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pMK278 (mClover-Hygro)

A plasmid for construction of C-terminal mClover tagging donors.

Catalog number RDB13921
Resource name pMK278 (mClover-Hygro)
Alternative name mClover-Hygro, 1000
Clone info. A plasmid for construction of C-terminal mClover tagging donors. Please refer Fig. 4 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use this clone.
Vector backbone pBluescriptII KS- (Plasmid)
Size of vector backbone 3.0 kb
Selectable markers Am^r
Gene/insert name Aequorea victoria GFP cDNA
Depositor|Developer Kanemaki, Masato |
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External Database
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 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.
            Ordering Please visit Information of Request for Distribution.[link] 
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            Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
            Schedule A (GFP-A) [Link]
            Remarks
            Please use Schedule A (GFP-A) additionally.
            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。
            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]
            提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]
            Schedule A(GFP-A, 英語版のみ) [Link]
            備考
            Schedule A(GFP-A, 英語版のみ)が追加で必要です。

            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB13921 pMK278 (mClover-Hygro) 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)
            Featured content Genome Editing (English text)
            Featured content Genome Editing (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 RDB13921_A6B8p1_3.pdf check

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

            Primer: M13_-40
            Region: T7pro,insert5'(mClover(A206K),SV40p(A)
            Sequence file: RDB13921_A6B8e.seq check
            >D03070A05_A6B8_1_M13(-40)_B02_05.ab1
                1 CCGCCGGTAG CGCGCGTATA CGACTCACTA TAGGGCGAAT TGGAGCTCGG ATCCGGTGCA
               61 GGCGCCGCTA GCATGGTGAG CAAGGGCGAG GAGCTGTTCA CCGGGGTGGT GCCCATCCTG
              121 GTCGAGCTGG ACGGCGACGT AAACGGCCAC AAGTTCAGCG TCCGCGGCGA GGGCGAGGGC
              181 GATGCCACCA ACGGCAAGCT GACCCTGAAG TTCATCTGCA CCACCGGCAA GCTGCCCGTG
              241 CCCTGGCCCA CCCTCGTGAC CACCTTCGGC TACGGCGTGG CCTGCTTCAG CCGCTACCCC
              301 GACCACATGA AGCAGCACGA CTTCTTCAAG TCCGCCATGC CCGAAGGCTA CGTCCAGGAG
              361 CGCACCATCT CTTTCAAGGA CGACGGTACC TACAAGACCC GCGCCGAGGT GAAGTTCGAG
              421 GGCGACACCC TGGTGAACCG CATCGAGCTG AAGGGCATCG ACTTCAAGGA GGACGGCAAC
              481 ATCCTGGGGC ACAAGCTGGA GTACAACTTC AACAGCCACA ACGTCTATAT CACGGCCGAC
              541 AAGCAGAAGA ACGGCATCAA GGCTAACTTC AAGATCCGCC ACAACGTTGA GGACGGCAGC
              601 GTGCAGCTCG CCGACCACTA CCAGCAGAAC ACCCCCATCG GCGACGGCCC CGTGCTGCTG
              661 CCCGACAACC ACTACCTGAG CCATCAGTCC AAGCTGAGCA AAGACCCCAA CGAGAAGCGC
              721 GATCACATGG TCCTGCTGGA GTTCGTGACC GCCGCCGGGA TTACACATGG CATGGACGAG
