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CSIV-RfA-TRE-EF-KT

Backbone vector plasmid of lentivirus construct driven by Tet-responsive promoter by Gateway(R) cloning with hKO1-2A-rtTA marker driven by EF-1 alpha promoter.

Catalog number RDB12872
Resource name CSIV-RfA-TRE-EF-KT
Clone info. Backbone vector plasmid of lentivirus construct driven by Tet-responsive promoter by Gateway(R) cloning with hKO1-2A-rtTA marker driven by EF-1 alpha promoter. Please note that the TRE and EF promoters are placed in back-to-back orientation and this construct contains loxP.
Comment Commonly requested with pCMV-VSV-G-RSV-Rev (RDB04393) and pCAG-HIVgp (RDB04394).
Vector backbone CSIV-RfA-TRE-EF-KT (Plasmid)
Selectable markers Ampicillin (E. coli). Please note that Zeo resistance marker is not included in the lentivirus produced from this vector.
Growth remarks Use DB3.1, ccdB Survival or equivalent to amplify this clone. 30 degC is fine.
Depositor|Developer Miyoshi, Hiroyuki |
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External Database

            Reference sequence
              
             
            Sequence (full) RDB12872zzt02.seq compiled by us

            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 The availability of the BIOLOGICAL RESOURCE is limited to a RECIPIENT of a not-for profit institution for a not-for-profit academic purpose. The RECIPIENT agrees to expressly describe the late Dr. Hiroyuki Miyoshi as the Developer.
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            Please visit Ordering instruction.[link] 
            Remarks Remember that you will be working with samples containing infectious virus.
            Use Survival2 (Invitrogen) or equivalent to amplify this clone.
            提供案内 (日本国内) [open/close]

            提供条件 本件研究材料は、非営利機関の非営利学術研究に限って提供する。本件研究材料を利用した研究結果等を発表する際は、本件研究材料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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            提供依頼書 [Word]
            提供同意書 (MTA, 非営利学術目的用)[Word]
            遺伝子組換え生物の受入れ確認書が必要です。当室にご請求ください。
            手続きの概要は、「レンチウイルスベクターの提供申し込み」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。
            このクローンの増殖にはSurvival2 (Invitrogen)あるいは同等の宿主を使用してください。

            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB12872 CSIV-RfA-TRE-EF-KT DNA solution

            Ordering instruction of plasmids [in Japanese] [in English]
            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 CSIV-RfA-TRE-EF-KT was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB12872).

            Reference section:

            Iida, T., Iwanami, A., Sanosaka, T., Kohyama, J., Miyoshi, H., Nagoshi, N., Kashiwagi, R., Toyama, Y., Matsumoto, M., Nakamura, M., Okano, H., Whole-Genome DNA Methylation Analyses Revealed Epigenetic Instability in Tumorigenic Human iPS Cell-Derived Neural Stem/Progenitor Cells. Stem Cells 35 (5): 1316-1327 (2017). PMID 28142229. [link to RRC of NBRP]

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


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            Featured content

            Featured content Lentiviral plasmid list (English text)
            Featured content Lentivirus Vector (English text)
            Featured content Lentivirus Vector (Japanese text)

            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 RDB12872_A4Krp1-3.pdf check

