FokI-dCas9

价格:1200元

联系方式:I47-825O-882O

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质粒基本信息

质粒名称: FokI-dCas9 
出品公司: 爱迪基因
目录编号: 52970
存储实验室: David Liu

载体骨架基本信息
载体名称: JDS246
空载体大小: 3400 bp
完整质粒大小: 8271 bp
载体类型: 哺乳细胞表达,CRISPR
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: Fok1 fused to dCas9
插入大小: 4800 bp
基因突变: D10A and H840A in Cas9
启动子 CMV
标签/融合蛋白: NLS(N端) Fok1核酸酶(同型二聚体野生型)(N端)
克隆方法: 结扎独立克隆
5' 测序引物 TAATACGACTCACTATAGG
3' 测序引物 CAGGGTCAAGGAAGGCACG

文献引用方法
材料和方法部分: FokI-dCas9 was a gift from David Liu (Addgene plasmid # 52970)
参考文献部分: Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Guilinger JP, Thompson DB, Liu DR. Nat Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2909. 10.1038/nbt.2909 PubMed 24770324

质粒图谱信息

FokI-dCas9载体图谱

质粒简介

质粒序列

LOCUS       FokI-dCas9	8271 bp 	DNA	SYN
DEFINITION  FokI-dCas9
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..8271
                     /organism="FokI-dCas9"
                     /mol_type="other DNA"
     misc_feature    286..306
                     /label="CMV_fwd_primer"
     promoter        287..356
                     /label="CMV_promoter"
     promoter        377..395
                     /label="T7_promoter"
     CDS             409..5280
                     /label="ORF frame 1"
     misc_feature    412..432
                     /label="FLAG"
     misc_feature    433..453
                     /label="FLAG"
     misc_feature    454..477
                     /label="FLAG"
     misc_feature    463..477
                     /label="EK"
     misc_feature    484..504
                     /label="SV40 NLS"
     misc_feature    484..507
                     /label="SV40 NLS"
     misc_feature    490..504
                     /label="NLS"
     misc_feature    complement(5326..5343)
                     /label="BGH_rev_primer"
     terminator      5329..5556
                     /label="bGH_PA_terminator"
     promoter        complement(5623..5641)
                     /label="M13_reverse_primer"
     misc_feature    complement(5640..5662)
                     /label="M13_pUC_rev_primer"
     promoter        complement(5676..5705)
                     /label="lac_promoter"
     rep_origin      complement(6014..6633)
                     /label="pBR322_origin"
     gene            complement(6788..7648)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(6788..7648)
                     /label="ORF frame 3"
     promoter        complement(7690..7718)
