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CTCF-mAC donor (Hygro) (#RDB18354)

Donor plasmid to generate CTCF-mAID degron cell lines with mClover as a marker, hygromycin resistance.

Clone info. Donor plasmid to generate CTCF-mAID degron cell lines with mClover as a marker, hygromycin resistance, see Fig. 4E of Yesbolatova, A. et al., Nat. Commun. 11: 5701, 2020.
Vector backbone pBluescriptII KS- (plasmid)
Selectable markers Ampicillin (E. coli), Hygromycin (mammalian cell).
Gene/insert name human CTCF genomic Arabidopsis thaliana IAA17 cDNA Aequorea victoria GFP (mClover) cDNA
Depositor|Developer Kanemaki, Masato |
 
Sequence (full) RDB18354zzk01.seq provided by the depositor

Distribution information

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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 is requested (Yesbolatova, A. et al., Nat. Commun. 11: 5701 (2020)).
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提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
手続きの概要は、「提供申込みについて[link]」をご覧ください。
MTAに書く使用条件 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 (Yesbolatova, A. et al., Nat. Commun. 11: 5701 (2020))。

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RDB18354 CTCF-mAC donor (Hygro) 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 CTCF-mAC donor (Hygro) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB18354).

Reference section:

Yesbolatova, A., Saito, Y., Kitamoto, N., Makino-Itou, H., Ajima, R., Nakano, R., Nakaoka, H., Fukui, K., Gamo, K., Tominari, Y., Takeuchi, H., Saga, Y., Hayashi, K., Kanemaki, M.T., The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Nat. Commun. 11: 5701 (2020). PMID 33177522. [link to RRC of NBRP]

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

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 RDB18354_B2Hap1-1.pdf check