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

            Primer: EF1a-N_R (Pr0024)
            Region: EF1 alpha pro 5', tight TRE pro, attR1, lac UV5 pro
            Sequence file: RDB12872_A4Kri.seq check
            >12872_12872_B1HjF_2_EF1a-N_R_A03_01_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNGG CGNNGTGCGC TCTGCCNNCT GNCGGGCACC GGNNCCTCAC
               61 GAAGCTTGAA TTCGGCCCTT TCGTCTTCAA TCGAGTTTAC TCCCTATCAG TGATAGAGAA
              121 CGTATGTCGA GTTTACTCCC TATCAGTGAT AGAGAACGAT GTCGAGTTTA CTCCCTATCA
              181 GTGATAGAGA ACGTATGTCG AGTTTACTCC CTATCAGTGA TAGAGAACGT ATGTCGAGTT
              241 TACTCCCTAT CAGTGATAGA GAACGTATGT CGAGTTTATC CCTATCAGTG ATAGAGAACG
              301 TATGTCGAGT TTACTCCCTA TCAGTGATAG AGAACGTATG TCGAGGTAGG CGTGTACGGT
              361 GGGAGGCCTA TATAAGCAGA GCTCGTTTAG TGAACCGTCA GATCGCCGCT AGCATCACAA
              421 GTTTGTACAA AAAAGCTGAA CGAGAAACGT AAAATGATAT AAATATCAAT ATATTAAATT
              481 AGATTTTGCA TAAAAAACAG ACTACATAAT ACTGTAAAAC ACAACATATC CAGTCACTAT
              541 GGCGGCCGCA TTAGGCACCC CAGGCTTTAC ACTTTATGCT TCCGGCTCGT ATAATGTGTG
              601 GATTTTGAGT TAGGATCCGT CGAGATTTTC NGGAGCTAAG GAAGCTAAAA TGGAGAAAAA
              661 AATCACTGGA TATACCACCG TTGATATATC CCAATGGCAT CGTAAAGAAC ATTTTGAGGC
              721 ATTTCAGTCA GTTGCTCAAT GTACCTATAA CCAGACCGTT CAGCTGGATA TTACGGCNNT
              781 TTTTTAAAGA CCGNNAAAGA AAANTAANNA CAAGTTTTAT CNNGNCTTNN NNTCACNTTC
              841 TTGCCCGCCT GATGNANNGC TCATCCNNGN ANTTCCGNNN NNNNNNGAAN NNANGGGTGA
              901 ANCNGGNGNT NNNNGGNNNT NAGTGNTTCA NNNNNNTGNT TANNNNGNTT TTNCANGANC
            961 NAANNNNNNN NNGNTTTTCN TCNCNNNTGN AANNNGAATN NNNNNNNNN
            //
            Primer: EF1a-C_F (Pr0168)
            Region: EF1 alpha pro, hKO,
            Sequence file: RDB12872_A4Krj.seq check
            >12872_12872_A4Kr_2_EF1a-C_F_B02_05_ABI24.ab1
                1 NNNNNNNNNN NTTTTGNNNT NGGNNCTTGG TTNNTTCTCA AGCCTCAGAC AGTGGTTCAA
               61 AGTTTTTTTC TTCCATTTCA GGTGTCGTGA ACACGCTACC GGTCCACCAT GGTGAGCGTG
              121 ATCAAGCCCG AGATGAAGAT GAAGTACTTC ATGGACGGCA GCGTGAACGG CCACGAGTTC
              181 ACCGTGGAGG GCGAGGGCAC CGGCAAGCCC TACGAGGGCC ACCAGGAGAT GACCCTGAGG
              241 GTGACAATGG CCAAGGGCGG CCCCATGCCC TTCAGCTTCG ACCTGGTGAG CCACACCTTC
              301 TGCTACGGCC ACAGGCCCTT CACCAAGTAC CCCGAGGAGA TCCCCGACTA CTTCAAGCAG
              361 GCCTTCCCCG AGGGCCTGAG CTGGGAGAGG AGCCTCCAGT TCGAGGACGG CGGCTTCGCC
              421 GCCGTGAGCG CCCACATCAG CCTGAGGGGC AACTGCTTCG AGCACAAGAG CAAGTTCGTG
              481 GGCGTGAACT TCCCCGCCGA CGGCCCCGTG ATGCAGAACC AGAGCAGCGA CTGGGAGCCC
              541 AGCACCGAGA AGATCACCAC CTGCGACGGC GTGCTGAAGG GCGACGTGAC CATGTACCTG
              601 AAGCTGGCCG GCGGCGGCAA CCACAAGTGC CAGTTCAAGA NCACCTACAA GGCCGCCAAG
              661 AAGATCCTGA AGANGCCCCA GANCCACTTC ATCGGCCACA GGCTGGTGAG GAAGACCNAN
              721 GGCNACATCA CCGANCTGGT GGANGANGCC NNGGCCCACN GCGANGGCAN ANGAAGTCTT
              781 CTAACATGCN GTGACGTGGN NNANANTCCC GGCCCNATGT CTAGACNGNN NNAGANNNAA
