of the BIOLOGICAL RESOURCE is restricted to the academic researches conducted by non-profit organization.
      Additional terms and conditions:
      The RECIPIENT agrees to expressly describe the late Dr. Hiroyuki Miyoshi as the Developer of the lentivirus vector.
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      Remarks Remember that you will be working with samples containing infectious virus.
      提供案内 (日本国内) [open/close]

      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 (Kinoshita, N. et al., iScience 15: 28-38 (2019)) を引用し、謝辞の表明を必要とする。本件リソースの使用は学術機関での学術研究に限る。
      付加的提供条件:
      レンチウイルスベクターについて、本件研究材料を利用した研究結果等を発表する際は、レンチウイルスベクターが故三好浩之博士により開発されたことを明示する。
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      提供同意書 (MTA, 非営利機関による非営利学術研究用)[Word]
      手続きの概要は、「提供申込みについて[link]」をご覧ください。
      備考 このリソースはウイルス粒子産生用のため、取扱いに注意が必要です。

      Catalog # Resource name Shipping form Fee (non-profit org.)
      RDB15874 pCSII-CMV-MCS _Y (YFP- type) c-GRAPHIC T2A H2B-Azurite DNA solution

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

      How to cite this biological resource

      Materials & Methods section:

      The pCSII-CMV-MCS _Y (YFP- type) c-GRAPHIC T2A H2B-Azurite was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15874).

      Reference section:

      Kinoshita, N., Huang, A.J.Y., McHugh, T.J., Suzuki, S.C., Masai, I., Kim, I.H., Soderling, S.H., Miyawaki, A., Shimogori, T., Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections. iScience 15: 28-38 (2019). PMID 31026667. [link to RRC of NBRP]

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


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      QC test results

      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 RDB15874_A8Cdp1-1.pdf check

