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AAVS1 T2 CRISPR in pX330

A CRISPR/Cas plasmid for making DNA dounle-strand break at the human AAVS1 locus.

Catalog number RDB13917
Resource name AAVS1 T2 CRISPR in pX330
Clone info. A CRISPR/Cas plasmid for making DNA dounle-strand break at the human AAVS1 locus.
Vector backbone pX330-U6-Chimeric_BB-CBh-hSpCas9 (Plasmid)
Size of vector backbone 4.3 kb
Selectable markers Am^r
Gene/insert name S. pyogenes. Cas9 cDNA
Depositor|Developer Kanemaki, Masato |
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External Database
S. pyogenes. Cas9

          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 the following literature designated by the DEPOSITOR is requested (Natsume, T., Cell Reports, 15: 210-218, 2016). This DNA clone is used for academic research purpose only. This is not to be used on a commercial basis. For use of this DNA clone except for internal academic research, the RECIPIENT must contact a NIG license representative at chizai@nig.ac.jp.
          Additional terms and conditions for distribution Regarding resources containing CAG promoter: In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation for the CAG promoter (Niwa, H., Yamamura, K., Miyazaki, J., Gene 108 : 193-200, 1991) and an acknowledgment to Dr. Jun-ich Miyazaki of the Osaka University are requested.
          Ordering Please visit Information of Request for Distribution.[link] 
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          Exclusive MTA (For the DNA materials containing CRISPR/Cas9 technologies and for not-for-profit academic purpose) [Word]
          Remarks
          The BIOLOGICAL RESOURCE is not provided to for-profit organization or for for-profit research by non-profit organizations. Please refer the Exclusive MTA (For the DNA materials containing CRISPR/Cas9 technologies and for not-for-profit academic purpose).
          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を引用する (Natsume, T., Cell Reports, 15: 210-218, 2016)。学術研究にのみ限定、商業営利目的利用を禁ずる。学術内部利用以外の利用の際は、国立遺伝学研究所 (chizai@nig.ac.jp) に相談すること。
          付加的提供条件 CAGプロモータを含むリソースについて: 利用者は、研究成果の公表にあたってCAGプロモータの文献 (Niwa, H., Yamamura, K., Miyazaki, J., Gene 108 : 193-200, 1991)を引用し、大阪大学 宮崎純一博士への謝辞の表明を必要とする。
          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。
          提供依頼書 [Word]
          専用MTA(CRISPR/Cas9内包遺伝子材料専用 非営利学術目的)をお使いください [Word]
          備考
          本件リソースは営利機関および非営利機関による営利目的研究には提供いたしません。専用MTA(CRISPR/Cas9内包遺伝子材料専用 非営利学術目的)をご参照ください。

          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB13917 AAVS1 T2 CRISPR in pX330 DNA solution

          Ordering Information [in Japanese] [in English]
          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 Natsume, T., Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15 (1): 210-218 (2016). PMID 27052166.

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

          Featured content

          Featured content Auxin Inducible Degron (AID) System (English text)
          Featured content Auxin Inducible Degron (AID) System (Japanese 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 RDB13917_A6B8p1-3.pdf check