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

Primer: EGFP-M_R (Pr0743)
Region: insert mid(mClover 5',mAID 3')
Sequence file: RDB18354_B2Haa.seq check
>D07105C4_B2Ha_3_EGFP-M_R_D01_10_ABI24.ab1
    1 NNNNNNNNNN NNANNTNNTG NNGGNNCTNN NNNTAGCCTT GATGCCGTTC TTCTGCTTGT
   61 CGGNNNNNNN NTANACGTTG TGGCTGTTGA AGTTGTACTC CAGCTTGTGC CCCAGGATGT
  121 TGCCGTCCTC CTTGAAGTCG ATGCCCTTCA GCTCGATGCG GTTCACCAGG GTGTCGCCCT
  181 CGAACTTCAC CTCGGCGCGG GTCTTGTAGG TACCGTCGTC CTTGAAAGAG ATGGTGCGCT
  241 CCTGGACGTA GCCTTCGGGC ATGGCGGACT TGAAGAAGTC GTGCTGCTTC ATGTGGTCGG
  301 GGTAGCGGCT GAAGCAGGCC ACGCCGTAGC CGAAGGTGGT CACGAGGGTG GGCCAGGGCA
  361 CGGGCAGCTT GCCGGTGGTG CAGATGAACT TCAGGGTCAG CTTGCCGTTG GTGGCATCGC
  421 CCTCGCCCTC GCCGCGGACG CTGAACTTGT GGCCGTTTAC GTCGCCGTCC AGCTCGACCA
  481 GGATGGGCAC CACCCCGGTG AACAGCTCCT CGCCCTTGCT CACCATGCTA GCTTTATACA
  541 TCCTCAAATC GATTTTCCTC AAGTACGGTG CTCCGTCCAT TGATACCTTC ACGAACGCCG
  601 CCGCCTCCGG GCCACCGCTT GATTTTTGGC AGGAAACCAT CACGTTCTTC CGGTATGATC
  661 TCACCGGTGG CCATCCCACA ACTTGTGCCT TGGCCGGAGG TTTGGCTGGA TCTTTANGAC
  721 AAGCACTCTT CTCCTTGGCG CCTGCACCGG ATCCCCGGTC CATCATGCTG ANGATCATCT
  781 CGGGCGTGAN GTCTCCGTTG GGGGCATCTG TGGCAGCTGG CTGGGCCTCC TCTTCCTCTC
  841 NCTCTGCNGG CTCAGCATCT GGCTCTTTTT TTTACTTCAA CTATAATGNT CTCNATTGCA
  901 CCNNTATTCT GGNCTTCAAC CNGNANTGAT AGCTGTTGCT GGNAAANAAN ANCNCAGANA
961 GAACAAATGG NNCATNNAAG CCCNNNGAAT CANNNNNNNN NTTNNNNNNN NNN
//
Primer: mPGKter_F (Pr0894)
Region: insert 3'(loxP.2,CTCF homology arm.2)
Sequence file: RDB18354_B2Hab.seq check
>D07105C4_B2Ha_3_mPGKter_F_B01_04_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNT GTTTTNCNNA GTTCTAATTC NNTCAGAAGC TGGTCGAGAT
   61 CCGGAACCCT NNNNNNNNNC TTCGTATAAT GTATGCTATA CGAAGTTATT AGGTCCCTCG
  121 AAGAGGTTCA CTAGGATCCG CGTCGCCAGG ACTTCTCTGG GCTGTGTTTA AACGGCCCGC
  181 ATCTTAATTT TTCTCCCTTC TTTCTTTTTT TGGCTTTGGG AAAAGCATCA TTTTACCAAA
  241 CATACCGAGA ACGAAAACTT CAAGGATGAT GTTAGAAAAA AATGTGATTT AACTAGAACT
  301 TGCTGTCTGA TGTTAGCAAA TCATGGAATG TTCTGAGTCC CTGAGGGTTT ACTGTGAAGT
  361 GCTGAGGACA GTGTTGACAA CTAACTCGTT TTCCTAGATG GAAACGGAGA CATTGACCCC
  421 TCCCTCCATG TGGTAAACCA CTCCAGAATG GCCACCAGGC TTCCCAGAGT TCTATGGTCT
  481 TCTTCCCAAG AGAGTTTTTA ATTGTAAATG CATACTTGGG AAGGACTTAG AGTTTTAAAC
  541 TGTTTTTTGC TTTTGCTTTT CCCTGACTCC CTTTGCTTGG AGGGTACCCA GCTTTTGTTC
  601 CCTTTAGTGA GGGTTAATTG CGCGCTTGGC GTAATCATGG TCATAGCTGT TTCCTGTGTG
  661 AAATTGTTAT CCGCTCACAA TTCCACACAA CATACGAGCC GGGAGCATAA AGTGTAAAGC
  721 CTGGNGTGCC TAATGAGTGA GCTAACTCAC ATTAATTGCG TTGCGCTCAC TGCCCGCTTT