              841 GTCNTNAANN GCGCTCNNGN NNNTNNNTCN NNNGNNNNCN GNATNNNANN NNCNGANNAN
            901 NNNNNAACNN NNNNNAANNN NNNGNNNNTN NNNNNNNNN
            //
            Primer: tTA-Ad R (Pr0542)
            Region: rtTA-Advanced 3',T2A,hKO
            Sequence file: RDB12872_A4Krk.seq check
            >12872_12872_B1HjF_tTA-AdR_E09_05_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNN NGANTTTGCT CTTGTCNNGT CTAGACATAG GGCCGGGATT
               61 CTCCTCCACG TCACCGCATG TTAGAAGACT TCCTCTGCCC TCGCAGTGGG CCACGGCGTC
              121 CTCCACCAGC TCGGTGATGT TGCCCTCGGT CTTCCTCACC AGCCTGTGGC CGATGAAGTG
              181 GCTCTGGGGC ATCTTCAGGA TCTTCTTGGC GGCCTTGTAG GTGGTCTTGA ACTGGCACTT
              241 GTGGTTGCCG CCGCCGGCCA GCTTCAGGTA CATGGTCACG TCGCCCTTCA GCACGCCGTC
              301 GCAGGTGGTG ATCTTCTCGG TGCTGGGCTC CCAGTCGCTG CTCTGGTTCT GCATCACGGG
              361 GCCGTCGGCG GGGAAGTTCA CGCCCACGAA CTTGCTCTTG TGCTCGAAGC AGTTGCCCCT
              421 CAGGCTGATG TGGGCGCTCA CGGCGGCGAA GCCGCCGTCC TCGAACTGGA GGCTCCTCTC
              481 CCAGCTCAGG CCCTCGGGGA AGGCCTGCTT GAAGTAGTCG GGGATCTCCT CGGGGTACTT
              541 GGTGAANGGC CTGTGGCCGT ANCANAANGT GTGNNNNACC ANGTCGAANC TGANNGGCNT
              601 GGGGCCGCNC TTGGNCNNNG TCACCCTNNN NGTCATCTCC NGNNGNCCCT CGTANGNNNN
            661 GNNNGTGCCC NNNNNNNNNN
            //
            Primer: CMV_Forward (Pr0016)
            Region: CMV pro, 3', 5' LTR, HIV-1 psi
            Sequence file: RDB12872_A4Krl.seq check
            >12872_12872_B1HjF_2_CMV_Forward_E03_05_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNTG NNGCNANNGG GCGGTANGCG TGTACGGTGG GAGGTCTATA
               61 TAAGCAGCGC GTTTTGCCTG TACTGGGTCT CTCTGGTTAG ACCAGATCTG AGCCTGGGAG
              121 CTCTCTGGCT AACTAGGGAA CCCACTGCTT AAGCCTCAAT AAAGCTTGCC TTGAGTGCTT
              181 CAAGTAGTGT GTGCCCGTCT GTTGTGTGAC TCTGGTAACT AGAGATCCCT CAGACCCTTT
              241 TAGTCAGTGT GGAAAATCTC TAGCAGTGGC GCCCGAACAG GGACTTGAAA GCGAAAGGGA
              301 AACCAGAGGA GCTCTCTCGA CGCAGGACTC GGCTTGCTGA AGCGCGCACG GCAAGAGGCG
              361 AGGGGCGGCG ACTGGTGAGT ACGCCAAAAA TTTTGACTAG CGGAGGCTAG AAGGAGAGAG
              421 ATGGGTGCGA GAGCGTCAGT ATTAAGCGGG GGAGAATTAG ATCGCGATGG GAAAAAATTC
              481 GGTTAAGGCC AGGGGGAAAG AAAAAATATA AATTAAAACA TATAGTATGG GCAAGCAGGG
              541 AGCTAGAACG ATTCGCAGTT AATCCTGGCC TGTTAGAAAC ATCAGAAGGC TGTAGACAAA
              601 TACTGGGACA GCTACAACCA TCCCTTCAGA CAGGATCANA NNAACTTAGA TCNNTATATA
              661 ATACAGTANC NACCCTCTAT TGTGTGCATC NAAAGGATAG AGATAAAAGA CACCAAGGAA
              721 NGCTTTANAC AANNNNNNNN GAAGAGCAAA ACNNNGTAAG ACCNCCGCAC AGCANNCGGN
              781 CNGCTGATCN TNNANANNTG GANNGNNNNN NTATGANGGN ACAATTGGNA GAANNNGNNN
              841 TTANNTNNAN NNNANANNNN NTANANNNNT NGAANCNANN TNNNNNNNNA NNNNNNNNNN
            //
            Primer: PRE_rev (Pr0250)
            Region: PRE,rtTA-Advanced 3'
            Sequence file: RDB12872_A4Krm.seq check
            >12872_12872_B1HjF_PRE_rev_G01_19_ABI24.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNTA GTTAAGAATA CCAGTCAATC TTTCACAAAT