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

          Primer: SV40pA3'element Reverse
          Region: U6 pro, gRNA, gRNA scaffold
          Sequence file: RDB13917_A6B8c.seq check
          >D03070A01_A6B8_1_SV40pA3'element_Reverse_G12_21.ab1
              1 GGCGCAGGAG GCGGACCTAT GGAAACGCCA GCAACGCGGC CTTTTTACGG TTCCTGGCCT
             61 TTTGCTGGCC TTTTGCTCAC ATGTGAGGGC CTATTTCCCA TGATTCCTTC ATATTTGCAT
            121 ATACGATACA AGGCTGTTAG AGAGATAATT GGAATTAATT TGACTGTAAA CACAAAGATA
            181 TTAGTACAAA ATACGTGACG TAGAAAGTAA TAATTTCTTG GGTAGTTTGC AGTTTTAAAA
            241 TTATGTTTTA AAATGGACTA TCATATGCTT ACCGTAACTT GAAAGTATTT CGATTTCTTG
            301 GCTTTATATA TCTTGTGGAA AGGACGAAAC ACCGGGGCCA CTAGGGACAG GATGTTTTAG
            361 AGCTAGAAAT AGCAAGTTAA AATAAGGCTA GTCCGTTATC AACTTGAAAA AGTGGCACCG
            421 AGTCGGTGCT TTTTTGTTTT AGAGCTAGAA ATAGCAAGTT AAAAATAAAG GCTAGTCCGT
            481 TTTAGCGCGT GCGCCAATTC TGCAGAACAA ATGGCTCTAG AAGGTACCCG TTACCATAAC
            541 TTTACGGTAA AATGGGCCCG CCTGGCTGAC CGCCCAACGA CCCCCCGCCC CATTGACGTC
            601 AATAGTAACG CCCAATAGGG ACCTTTCCAT TGAACGTCAA GGGGTGAGGA ATTACGCGTA
            661 AAACTGCCAC TGGGCGGACA TACAGTGTGA TCTATTGCCA GGTACGCCCC CTATGACGTC
            721 ATGACGTTAA TGGCCCCTGC TTTGTCCATA CATGACCTAT GGAACTTCTC ACTGGCGAAC
            781 TCAACGATTA GACCGCCGAT ACATGGCGAG TTGATCAAAG TTTTTGTTTT TTATTTCTCA
          841 GGATGCGCGC GCGTTGCGAA CCCCCAATTT GGAAATATA
          //
          Primer: RDB13535#1
          Region: CBh pro, 3xFLAG, SV40 NLS, Cas9 5'
          Sequence file: RDB13917_A6B8d.seq check
          >D03070A01_A6B8_1_RDB13535#1_H12_24.ab1
              1 TTGACATGCC TGAATCACTT TTTTTCAGGT TGGACCGGTG CCACCATGGA CTATAAGGAC
             61 CACGACGGAG ACTACAAGGA TCATGATATT GATTACAAAG ACGATGACGA TAAGATGGCC
            121 CCAAAGAAGA AGCGGAAGGT CGGTATCCAC GGAGTCCCAG CAGCCGACAA GAAGTACAGC
            181 ATCGGCCTGG ACATCGGCAC CAACTCTGTG GGCTGGGCCG TGATCACCGA CGAGTACAAG
            241 GTGCCCAGCA AGAAATTCAA GGTGCTGGGC AACACCGACC GGCACAGCAT CAAGAAGAAC
            301 CTGATCGGAG CCCTGCTGTT CGACAGCGGC GAAACAGCCG AGGCCACCCG GCTGAAGAGA
            361 ACCGCCAGAA GAAGATACAC CAGACGGAAG AACCGGATCT GCTATCTGCA AGAGATCTTC
            421 AGCAACGAGA TGGCCAAGGT GGACGACAGC TTCTTCCACA GACTGGAAGA GTCCTTCCTG
            481 GTGGAAGAGG ATAAGAAGCA CGAGCGGCAC CCCATCTTCG GCAACATCGT GGACGAGGTG
            541 GCCTACCACG AGAAGTACCC CACCATCTAC CACCTGAGAA AGAAACTGGT GGACAGCACC
            601 GACAAGGCCG ACCTGCGGCT GATCTATCTG GCCCTGGCCC ACATGATCAA GTTCCGGGGC
            661 CACTTCCTGA TCGAGGGCGA CCTGAACCCC GACAACAGCG ACGTGGACAA GCTGTTCATC
            721 CAGCTGGTGC AGACCTACAA CCAGCTGTTC GAGGAAAACC CCATCAACGC CAGCGGCGTG
            781 GACGCCAAGG CCATCCTGTC TGCCAGACTG AGCAAGAGCA GACGGCTGGA AAATCTGATC
            841 GCCCAGCTGC CCGGCGAGAA GAAGAATGGC CTGTTCGGAA ACCTGATTGC CCTGAGCCTG
            901 GGCCTGACCC CCAACTTCAA GAGCAACTTC GACCTGGCCC GAGGATGCCA AACTGCAGCT
            961 GAGCAAGGAC ACCTACGACG ACGACCTGGA CAAACCTGCT GGCCAAGATC GGCGACCAGT
           1021 ACGCCCGACC TTGTTTCTGG CCGCCAAGAA CCTTGTCCGG ACGCCATCAT GCCTGACCGG
           1081 ACATCCTGGA GAGTTGACAC GAGATCAACT AAGCACCCCT GGACGGCCTC TTATGGAATT
          1141 CCAAG
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          original Natsume, T., Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15 (1): 210-218 (2016). PMID 27052166.
          reference Natsume, T., Acute inactivation of the replicative helicase in human cells triggers MCM8-9-dependent DNA synthesis. Genes Dev. (2017). PMID 28487407.

          2022.02.09

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