  781 CCAGTCGGGA AACCTGTCGT GCCAGCTGCA TTAATGAATC GGCCAACGCG CGGGGANAGG
  841 CGGTTTGCGT ATTGGGCGCT CTTCCGCTTC CTCGCTCACT GANTCGCTGC GCTCGGNCGT
  901 TCGGCTGCNG CNANCGGTAT CAGCTCACTC NAAGGCGGNA ATACNNNTAT NCNCAGNNTC
961 NNNGGNNNAC NCNNNAAGNN NNTGNNANCN AANNNNCANC AAANNNNNNN NNN
//
Primer: PGK_N_rev (Pr0243)
Region: insert mid(SV40 pA,mClover 3')
Sequence file: RDB18354_B2Hac.seq check
>D07105C4_B2Ha_3_PGK_N_rev_E01_13_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NGNANANTTC GAGGTCGAGN NNCCTAAGCT TGGCTGGACG
   61 TAAACTCCTN NNCNNACCTA ATAACTTCGT ATAGCATACA TTATACGAAG TNNNNNTAAG
  121 GGTTATTGAA TATGATCGAC GCGTTAAGAT ACATTGATGA GTTTGGACAA ACCACAACTA
  181 GAATGCAGTG AAAAAAATGC TTTATTTGTG AAATTTGTGA TGCTATTGCT TTATTTGTAA
  241 CCATTATAAG CTGCAATAAA CAAGTTAACA ACAACAATTG CATTCATTTT ATGTTTCAGG
  301 TTCAGGGGGA GGTGTGGGAG GTTTTTTAAA GCAAGTAAAA CCTCTACAAA TGTGGTATGG
  361 CTGATTATGA TCAGTTATCT AGATTACTTG TACAGCTCGT CCATGCCATG TGTAATCCCG
  421 GCGGCGGTCA CGAACTCCAG CAGGACCATG TGATCGCGCT TCTCGTTGGG GTCTTTGCTC
  481 AGCTTGGACT GATGGCTCAG GTAGTGGTTG TCGGGCAGCA GCACGGGGCC GTCGCCGATG
  541 GGGGTGTTCT GCTGGTAGTG GTCGGCGAGC TGCACGCTGC CGTCCTCAAC GTTGTGGCGG
  601 ATCTTGAAGT TAGCCTTGAT GCCGTTCTTC TGCTTGTCGG CCGTGATATA GACGTTGTGG
  661 CTGTTGAAGT TGTACTCCAG CTTGTGCCCC AGGATGTTGC CGTCCTCCTT GAAGTCGATG
  721 CCCTTCAGCT CGATGCGGTT CACCAGGGTG TCGCCCTCGA ACTTCACCTC GGCGCGGGTC
  781 TTGTANGTAC CGTCGTCCTT GAAAGAGATG GTGCGCTCCT GGACGTANCC TTCGGGCATG
  841 GCGGANTTGA ANAANTCGTG CTGCTTCATG TGGTCGGGGT ANCGGCTGAA GCNNNCCACG
  901 CCGTANCCGA NNNNGTCACN ANGGTGGGNC AGGGNACGGG CAGCTTGNCN GTGGTGNNNA
961 NGANNTTCNN GNNCANCNTG NCNNNNGNNN NNTCNNNNNG CCNNNNNNNN NN
//
Primer: PGKpro_F (Pr0249)
Region: insert mid(HygR)
Sequence file: RDB18354_B2Had.seq check
>D07105C4_B2Ha_3_PGKpro_F_C01_07_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNTCTCCNGG CCTTTCGACC TGCATCCNTC TAGATCTCGA
   61 GCAGCTGANN NNTNNNNCTA GCATGGATAG ATCCGGAAAG CCTGAACTCA CCGCGACGTC
  121 TGTCGAGAAG TTTCTGATCG AAAAGTTCGA CAGCGTCTCC GACCTGATGC AGCTCTCGGA
  181 GGGCGAAGAA TCTCGTGCTT TCAGCTTCGA TGTAGGAGGG CGTGGATATG TCCTGCGGGT
  241 AAATAGCTGC GCCGATGGTT TCTACAAAGA TCGTTATGTT TATCGGCACT TTGCATCGGC
  301 CGCGCTCCCG ATTCCGGAAG TGCTTGACAT TGGGGAATTC AGCGAGAGCC TGACCTATTG
  361 CATCTCCCGC CGTGCACAGG GTGTCACGTT GCAAGACCTG CCTGAAACCG AACTGCCCGC
  421 TGTTCTGCAG CCGGTCGCGG AGGCCATGGA TGCGATCGCT GCGGCCGATC TTAGCCAGAC
  481 GAGCGGGTTC GGCCCATTCG GACCGCAAGG AATCGGTCAA TACACTACAT GGCGTGATTT