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

      Primer: D05101#1 (Pr0993)
      Region: insert 5', CMV pro_2
      Sequence file: RDB15874_A8Cda.seq check
      >D05101A6_A8Cd_1_D05101#1_G02_20_ABI24.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNG GCGTTCTTCT TCTTNNGGGC TTGCAGCTTC
         61 TTCTTGAGCT GGGCCATGGT GGCGCTAGCC GATCCGCCAC CGCCAGAGCC ACCTCCGCCT
        121 GAACCGCCTC CACCTCTAGC CTTGTACAGC TCGTCCATGC CGAGAGTGAT CCCGGCGGCG
        181 GTCACGAACT CCAGCAGGAC CATGTGATCG CGCTTCTCGT TGGGGTCTTT GCTCAGGACG
        241 GTCTGGTAGC TCAGGTAGTG GTTGTCGGGC AGCAGCACGG GGCCGTCGCC GATGGGGGTG
        301 TTCTGCTGGT AGTGGTCGGC GAGCTGCACG CTGCCGTCCT CGACGTTGTG GCGGACCGTG
        361 AAGTTCGCCT TGATGCCGTT CTTTCTAGCC TGCAGCGTGG TGTTATCCTT GGCAACCACG
        421 GACACTGCCA AGACCAGCAC AGCGACAGTT GGCAGCATCT CTACCATGGT GGCTCTAGCG
        481 GATCTGACGG TTCACTAAAC CAGCTCTGCT TATATAGACC TNNNNNNGTA CACGCCTACC
        541 GCCCATTTGC GTCAATGGGG CGGAGTTGTT ACGACATTTT GGAAAGTCCC GTTGATTTTG
        601 GTGCCAAAAC AAACTCCCAT TGACGTCAAT GGGGTGGAGA CTTGGAAAAT CCCCGTGAGT
        661 CNAANNCGCT ATCCACGCCC ATTGATGTAC TGCCNANNNN NNCATCANNN TGGTAATAGC
        721 GATGACTAAT ACGTNNNATG NNACTGNCNN AGNANGAAAG TCCCNNNAAN NGGTCATGTA
        781 CTNNNNATAA TGCCANGCGG NNCATTTNNC CGTCATNNNN NGTNCANTNN NGGGCGNACT
      841 TNNCATATGN NNACACTTGA NGNNACTGNN NNNNNNNN
      //
      Primer: PRE_rev (Pr0250)
      Region: PRE, insert 3'
      Sequence file: RDB15874_A8Cdb.seq check
      >D05101A6_1_A8Cd_PRE_rev_C01_03_ABI08.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNANNNNNNN NNNATAGTTA AGANNACNNG TCAATCTTTC
         61 NCAAATTTTG TAATCCAGAG GTTGATTATC GATAAGCTTG ATCCCTCGAT GTTAACTCTA
        121 GATTACTTGT ACAGCTCGTC CATGCCGTGA GTGATCCCGG CGGCGGTGCG GAACTCCAGC
        181 AGGACCATGT GATCGCGCTT CTCGTTGGGG TCTTTGCTCA GGGCGGACTG GGTGCTCAGG
        241 TAGTGGTTGT CGGGCAGCAG CACGGGGCCG TCGCCGATGG GGGTGTTCTG CTGGTAGTGG
        301 TCGGCGAGCT GCACGCTGCC GTCCTCGATG TTGTGGCGGA TCTTGAAGTT CACCTTGATG
        361 CCGTTCTTCT GCTTGTCGGC CATGATATAG ATGTTGTGGC TGTTGAAGTT GTACTCCAGC
        421 TTGTGCCCCA GGATGTTGCC GTCCTCCTTG AAGTCGATGC CCTTCAGCTC GATGCGGTTC
        481 ACCAGGGTGT CGCCCTCGAA CTTCACCTCG GCGCGGGTCT TGTAGTTGCC GTCGTCCTTG
        541 AAGAAGATGG TGCGCTCCTG GACGTANCCT TCGGGCATGG CGGACTTGAA GAAGTCGTGC
        601 TGCTTCATGT GGTCGGGGTA GCGGCTGAAG CACTGCACGC CGTGGCTCAN GGTGGTCACG
        661 ANGGTGGGCC AGGGCACGGG CAGCTTGCCG GTGGTGCAGA TGAACTTCAG GGTCAGCTTG
        721 NCGTANGTGG CATCGCCCTC GCCCTCGCCG GACACGCTGA ACTTGTGGNC GTTTACGTCN