               61 TTTGTAATCC AGAGGTTGAT TATCGATAAG CTTGATCCCT CGATGTTAAC TCTAGAGGAT
              121 CCTTACTTAG TTACCCGGGG AGCATGTCAA GGTCAAAATC GTCAAGAGCG TCAGCAGGCA
              181 GCATATCAAG GTCAAAGTCG TCAAGGGCAT CGGCTGGGAG CATGTCTAAG TCAAAATCGT
              241 CAAGGGCGTC GGCCGGCCCG CCGCTTTCGC ACTTTAGCTG TTTCTCCAGG CCACATATGA
              301 TTAGTTCCAG GCCGAAAAGG AAGGCAGGTT CGGCTCCCTG CCGGTCGAAC AGCTCAATTG
              361 CTTGTCTCAG AAGTGGGGGC ATAGAATCGG TGGTAGGTGT CTCTCTTTCC TCTTTTGCTA
              421 CTTGATGCTC CTGTTCCTCC AATACGCAGC CCAGTGTAAA GTGGCCCACG GCGGACAGAG
              481 CGTACAGTGC GTTCTCCAGG GAGAAGCCTT GCTGACACAG GAACGCGAGC TGATTTTCCA
              541 GGGTTTCGTA CTGTTTCTCT GTTGGGCGGG TGCCGAGATG CACTTTAGCC CCGTCGCGAT
              601 GTGAGAGGAG AGCACAGCGG ANNGACTTGN CGTTGTTCCG CAGAAAGTCT TGCCNTGACT
              661 CGCCTTCCAG GGGGCAGAAN TGGGTATGAT GCCNGTCCAG CATCTCGNNN NNNNNGGNNA
            721 TCNNNNNNGN NCNNNNTNNN NNNNNNN
            //
            Primer: bGH_rev3 (Pr0885)
            Region: bGH pA 5', 3' LTR, loxP
            Sequence file: RDB12872_A4Krn.seq check
            >12872_12872_B1HjF_2_bGH_rev3_F03_06_ABI08.ab1
                1 NNNNNNNNNN NNNNGNNNNN NNGGNNGGGC ANNNNNNGNN GGCTGGCAAC TAGAAGGCAC
               61 AGTCGAGGCT GATCAGCGGG TTTAAACGGG CCCTGCTAGA GATTTTCCAC ACTGACTAAA
              121 AGGGTCTGAG GGATCTCTAG TTACCAGAGT CACACAACAG ACGGGCACAC ACTACTTGAA
              181 GCACTCAAGG CAAGCTTTAT TGAGGCTTAA GCAGTGGGTT CCCTAGTTAG CCAGAGAGCT
              241 CCCAGGCTCA GATCTGGTCT AACCAGAGAG ACCCAGTACA GTCCGGAATA ACTTCGTATA
              301 ATGTATGCTA TACGAAGTTA TGATATCTTG TCTTCGTTGG GAGTGAATTA GCCCTTCCAG
              361 TCCNNNNNNT TCTTNNNNNN AGTGGCTAAG ATCTACAGCT GCCTTGTAAG TCATTGGTCT
              421 TAAAGGTACC TGAGGTGTGA CTGGAAAACC CACCTCCTCC TCCTCTTGTG CTTCTAGCCA
              481 NGCNCAANCN NNNTTGGTAG CNNNNNTATT GNNNNNTGNG NNTGCTCCAT GTTTTTCTAG
              541 GTNNNNNNGG TATCNATGCN GGGAGGCGGN NNNNANGGNN NNNNNANTCN NCTGAGGGCN
            601 NNNGNNNNNN CNCNNANNAN GCNNNNNANN NNNNNNNN
            //
            Primer: SV40pro_ori_F (Pr0731)
            Region: SV40 pro_ori, Zeo
            Sequence file: RDB12872_A4Kro.seq check
            >12872_12872_B1HjF_2_SV40pro_ori_F_G03_07_ABI08.ab1
                1 NNNNNNNNNN NNNNGNNGCC NCNGCCTCTG NNCTANTCNN GAAGTAGTGA GGAGGCTTTT
               61 TTGGAGGCCT AGGCTTTTGC AAAAAGCTCC CGGGAGCTTG TATATCCATT TTCGGATCTG
              121 ATCAGCACGT GTTGACAATT AATCATCGGC ATAGTATATC GGCATAGTAT AATACGACAA
              181 GGTGAGGAAC TAAACCATGG CCAAGTTGAC CAGTGCCGTT CCGGTGCTCA CCGCGCGCGA
              241 CGTCGCCGGA GCGGTCGAGT TCTGGACCGA CCGGCTCGGG TTCTCCCGGG ACTTCGTGGA
              301 GGACGACTTC GCCGGTGTGG TCCGGGACGA CGTGACCCTG TTCATCAGCG CGGTCCAGGA
              361 CCAGGTGGTG CCGGACAACA CCCTGGCCTG GGTGTGGGTG CGCGGCCTGG ACGAGCTGTA
              421 CGCCGAGTGG TCGGAGGTCG TGTCCACGAA CTTCCGGGAC GCCTCCGGGC CGGCCATGAC
              481 CGAGATCGGC GAGCAGCCGT GGGGGCGGGA GTTCGCCCTG CGCGACCCGG CCGGCAACTG