  541 CATATGCGCG ATTGCTGATC CCCATGTGTA TCACTGGCAA ACTGTGATGG ACGACACCGT
  601 CAGTGCGTCC GTCGCGCAGG CTCTCGATGA GCTGATGCTT TGGGCCGAGG ACTGCCCCGA
  661 AGTCCGGCAC CTCGTGCACG CGGATTTCGG CTCCAACAAT GTCCTGACGG ACAATGGCCG
  721 CATAACAGCG GTCATTGACT GGAGCGAGGC GATGTTCGGG GATTCCCAAT ACGANGTCGC
  781 CAACATCTTC TTCTGGANGC CGTGGTTGGC TTGTATGGAG CAGCAGACGC GCTACTTCNA
  841 GCGGANGCAT CCGGANCTTG CANGATCGCC GCGGCTCCNG GCGTATATGC TCCGCANTGG
  901 TCTTGANCNN CTCTATCAGA NCNNNGNTNG ANGGCNATTT CGATGANGCN NCTTGNNNNC
961 NNGNNCGATG CNANNCNNNC NNNNNNTNNN GNANNCNNGN NNNGNNGGGN NNNNNNNNN
//
Primer: pUC-CAPbs_up_F2 (Pr0739)
Region: insert 3'(CTCF homology arm.2, loxP.2,mPGK ter)
Sequence file: RDB18354_B2Hae.seq check
>D07105C4_B2Ha_3_pUC-CAPbs_up_F2_G01_19_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNCTTTNNNC TTTATGCTCC CGGCTCGTAT GTTGTGTGGA
   61 ATTGTGAGCG GNNNNNAATT TCACACAGGA AACAGCTATG ACCATGATTA CGCCNAGCGC
  121 GCAATTAACC CTCACTAAAG GGAACAAAAG CTGGGTACCC TCCAAGCAAA GGGAGTCAGG
  181 GAAAAGCAAA AGCAAAAAAC AGTTTAAAAC TCTAAGTCCT TCCCAAGTAT GCATTTACAA
  241 TTAAAAACTC TCTTGGGAAG AAGACCATAG AACTCTGGGA AGCCTGGTGG CCATTCTGGA
  301 GTGGTTTACC ACATGGAGGG AGGGGTCAAT GTCTCCGTTT CCATCTAGGA AAACGAGTTA
  361 GTTGTCAACA CTGTCCTCAG CACTTCACAG TAAACCCTCA GGGACTCAGA ACATTCCATG
  421 ATTTGCTAAC ATCAGACAGC AAGTTCTAGT TAAATCACAT TTTTTTCTAA CATCATCCTT
  481 GAAGTTTTCG TTCTCGGTAT GTTTGGTAAA ATGATGCTTT TCCCAAAGCC AAAAAAAGAA
  541 AGAAGGGAGA AAAATTAAGA TGCGGGCCGT TTAAACACAG CCCAGAGAAG TCCTGGCGAC
  601 GCGGATCCTA GTGAACCTCT TCGAGGGACC TAATAACTTC GTATAGCATA CATTATACGA
  661 AGTTATATTA AGGGTTCCGG ATCTCGACCA GCTTCTGATG GAATTAGAAC TTGGCAAAAC
  721 AATACTGAGA ATGAAGTGTA TGTGGAACAG ANGCTGCTGA TCTCGTTCTT CANGCTATGA
  781 AACTGACACA TTTGGAAACC ACAGTACTTA GAACCACAAA GTGGGAATCA AGAGAAAAAC
  841 AATGATCCCA CGAGAGATCT ATAGATCTAT ANATCATGAN TGGGANGAAT GAGCTGGNCC
  901 TTNATTTGGT TTTNNTTGNT TAAATTATGA TATNCNACTA TGAAACATTA TCATAAAGNN
  961 NTAGTAAAGA NCCTNCNNTN AANANNANGN ATTTNNCTNA NTCCNNCCCN NNCCCNNNNN
1021 NCNNNNNNNN NN
//
Primer: pUC-M13_-100 (Pr0310)
Region: insert 5'(T7 pro,CTCF homology arm.1,mAID 5')
Sequence file: RDB18354_B2Haf.seq check
>D07105C4_B2Ha_3_pUC-M13_-100_F01_16_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NGGNANGCNN GGGTTTTCCC NGTCNCGACG TTGTAAAACG
   61 ACGGCCAGTG AGCGCGCGTA ATACGACTCA CTATAGGGCG AATTGGAGCT CTGAGGCAGG
  121 AGAATTGCTT TAACCCGGGG GGCAGAGGTT GCAGTGAGCT GAGATCATGC CCGTGCACTC
  181 CAGCCTGGTC ATAGAGCAAG ACTCCGTCTC AAAAAATAAA ATAAAAAATA AATTGACTGT