                     /label="AmpR_promoter"
     promoter        8024..329
                     /label="CMV_immearly_promoter"
     misc_feature    8103..119
                     /label="CAG_enhancer"
ORIGIN
    1 ATATGCCAAG TACGCCCCCT ATTGACGTCA ATGACGGTAA ATGGCCCGCC TGGCATTATG
   61 CCCAGTACAT GACCTTATGG GACTTTCCTA CTTGGCAGTA CATCTACGTA TTAGTCATCG
  121 CTATTACCAT GGTGATGCGG TTTTGGCAGT ACATCAATGG GCGTGGATAG CGGTTTGACT
  181 CACGGGGATT TCCAAGTCTC CACCCCATTG ACGTCAATGG GAGTTTGTTT TGGCACCAAA
  241 ATCAACGGGA CTTTCCAAAA TGTCGTAACA ACTCCGCCCC ATTGACGCAA ATGGGCGGTA
  301 GGCGTGTACG GTGGGAGGTC TATATAAGCA GAGCTGGTTT AGTGAACCGT CAGATCCGCT
  361 AGAGATCCGC GGCCGCTAAT ACGACTCACT ATAGGGAGAG CCGCCACCAT GGACTACAAA
  421 GACCATGACG GTGATTATAA AGATCATGAC ATCGATTACA AGGATGACGA TGACAAGATG
  481 GCCCCCAAGA AGAAGAGGAA GGTGGGCATT CACCGCGGGG TACCTGGAGG TTCTGGATCC
  541 CAACTAGTCA AAAGTGAACT GGAGGAGAAG AAATCTGAAC TTCGTCATAA ATTGAAATAT
  601 GTGCCTCATG AATATATTGA ATTAATTGAA ATTGCCAGAA ATTCCACTCA GGATAGAATT
  661 CTTGAAATGA AGGTAATGGA ATTTTTTATG AAAGTTTATG GATATAGAGG TAAACATTTG
  721 GGTGGATCAA GGAAACCGGA CGGAGCAATT TATACTGTCG GATCTCCTAT TGATTACGGT
  781 GTGATCGTGG ATACTAAAGC TTATAGCGGA GGTTATAATC TGCCAATTGG CCAAGCAGAT
  841 GAAATGCAAC GATATGTCGA AGAAAATCAA ACACGAAACA AACATATCAA CCCTAATGAA
  901 TGGTGGAAAG TCTATCCATC TTCTGTAACG GAATTTAAGT TTTTATTTGT GAGTGGTCAC
  961 TTTAAAGGAA ACTACAAAGC TCAGCTTACA CGATTAAATC ATATCACTAA TTGTAATGGA
 1021 GCTGTTCTTA GTGTAGAAGA GCTTTTAATT GGTGGAGAAA TGATTAAAGC CGGCACATTA
 1081 ACCTTAGAGG AAGTCAGACG GAAATTTAAT AACGGCGAGA TAAACTTTAG CGGCAGCGAG
 1141 ACTCCCGGGA CCTCAGAGTC CGCCACACCC GAAAGTGATA AAAAGTATTC TATTGGTTTA
 1201 GCTATCGGCA CTAATTCCGT TGGATGGGCT GTCATAACCG ATGAATACAA AGTACCTTCA
 1261 AAGAAATTTA AGGTGTTGGG GAACACAGAC CGTCATTCGA TTAAAAAGAA TCTTATCGGT
 1321 GCCCTCCTAT TCGATAGTGG CGAAACGGCA GAGGCGACTC GCCTGAAACG AACCGCTCGG
 1381 AGAAGGTATA CACGTCGCAA GAACCGAATA TGTTACTTAC AAGAAATTTT TAGCAATGAG
 1441 ATGGCCAAAG TTGACGATTC TTTCTTTCAC CGTTTGGAAG AGTCCTTCCT TGTCGAAGAG
 1501 GACAAGAAAC ATGAACGGCA CCCCATCTTT GGAAACATAG TAGATGAGGT GGCATATCAT
 1561 GAAAAGTACC CAACGATTTA TCACCTCAGA AAAAAGCTAG TTGACTCAAC TGATAAAGCG
 1621 GACCTGAGGT TAATCTACTT GGCTCTTGCC CATATGATAA AGTTCCGTGG GCACTTTCTC
 1681 ATTGAGGGTG ATCTAAATCC GGACAACTCG GATGTCGACA AACTGTTCAT CCAGTTAGTA
 1741 CAAACCTATA ATCAGTTGTT TGAAGAGAAC CCTATAAATG CAAGTGGCGT GGATGCGAAG
 1801 GCTATTCTTA GCGCCCGCCT CTCTAAATCC CGACGGCTAG AAAACCTGAT CGCACAATTA