        781 CCNTCCNGCT CGACCANGAT GGGNACCNCC CNNGTGAANC AGCTCCTCGC CCTTGCTCAT
        841 GNATCCCTTA NCGCTGGTGN ACTNNGNGAT GNNNNNANTN NNNTCGGNNA CNGNCNNGCT
        901 NNGNNNACTN CCNNNNNNNN CNNCNNNNNC ANNNNNNNCN GGATNNNCCN NNNNNNNNAN
      961 NNNNNNNNN
      //
      Primer: D05101#2 (Pr0994)
      Region: insert mid
      Sequence file: RDB15874_A8Cdc.seq check
      >D05101A6_A8Cd_1_D05101#2_H02_23_ABI24.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN GNNGNGGNGG NNCNGCNAGC GCCNCCATGG
         61 CCNAGCTCNA GAAGAAGCTG CAAGCCNTGA AGAAGAAGAA CGCCCAGCTC AAGTGGAAGC
        121 TCCAGGCCCT GAAGAAGAAG CTGGCCCAGA AGGCTAGCGG TGGAGGCGGT TCAGGCGGAG
        181 GTGGCTCTGG CGGTGGCGGA TCGTCTAGCG GTGGAGGCGG TTCAGGCGGA GGTGGCTCTG
        241 GCGGTGGCGG ATCGTCTAGC GGTGGAGGCG GTTCAGGCGG AGGTGGCTCT GGCGGTGGCG
        301 GATCGTCTAG CTACCAACTC GAGAATGGCG GCATAAGCCT GCTGGTTCAG AACACATCCT
        361 GGATGCTGCT GCTGCTGCTT TCCCTCTCCC TCCTCCAAGC CCTGGACTTC ATTTCTCTGT
        421 CTAGCGAGGG CAGAGGAAGT CTTCTAACAT GCGGTGACGT CGAGGAGAAT CCTGGCCCAA
        481 TGCCAGAGCC AGCGAAGTCT GCTCCCGCCC CGAAAAAGGG CTCCAAGAAG GCGGTGACTA
        541 AGGCGCAGAA GAAAGGCGGC AAGAAGCGCA AGCGCAGCCG CAAGGAGAGC TATTCCATCT
        601 ATGTGTACAA GGTTCTGAAG CAGGTCCACC CTGACACCGG CATTTCGTCC AAGGCCATGG
        661 GCATCATGAA TTCGTTTGTG AACGACATTT TCGAGCGCAT CGCAGGTGAN GCTTCCCGCC
        721 TGGCGCATTA CAACAAGCGC TCGACCATCA CCTCCAGGGA GATCCAGACG GCCGTGCGCC
        781 TGCTGCTGCC TGGGGAGTTG GCCANNCACG CCGNGTCCGA GGGTACTAAN NNCATCACCA
        841 AGTACACCAN CGCTAANGGA TCCATGANCA AGGGCGANGA NCTGTTCNCN NGGGGNGGNG
        901 CCCATCCNGG NNCGAGCTGG ANNGCGNNGT AAANCNGNNN CNANGTNNAN CNNGTNCNNN
        961 NAGGGNNAAN GGNNNNNGNN NACCTANNNN NNGCTGANNN NNNAANNNNN NNNNGNNNNN
      1021 NNNNN
      //
      Primer: EGFP-N_R2 (Pr0905)
      Region: insert mid
      Sequence file: RDB15874_A8Cdd.seq check
      >D05101A6_A8Cd_1_EGFP-N_R2_A03_03_ABI24.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNCNCNCNNN NGCTCANGGN
         61 GGTCACGNNG GNGGGCCAGG GCNNCGGGCA GCTTGCCGGT GGTGCAGATG AACTTCAGGG
        121 TCAGCTTGCC GTAGGTGGCA TCGCCCTCGC CCTCGCCGGA CACGCTGAAC TTGTGGCCGT
        181 TTACGTCGCC GTCCAGCTCG ACCAGGATGG GCACCACCCC GGTGAACAGC TCCTCGCCCT
        241 TGCTCATGGA TCCCTTAGCG CTGGTGTACT TGGTGATGGC CTTAGTACCC TCGGACACGG
        301 CGTGCTTGGC CAACTCCCCA GGCAGCAGCA GGCGCACGGC CGTCTGGATC TCCCTGGAGG
        361 TGATGGTCGA GCGCTTGTTG TAATGCGCCA GGCGGGAAGC CTCACCTGCG ATGCGCTCGA
        421 AAATGTCGTT CACAAACGAA TTCATGATGC CCATGGCCTT GGACGAAATG CCGGTGTCAG
        481 GGTGGACCTG CTTCAGAACC TTGTACACAT AGATGGAATA GCTCTCCTTG CGGCTGCGCT