  241 CTCTGGACCG CTATCTAATA AGGCTGTCCT GCATTGCTGA CATCCCGTTC GCTGTCAGTC
  301 TAAAAGACCC TTGTGATTCT TGGGGCTTTA ATGGACCATT TGTTCTGTCT GTGCTCTTCT
  361 TTGCCAGCAA CAGCTATCAT TCAGGTTGAA GACCAGAATA CAGGTGCAAT TGAGAACATT
  421 ATAGTTGAAG TAAAAAAAGA GCCAGATGCT GAGCCCGCAG AGGGAGAGGA AGAGGAGGCC
  481 CAGCCAGCTG CCACAGATGC CCCCAACGGA GACCTCACGC CCGAGATGAT CCTCAGCATG
  541 ATGGACCGGG GATCCGGTGC AGGCGCCAAG GAGAAGAGTG CTTGTCCTAA AGATCCAGCC
  601 AAACCTCCGG CCAAGGCACA AGTTGTGGGA TGGCCACCGG TGAGATCATA CCGGAAGAAC
  661 GTGATGGTTT CCTGCCAAAA ATCAAGCGGT GGCCCGGANG CGGCGGCGTT CGTGAAGGTA
  721 TCAATGGACG GAGCACCGTA CTTGANGAAA ATCGATTTGA GGANGTATAA AGCTAGCATG
  781 GTGAGCAAGG NCGAGGANCT GTTCACCGGG GTGGTGCCCA TCCTGGTCGA NCTGGACNGC
  841 GACGTAAACG GCCACAAGTT CAGCGTCCGC GGCGAGGGCG ANGGCGANGN CNCCNACGGC
  901 AAGCTGANCC NGAAGTTCAT CTGCACCACC NGNNAGCTGC CCNNNCCCNN NNCCNNNCNN
961 CNNGNNCANC NTNCNNNNAN NNNNNNNCNN GNTTCANNCN GCTACCCNNN NNNNNNN
//
Primer: YFP_R (Pr0127)
Region: insert 5'(mClover 5',mAID,CTCF homology arm.1)
Sequence file: RDB18354_B2Hag.seq check
>D07105C4_B2Ha_3_YFP_R_A01_01_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNCN GNNGCTNANT TGTGGCCGTT TACGTCGCCG TCCAGCTCGA
   61 CCAGGANGGN NACCACCCCG GTGAACAGCT CCTCGCCCTT GCTCACCATG CTAGCTTTAT
  121 ACATCCTCAA ATCGATTTTC CTCAAGTACG GTGCTCCGTC CATTGATACC TTCACGAACG
  181 CCGCCGCCTC CGGGCCACCG CTTGATTTTT GGCAGGAAAC CATCACGTTC TTCCGGTATG
  241 ATCTCACCGG TGGCCATCCC ACAACTTGTG CCTTGGCCGG AGGTTTGGCT GGATCTTTAG
  301 GACAAGCACT CTTCTCCTTG GCGCCTGCAC CGGATCCCCG GTCCATCATG CTGAGGATCA
  361 TCTCGGGCGT GAGGTCTCCG TTGGGGGCAT CTGTGGCAGC TGGCTGGGCC TCCTCTTCCT
  421 CTCCCTCTGC GGGCTCAGCA TCTGGCTCTT TTTTTACTTC AACTATAATG TTCTCAATTG
  481 CACCTGTATT CTGGTCTTCA ACCTGAATGA TAGCTGTTGC TGGCAAAGAA GAGCACAGAC
  541 AGAACAAATG GTCCATTAAA GCCCCAAGAA TCACAAGGGT CTTTTAGACT GACAGCGAAC
  601 GGGATGTCAG CAATGCAGGA CAGCCTTATT AGATAGCGGT CCAGAGACAG TCAATTTATT
  661 TTTTATTTTA TTTTTTGAGA CGGAGTCTTG CTCTATGACC AGGCTGGAGT GCACGGGCAT
  721 GATCTCAGCT CACTGCAACC TCTGCCCCCC GGGTTAAAGC AATTCTCCTG CCTCAGANCT
  781 CCAATTCGCC CTATAGTGAG TCGTANTACG CGCGCTCACT GGCCGTCGTT TTACAACGTC
  841 GTGACTGGGA AAACCCTGGC GTTNCCCAAC TTAATCGCCT TGCAGCACAT CCCCCTTTCG
  901 CCNGCTGGCG TAATANCNAA NANGNCCGCA CCGATCGCCC NNCCNNNCAN TTGCNCANCC
  961 TGAATGGNGA ANGGNNNNCN NCNNGNANCN GCNCNNTNAN GCGNNGNNNG GNNNNNNNGN
1021 NNNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Yesbolatova, A., The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Nat. Commun. 11: 5701 (2020). PMID 33177522. [link to RRC of NBRP]