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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 |
Other clones in our bank
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 for distribution 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:
          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 contains CRISPR/Cas9 technologies and is not provided to for-profit organization or for for-profit research by non-profit organizations.
          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CRISPR/Cas9 technologyを用いたゲノム編集バイオリソースです。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。

          Catalog # Resource name Shipping form Fee (non-profit org.)
          RDB13917 AAVS1 T2 CRISPR in pX330 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 AAVS1 T2 CRISPR in pX330 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13917).

          Reference section:

          Natsume, T., Kiyomitsu, T., Saga, Y., Kanemaki, M.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)

          References and tips

          Featured content

          Featured content Auxin Inducible Degron (AID) System (English text)
          Featured content Auxin Inducible Degron (AID) System (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 RDB13917_A6B8p1-4.pdf check

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

          Primer: SV40pA3'element Reverse (Pr0108)
          Region: U6 pro, gRNA, gRNA scaffold
          Sequence file: RDB13917_A6B8c.seq check
          >13917_13917_B2JjF_U6pro_F3_E02_14_ABI24.ab1
              1 NNNNNNNNTN NNANNGGNNN ATNNTATGCT TACCGTAACT TGAAAGTATT TCGATTTCTT
             61 GGCTTTATAT ATCTTGTGGA AAGGACGAAA CACCGGGGCC ACTAGGGACA GGATGTTTTA
            121 GAGCTAGAAA TAGCAAGTTA AAATAAGGCT AGTCCGTTAT CAACTTGAAA AAGTGGCACC
            181 GAGTCGGTGC TTTTTTGTTT TAGAGCTAGA AATAGCAAGT TAAAATAAGG CTAGTCCGTT
            241 TTTAGCGCGT GCGCCAATTC TGCAGACAAA TGGCTCTAGA GGTACCCGTT ACATAACTTA
            301 CGGTAAATGG CCCGCCTGGC TGACCGCCCA ACGACCCCCG CCCATTGACG TCAATAGTAA
            361 CGCCAATAGG GACTTTCCAT TGACGTCAAT GGGTGGAGTA TTTACGGTAA ACTGCCCACT
            421 TGGCAGTACA TCAAGTGTAT CATATGCCAA GTACGCCCCC TATTGACGTC AATGACGGTA
            481 AATGGCCCGC CTGGCATTGT GCCCAGTACA TGACCTTATG GGACTTTCCT ACTTGGCAGT
            541 ACATCTACGT ATTAGTCATC GCTATTACCA TGGTCGAGGT GAGCCCCACG TTCTGCTTCA
            601 CTCTCCCCAT CTCCCCCCCC TCCCCACCCC CAATTTTGTA TTTATTTATT TTTTAATTAT
            661 TTTGTGCAGC GATGGGGGCG GGGGGGGGGG GGGGGNNCGC GCCAGGCGGG GCGGGGCGGG
            721 NCNNGGGGCG GGGCGGGNCN AGGCGNANAG GTGCGGCGGC ANCCAATCAN AGCGGCGCGC
            781 TCCGAAAGTT NCCTTTNATG GNNAGGNGNN GGCGGCGGCN NCCCTATAAA AAGCNNNNCG
            841 CGCGGNNGGG CGGGANNCGC NGCGCNCTGC NTNNGCCCNG NNCNNNNTCC GNNNNNNNCN
            901 TCGNNNNGCC CNNCCCNNGG CTCTNNNNGA NNNCGTNNNT NCNNNNNGNN ANNNGNNNGG
          961 NNNNNNNNNN
          //
          Primer: RDB13535#1 (Pr0302)
          Region: CBh pro, 3xFLAG, SV40 NLS, Cas9 5'
          Sequence file: RDB13917_A6B8d.seq check
          >13917_13917_B2JjF_Cas9-N_R1_E02_05_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNGN NNNNNNNNNT GTACTCGTCG GTGANCACGG CCCAGCCCAC
             61 AGAGTTGGTG CCGATGTCCA GGCCGATGCT GTACTTCTTG TCGGCTGCTG GGACTCCGTG
            121 GATACCGACC TTCCGCTTCT TCTTTGGGGC CATCTTATCG TCATCGTCTT TGTAATCAAT
            181 ATCATGATCC TTGTAGTCTC CGTCGTGGTC CTTATAGTCC ATGGTGGCAC CGGTCCAACC
            241 TGAAAAAAAG TGATTTCAGG CAGGTGCTCC AGGTAATTAA ACATTAATAC CCCACCAACC
            301 AACCATCCCT TAAACCCTTA CCTCTTGCTC AGCTAATTAC AGCCCGGAGG AGAAGGGCCG
            361 TCCCGCCCGC TCACCTGTGG GAGTAACGCG GTCAGTCAGA GCCGGGGCGG GCGGCGCGAG
          421 GCGN
          //
          Primer: bGH_rev3 (Pr0885)
          Region: BGH pA,nucleoplasmin NLS,Cas9
          Sequence file: RDB13917_A6B8e.seq check
          >13917_13917_B2JjF_bGH_rev3_A01_01_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNGGG CNNNCAACAG ATGGCTGGCA ACTAGAAGGC
             61 ACAGTCGAGG CTGATCAGCG AGCTCTAGGA ATTCTTACTT TTTCTTTTTT GCCTGGCCGG
            121 CCTTTTTCGT GGCCGCCGGC CTTTTGTCGC CTCCCAGCTG AGACAGGTCG ATCCGTGTCT
            181 CGTACAGGCC GGTGATGCTC TGGTGGATCA GGGTGGCGTC CAGCACCTCT TTGGTGCTGG
            241 TGTACCTCTT CCGGTCGATG GTGGTGTCAA AGTACTTGAA GGCGGCAGGG GCTCCCAGAT
            301 TGGTCAGGGT AAACAGGTGG ATGATATTCT CGGCCTGCTC TCTGATGGGC TTATCCCGGT
            361 GCTTGTTGTA GGCGGACAGC ACTTTGTCCA GATTAGCGTC GGCCAGGATC ACTCTCTTGG
            421 AGAACTCGCT GATCTGCTCG ATGATCTCGT CCAGGTAGTG CTTGTGCTGT TCCACAAACA
            481 GCTGTTTCTG CTCATTATCC TCGGGGGAGC CCTTCAGCTT CTCATAGTGG CTGGCCAGGT
            541 ACAGGAAGTT CACATATTTG GAGGGCAGGG CCAGTTCGTT TCCCTTCTGC AGTTCGCCGG
            601 CAGAGGCCAG CATTCTCTTC CGGCCGTTTT CCAGCTCGAA CAGGGAGTAC TTAGGCAGCT
            661 TGATGATCAG GTCCTTTTTC ACTTCTTTGT AGCCCTTGGC TTCCAGAAAG TCGATGGGAT
            721 TCTTCTCGAA GCTGCTTCTT TCCATGATGG TGATCCCCAG CAGCTCTTTC ACACTCTTCA
            781 GTTTCTTGGA CTTGCCCTTT TCCACTTTGG CCACCACCAG CACAGAATAN GCCACGGNGG
            841 GGCTGTNNAA GCCGCCGTAC TTCTTANGGT CCCAGTCCTT CTTTCTGGCG ATCAGCTTAT
            901 CNCTGTNNCN CTNNNNNANG ANANNCTCTT NGCTGNNNCC NNCTNNNTGC NNCNCNGNCN
          961 TTTTCACNNN ANTNNNNNNN GGNNNNGNNN NNNNNNTNNN NNNNNNN
          //

          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.

          2023.05.22

          GNP_filter3_RDBDEP_html_230512.pl