              541 CGTGCACTTC GTGGCCGAGG AGCAGGACTG ACACGTGCTA CGAGATTTCG ATTCCACCGC
              601 CGCCTTCTAT GAAAGGTTGG GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT
              661 CCAGCGCGGG GATCTCATGC TGGANTTCTT CGCCCACCNN NNNTTGTTTA NTGCANCNTT
              721 ATNANTGGTT ACNAATAAAA GCAANNAGCA TCANNAATTT TCACAAAATA AAGCATTTTT
              781 TTCACTGCAT NCNANNTTNG NGGNTTGTCC AAACTCATCA ANGNNNNNNN NNCANGTNNG
            841 NNNNACNNNN NNNNNN
            //
            Primer: SV40pro_ori_F (Pr0731)
            Region: SV40 pro_ori, Zeo
            Sequence file: RDB12872_A4Krp.seq check
            >12872_12872_B1HjF_2_SV40pro_ori_F_G03_07_ABI08.ab1
                1 NNNNNNNNNN NNNNGNNGCC NCNGCCTCTG NNCTANTCNN GAAGTAGTGA GGAGGCTTTT
               61 TTGGAGGCCT AGGCTTTTGC AAAAAGCTCC CGGGAGCTTG TATATCCATT TTCGGATCTG
              121 ATCAGCACGT GTTGACAATT AATCATCGGC ATAGTATATC GGCATAGTAT AATACGACAA
              181 GGTGAGGAAC TAAACCATGG CCAAGTTGAC CAGTGCCGTT CCGGTGCTCA CCGCGCGCGA
              241 CGTCGCCGGA GCGGTCGAGT TCTGGACCGA CCGGCTCGGG TTCTCCCGGG ACTTCGTGGA
              301 GGACGACTTC GCCGGTGTGG TCCGGGACGA CGTGACCCTG TTCATCAGCG CGGTCCAGGA
              361 CCAGGTGGTG CCGGACAACA CCCTGGCCTG GGTGTGGGTG CGCGGCCTGG ACGAGCTGTA
              421 CGCCGAGTGG TCGGAGGTCG TGTCCACGAA CTTCCGGGAC GCCTCCGGGC CGGCCATGAC
              481 CGAGATCGGC GAGCAGCCGT GGGGGCGGGA GTTCGCCCTG CGCGACCCGG CCGGCAACTG
              541 CGTGCACTTC GTGGCCGAGG AGCAGGACTG ACACGTGCTA CGAGATTTCG ATTCCACCGC
              601 CGCCTTCTAT GAAAGGTTGG GCTTCGGAAT CGTTTTCCGG GACGCCGGCT GGATGATCCT
              661 CCAGCGCGGG GATCTCATGC TGGANTTCTT CGCCCACCNN NNNTTGTTTA NTGCANCNTT
              721 ATNANTGGTT ACNAATAAAA GCAANNAGCA TCANNAATTT TCACAAAATA AAGCATTTTT
              781 TTCACTGCAT NCNANNTTNG NGGNTTGTCC AAACTCATCA ANGNNNNNNN NNCANGTNNG
            841 NNNNACNNNN NNNNNN
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Iida, T., Whole-Genome DNA Methylation Analyses Revealed Epigenetic Instability in Tumorigenic Human iPS Cell-Derived Neural Stem/Progenitor Cells. Stem Cells 35 (5): 1316-1327 (2017). PMID 28142229. [link to RRC of NBRP]
            reference Kojima, K., Selective Ablation of Tumorigenic Cells Following Human Induced Pluripotent Stem Cell-Derived Neural Stem/Progenitor Cell Transplantation in Spinal Cord Injury. Stem Cells Transl. Med. 8 (3): 260-270 (2019). PMID 30485733.
            reference Iwasawa, C., Increased Cytotoxicity of Herpes Simplex Virus Thymidine Kinase Expression in Human Induced Pluripotent Stem Cells. Int. J. Mol. Sci. 20 (4). pii: E810 (2019). PMID 30769780.
            reference Yoshimatsu, S., Evaluating the efficacy of small molecules for neural differentiation of common marmoset ESCs and iPSCs. Neurosci. Res. pii: S0168-0102 (19) 30262-7 (2019). PMID 31586586.
            reference Kwon, S.J., Role of MEK partner-1 in cancer stemness through MEK/ERK pathway in cancerous neural stem cells, expressing EGFRviii. Mol. Cancer 16 (1): 140 (2017). PMID 28830458.

            2024.03.26

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