        541 TGCGCTTCTT GCCGCCTTTC TTCTGCGCCT TAGTCACCGC CNNNNTGGAG CCCTTTTTCG
        601 GGGCGGGAGC ANACTTCNCT GGCTCTGGCA TTGGGCCAGG ATTCTCCTCG ACGTCACCGC
        661 ATGTTAGAAN ACTTCCTCTG CCCTCGCTAN ACAGANAAAN NGANGTCNNN GGCTTGNANG
        721 AGGGANNAGG GAAANGNNNN CANCANCANC NTCCAGGANG TGNTCTGAAN NNNNNCANGC
        781 TNATNNCCGC CNTTCTCNAG TTGGTAGCTA GACGATCCGN NNCCGNCAGA GCCACCTCCN
        841 NNTGNNNCGC CTCNNNNNGC NAGACNATCC GCCACCGCCA NANCCNNCNT CCGNCCTGAN
        901 NCGCCTNCAC NGCTAGNNGA TCCGNCNCCG CCAGANNCAC CNNCNGCCNG NANCNGNNNN
        961 NACCGCNNNN NNNNTNNNGG GGNCANCTNN NTNNTTNNNN GGNCNNNGNN NNCNTNNNNN
      1021 NNTTNNNNNN NNGGNNNNTT NNNTTNNNNN NN
      //
      Primer: HIV-B_R (Pr0673)
      Region: HIV-1 psi, 5'LTR, CMV pro_1, CMV en_1
      Sequence file: RDB15874_A8Cde.seq check
      >D05101A6_A8Cd_1_HIV-B_R_B03_06_ABI24.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNTT TGGNGNANTC NNNNNNCGCC GCCCCTCGCC
         61 TCTTGCCGTG CGCGCTTCNN CAAGCCGAGT CCTGCGTCGA GAGAGCTCCT CTGGTTTCCC
        121 TTTCGCTTTC AAGTCCCTGT TCGGGCGCCA CTGCTAGAGA TTTTCCACAC TGACTAAAAG
        181 GGTCTGAGGG ATCTCTAGTT ACCAGAGTCA CACAACAGAC GGGCACACAC TACTTGAAGC
        241 ACTCAAGGCA AGCTTTATTG AGGCTTAAGC AGTGGGTTCC CTAGTTAGCC AGAGAGCTCC
        301 CAGGCTCAGA TCTGGTCTAA CCAGAGAGAC CCAGTACAGG CAAAACGCGC TGCTTATATA
        361 GACCTCCCAC CGTACACGCC TACCGCCCAT TTGCGTCAAT GGGGCGGAGT TGTTACGACA
        421 TTTTGGAAAG TCCCGTTGAT TTTGGTGCCA AAACAAACTC CCATTGACGT CAATGGGGTG
        481 GAGACTTGGA AATCCCCGTG AGTCAAACCG CTATCCACGC CCATTGATGT ACTGCCAAAA
        541 CCGCATCACC ATGGTAATAG CGATGACTAA TACGTAGATG TACTGCCAAG TAGGAAAGTC
        601 CCATAAGGTC ATGTACTGGG CATAATGCCA GGCGGGCCAT TTANCGTCAT TGACGTCAAT
        661 AGGGGGCGTA CTTGGCATAT GATACACTTG ATGTACTGCC AAGTGGGCAG TTTACCGTAA
        721 ATACTCCACC CATTGACGTC AATGGAAAGT CNCTATTGGC GTTACTATGG GGAACATACG
        781 TCATTATTGA CGTCNATGGG CGGGGGTCGT NGGGCNGGTC AGCCAGGCGG NNCATTTANC
        841 GTANNTTATG TAACGCGGAA CTNNNTATAT GGGCTATGAN CTAATGACCC CGNAATTGAT
        901 TACTATNNAN NACNANNCNA NNANTCNANG NNNAACNNNN NNATCNGGNN NNNNNNTNNN
      961 CGAANNNNNN NNAANNNNNN NNNN
      //
      Primer: BGH_rev2 (Pr0606)
      Region: BGH pA, 3'LTR
      Sequence file: RDB15874_A8Cdf.seq check
      >D05101A6_A8Cd_1_BGH_rev2_E01_05_ABI08.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNGGGNN NNNNNNNNNN NNNNGANGGC TGNNNNNNNG
         61 AAGGCACNNN NNAGGCTGAT CAGCGGGTTT AAACGGGCCC TGCTAGAGAT TTTCCACACT
        121 GACTAAAAGG GTCTGAGGGA TCTCTAGTTA CCAGAGTCAC ACAACAGACG GGCACACACT