              781 CTGTACAAGT AATCTAGATA ACTGATCATA ATCAGCCATA CCACATTTGT AGAGGTTTTA
              841 CTTGCTTTAA AAAACCTCCC ACACCTCCCC CTGAACCTGA AACATAAAAT GAATGCAATT
              901 GTTGTTGTTA ACTTGTTTAT TGCAGCTTAT AATGGTTACA AATAAGCAAT AGCATCACAA
              961 ATTTCACAAT AAAGCATTTT TTTTCACTGC ATTCTAGTTG TGGTTTTGTC CAAACTCATC
             1021 AATGTATCTT AACGCGTCGA TCATATTCAT AACCTTATAT AACTTCGTAA TAATGTATGC
             1081 TATTACGAGG TTATTAGGTC TGAGAGAGTT AACGTTCCAG CAAGCTTAGG AACCTCGAAC
            1141 TCTCGAGACT
            //
            Primer: PGKpro_F
            Region: PGK pro,insert mid(HygR)
            Sequence file: RDB13921_A6B8f.seq check
            >D03070A05_A6B8_1_PGK-proF_A07_01.ab1
                1 ACTACGCGTT CTCTCTTCTC ATCTCCGGGC CTTTCGACCT GCATCCATCT AGATCTCGAG
               61 CAGCTGAAGC TTACCGCTAG CATGGATAGA TCCGGAAAGC CTGAACTCAC CGCGACGTCT
              121 GTCGAGAAGT TTCTGATCGA AAAGTTCGAC AGCGTCTCCG ACCTGATGCA GCTCTCGGAG
              181 GGCGAAGAAT CTCGTGCTTT CAGCTTCGAT GTAGGAGGGC GTGGATATGT CCTGCGGGTA
              241 AATAGCTGCG CCGATGGTTT CTACAAAGAT CGTTATGTTT ATCGGCACTT TGCATCGGCC
              301 GCGCTCCCGA TTCCGGAAGT GCTTGACATT GGGGAATTCA GCGAGAGCCT GACCTATTGC
              361 ATCTCCCGCC GTGCACAGGG TGTCACGTTG CAAGACCTGC CTGAAACCGA ACTGCCCGCT
              421 GTTCTGCAGC CGGTCGCGGA GGCCATGGAT GCGATCGCTG CGGCCGATCT TAGCCAGACG
              481 AGCGGGTTCG GCCCATTCGG ACCGCAAGGA ATCGGTCAAT ACACTACATG GCGTGATTTC
              541 ATATGCGCGA TTGCTGATCC CCATGTGTAT CACTGGCAAA CTGTGATGGA CGACACCGTC
              601 AGTGCGTCCG TCGCGCAGGC TCTCGATGAG CTGATGCTTT GGGCCGAGGA CTGCCCCGAA
              661 GTCCGGCACC TCGTGCACGC GGATTTCGGC TCCAACAATG TCCTGACGGA CAATGGCCGC
              721 ATAACAGCGG TCATTGACTG GAGCGAGGCG ATGTTCGGGG ATTCCCAATA CGAGGTCGCC
              781 AACATCTTCT TCTGGAGCCG TGGTTGGCTT GTATGGAGCA GCAGACGCGC TACTTCGAGC
              841 GGAGGCATCC GGAGCTTGCA GGATCGCCGC GGCTCCGGGC GTATATGCTC CGCATTGGTC
              901 TTGACCAACT CTATCAGAGC TTGGTTGACG CAATTTCGAT GATGCAGCTT GGGCGCAAGG
              961 GTTCGATGCG ACGCATCGTT CCGATCGGAG TCTGGGACTG TCGGGCGTAC ACAAATCGCC
             1021 GCAGAGCGCG CCGTCTTGGA ACCGAATGCT GTGTAGAGTA CCTCGCGATA GTGAAATCGG
             1081 ACGCTCAGCA CCTGTCGAGC AAGGATTGCC TTAGCTGCGG CCCCTCACGA CTCGCTGAGC
            1141 TGTC
            //
            Primer: PGK_N_rev
            Region: lox P,SV40p(A),insert mid(mClover(A206K)
            Sequence file: RDB13921_A6B8g.seq check
            >D03070A05_A6B8_1_PGK-N-rev_A06_03.ab1
                1 CATGGGCGCT CCCTACCGGT AGATTTCGAG GTCGAGATCC TAAGCTTGGC TGGACGTAAA
               61 CTCCTCTTCA GACCTAATAA CTTCGTATAG CATACATTAT ACGAAGTTAT ATTAAGGGTT
              121 ATTGAATATG ATCGACGCGT TAAGATACAT TGATGAGTTT GGACAAACCA CAACTAGAAT
              181 GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT ATTGCTTTAT TTGTAACCAT
              241 TATAAGCTGC AATAAACAAG TTAACAACAA CAATTGCATT CATTTTATGT TTCAGGTTCA
              301 GGGGGAGGTG TGGGAGGTTT TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA
              361 TTATGATCAG TTATCTAGAT TACTTGTACA GCTCGTCCAT GCCATGTGTA ATCCCGGCGG
              421 CGGTCACGAA CTCCAGCAGG ACCATGTGAT CGCGCTTCTC GTTGGGGTCT TTGCTCAGCT
              481 TGGACTGATG GCTCAGGTAG TGGTTGTCGG GCAGCAGCAC GGGGCCGTCG CCGATGGGGG
              541 TGTTCTGCTG GTAGTGGTCG GCGAGCTGCA CGCTGCCGTC CTCAACGTTG TGGCGGATCT
              601 TGAAGTTAGC CTTGATGCCG TTCTTCTGCT TGTCGGCCGT GATATAGACG TTGTGGCTGT
              661 TGAAGTTGTA CTCCAGCTTG TGCCCCAGGA TGTTGCCGTC CTCCTTGAAG TCGATGCCCT
              721 TCAGCTCGAT GCGGTTCACC AGGGTGTCGC CCTCGAACTT CACCTCGGCG CGGGTCTTGT
              781 AGGTACCGTC GTCCTTGAAA GAGATGGTGC GCTCCTGGAC GTAGCCTTCG GGCATGGCGG
              841 ACTTGAAGAA GTCGTGCTGC TTCATGTGGT CGGGGTAGCG GCTGAAAGCA GCCACGCCGT
              901 AGCCGAAGGT GGTCACGAGG GTGGGCCAGG GCACGGGCAG CTTGCCGGTG GTGCAGATGA
              961 ACTTCAGGGT CAAGCTTGCC GTTTGGTGGC ATCGCCCTCG CCCTCGCCCG CGGACGCTGA
             1021 ACTTTGTGGC CGTTAACGTC GGCCGTCAAG CTCGACCAGA TGGCACCACC CCGGTGAACA
             1081 GCTCTTCGCC ATGCCTCACA ATGCTAGCGC GCATGCACGG GATCCGAGCT CCATTTCGCC
            1141 CCTATTTAGT GAAGATCCGC A
            //
            Primer: M13R
            Region: lox P,mPGK ter,HygR
            Sequence file: RDB13921_A6B8h.seq check
            >D03070A05_A6B8_1_M13R_B01_04.ab1
                1 CTTCGCGCAT TACCCTCACT AAAGGGAACA AAAGCTGGGT ACCGGATCCT AGTGAACCTC
               61 TTCGAGGGAC CTAATAACTT CGTATAGCAT ACATTATACG AAGTTATATT AAGGGTTCCG
              121 GATCTCGACC AGCTTCTGAT GGAATTAGAA CTTGGCAAAA CAATACTGAG AATGAAGTGT
              181 ATGTGGAACA GAGGCTGCTG ATCTCGTTCT TCAGGCTATG AAACTGACAC ATTTGGAAAC
              241 CACAGTACTT AGAACCACAA AGTGGGAATC AAGAGAAAAA CAATGATCCC ACGAGAGATC
              301 TATAGATCTA TAGATCATGA GTGGGAGGAA TGAGCTGGCC CTTAATTTGG TTTTGCTTGT
              361 TTAAATTATG ATATCCAACT ATGAAACATT ATCATAAAGC AATAGTAAAG AGCCTTCAGT
              421 AAAGAGCAGG CATTTATCTA ATCCCACCCC ACCCCCACCC CCGTAGCTCC AATCCTTCCA
              481 TTCAAAATGT AGGTACTCTG TTCTCACCCT TCTTAACAAA GTATGACAGG AAAAACTTCC
              541 ATTTTAGTGG ACATCTTTAT TGTTTAATAG ATCATCAATT TCTGCATCCC GGGGATCTGA
              601 TATCATCGAT GCATGGGGTC GTGCGCTCCT TTCGGTCGGG CGCTGCGGGT CGTGGGGCGG
              661 GCGGCTAGCC TATTCCTTTG CCCTCGGACG AGTGCTGGGG CGTCGGTTTC CACTATCGGC
              721 GAGTACTTCT ACACAGCCAT CGGTCCAGAC GGCCGCGCTT CTGCGGGCGA TTTGTGTACG
              781 CCCCGACAGT CCCGGCTCCG GATCGGGACG ATTGCGTCGC ATCGACCCTG CGCCCAAGCT
              841 GCATCATCGA AAATTGCCCG TCAACCAAAG CTCTGATAGA GTTTGGTCAA GACCAATGGC
              901 GGGAGCATAA TACCGCCCGG GGAGCCCGCG GGCGAATCCT GCAAGGCTCC GGGATGCCTC
              961 CGGCTCGAAA GTAAGCGCGT CTGGCTGGCT CCATACAAGG CCAACCACGG GCCTTCCAGA
             1021 AAGAAGATGT TTGGGGACCC TCGTATTGGT GAAAATCTCC CGAACAGTCG GCCCTCCGCC
             1081 TCCAGTTCAA TGAACCGCGC TGTTTAATGC GGGGCCTATT TGTTCCGACT CAGGGGACAT
            1141 ATTGTGTT
            //

            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.

            2022.02.09

            GNP_filter3_RDBDEP_html_220112.pl