        181 ACTTGAAGCA CTCAAGGCAA GCTTTATTGA GGCTTAAGCA GTGGGTTCCC TAGTTAGCCA
        241 GAGAGCTCCC AGGCTCAGAT CTGGTCTAAC CAGAGAGACC CAGTACAGTC CGGATGCAGC
        301 TCTCGGGCCA TGTGATGAAA TGCTAGGCGG CTGTCAAACC TCCACTCTAA TACTTCTCTC
        361 TCCGGGTCAT CCATCCCATG CAGGCTCACA GGGTGTAACA AGCGGGTGTT CTCTCCTTCA
        421 TTGGCTTCTT CTACCTTCTC TTGCTCAACT GGTACTAGCT TGTAGCACCA TCCAAAGGTC
        481 NNNGGATATC TGATCCCTGG CCCTGGTGTG TAGTTCTGCC AATCAGGGAA GTANCCTTGT
        541 GTGTGGTAGA TCCNCAGATC AAGGATATCT TGTCTTCNTT GGGAGTGAAT TAGCCCTTCC
        601 AGTCCCCCCT TTTCTTTTAA AAAGTGGCTN ANANCTACAG CTGCCTTGNN AGTCNNTGGN
        661 CTTAAAGGNA NNTGANGNGT GACTGNNAAA CCCACCTCCT NCTNCNCTTG NGNTTCTANC
        721 CNNGCNCNAN TCNNNNTNGG NANCTGCNGN ANNNGNNNCN TGTNATNGNT CNNNGTTTTN
      781 NNNNNNNNN
      //
      Primer: SV40pro_F (Pr0110)
      Region: EM7 pro, BleoR, SV40 pA
      Sequence file: RDB15874_A8Cdg.seq check
      >D05101A6_1_A8Cd_SV40pro_F_E01_05_ABI08.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNGCTT TTTTGGNNNG CCNNNGNTTT
         61 TGCAANANNN TCCCGGGAGC TTNNNNNNNN NNNNNNNNAT CTGATCAGCA CGTGTTGACA
        121 ATTAATCATC GGCATAGTAT ATCGGCATAG TATAATACGA CAAGGTGAGG AACTAAACCA
        181 TGGCCAAGTT GACCAGTGCC GTTCCGGTGC TCACCGCGCG CGACGTCGCC GGAGCGGTCG
        241 AGTTCTGGAC CGACCGGCTC GGGTTCTCCC GGGACTTCGT GGAGGACGAC TTCGCCGGTG
        301 TGGTCCGGGA CGACGTGACC CTGTTCATCA GCGCGGTCCA GGACCAGGTG GTGCCGGACA
        361 ACACCCTGGC CTGGGTGTGG GTGCGCGGCC TGGACGAGCT GTACGCCGAG TGGTCGGAGG
        421 TCGTGTCCAC GAACTTCCGG GACGCCTCCG GGCCGGCCAT GACCGAGATC GGCGAGCAGC
        481 CGTGGGGGCG GGAGTTCGCC CTGCGCGACC CGGCCGGCAA CTGCGTGCAC TTCGTGGCCG
        541 AGGAGCAGGA CTGACACGTG CTACGAGATT TCGATTCCAC CGCCGCCTTC TATGAAAGGT
        601 TGGGCTTCGG AATCGTTTTC CGGGACGCCG GCTGGATGAT CCTCCNGCGC GGGGATCTCA
        661 TGCTGGAGTT CTTCGCCCAC CCCAACTTGT TTATTGCAGC TTATAATGGT TACNAATAAA
        721 GCAATAGCAT CACAAATTTC ACNAAATAAA GCATTTTTTT CACTGCATTC TAGTTGTGGT
        781 TTGTCCAAAC TCATCNATGT ATCTTATCAT GTCTGTATAC CGTCGANCTC TAGCTAGAGC
        841 TNGGCGTAAT CATGGNCATA GCTGTTNCCT GNNTGAAANT GNTANNCGNN NACNATTNCN
        901 NNNNNNATNN NNANNNGGNA NNNNNNNAAG NGNNAANNNN NNNNGNNNNC NNANNGANNN
      961 NNNCNNANNN NNNNNNN
      //

      Please visit Sequencing and PCR primers for primer information.


      References

      Original, user report and related articles

      original Kinoshita, N., Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections. iScience 15: 28-38 (2019). PMID 31026667. [link to RRC of NBRP]

      2024.03.26

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