 1861 CCCGGAGAGA AGAAAAATGG GTTGTTCGGT AACCTTATAG CGCTCTCACT AGGCCTGACA
 1921 CCAAATTTTA AGTCGAACTT CGACTTAGCT GAAGATGCCA AATTGCAGCT TAGTAAGGAC
 1981 ACGTACGATG ACGATCTCGA CAATCTACTG GCACAAATTG GAGATCAGTA TGCGGACTTA
 2041 TTTTTGGCTG CCAAAAACCT TAGCGATGCA ATCCTCCTAT CTGACATACT GAGAGTTAAT
 2101 ACTGAGATTA CCAAGGCGCC GTTATCCGCT TCAATGATCA AAAGGTACGA TGAACATCAC
 2161 CAAGACTTGA CACTTCTCAA GGCCCTAGTC CGTCAGCAAC TGCCTGAGAA ATATAAGGAA
 2221 ATATTCTTTG ATCAGTCGAA AAACGGGTAC GCAGGTTATA TTGACGGCGG AGCGAGTCAA
 2281 GAGGAATTCT ACAAGTTTAT CAAACCCATA TTAGAGAAGA TGGATGGGAC GGAAGAGTTG
 2341 CTTGTAAAAC TCAATCGCGA AGATCTACTG CGAAAGCAGC GGACTTTCGA CAACGGTAGC
 2401 ATTCCACATC AAATCCACTT AGGCGAATTG CATGCTATAC TTAGAAGGCA GGAGGATTTT
 2461 TATCCGTTCC TCAAAGACAA TCGTGAAAAG ATTGAGAAAA TCCTAACCTT TCGCATACCT
 2521 TACTATGTGG GACCCCTGGC CCGAGGGAAC TCTCGGTTCG CATGGATGAC AAGAAAGTCC
 2581 GAAGAAACGA TTACTCCATG GAATTTTGAG GAAGTTGTCG ATAAAGGTGC GTCAGCTCAA
 2641 TCGTTCATCG AGAGGATGAC CAACTTTGAC AAGAATTTAC CGAACGAAAA AGTATTGCCT
 2701 AAGCACAGTT TACTTTACGA GTATTTCACA GTGTACAATG AACTCACGAA AGTTAAGTAT
 2761 GTCACTGAGG GCATGCGTAA ACCCGCCTTT CTAAGCGGAG AACAGAAGAA AGCAATAGTA
 2821 GATCTGTTAT TCAAGACCAA CCGCAAAGTG ACAGTTAAGC AATTGAAAGA GGACTACTTT
 2881 AAGAAAATTG AATGCTTCGA TTCTGTCGAG ATCTCCGGGG TAGAAGATCG ATTTAATGCG
 2941 TCACTTGGTA CGTATCATGA CCTCCTAAAG ATAATTAAAG ATAAGGACTT CCTGGATAAC
 3001 GAAGAGAATG AAGATATCTT AGAAGATATA GTGTTGACTC TTACCCTCTT TGAAGATCGG
 3061 GAAATGATTG AGGAAAGACT AAAAACATAC GCTCACCTGT TCGACGATAA GGTTATGAAA
 3121 CAGTTAAAGA GGCGTCGCTA TACGGGCTGG GGACGATTGT CGCGGAAACT TATCAACGGG
 3181 ATAAGAGACA AGCAAAGTGG TAAAACTATT CTCGATTTTC TAAAGAGCGA CGGCTTCGCC
 3241 AATAGGAACT TTATGCAGCT GATCCATGAT GACTCTTTAA CCTTCAAAGA GGATATACAA
 3301 AAGGCACAGG TTTCCGGACA AGGGGACTCA TTGCACGAAC ATATTGCGAA TCTTGCTGGT
 3361 TCGCCAGCCA TCAAAAAGGG CATACTCCAG ACAGTCAAAG TAGTGGATGA GCTAGTTAAG
 3421 GTCATGGGAC GTCACAAACC GGAAAACATT GTAATCGAGA TGGCACGCGA AAATCAAACG
 3481 ACTCAGAAGG GGCAAAAAAA CAGTCGAGAG CGGATGAAGA GAATAGAAGA GGGTATTAAA
 3541 GAACTGGGCA GCCAGATCTT AAAGGAGCAT CCTGTGGAAA ATACCCAATT GCAGAACGAG
 3601 AAACTTTACC TCTATTACCT ACAAAATGGA AGGGACATGT ATGTTGATCA GGAACTGGAC
 3661 ATAAACCGTT TATCTGATTA CGACGTCGAT GCCATTGTAC CCCAATCCTT TTTGAAGGAC
 3721 GATTCAATCG ACAATAAAGT GCTTACACGC TCGGATAAGA ACCGAGGGAA AAGTGACAAT
 3781 GTTCCAAGCG AGGAAGTCGT AAAGAAAATG AAGAACTATT GGCGGCAGCT CCTAAATGCG
 3841 AAACTGATAA CGCAAAGAAA GTTCGATAAC TTAACTAAAG CTGAGAGGGG TGGCTTGTCT
 3901 GAACTTGACA AGGCCGGATT TATTAAACGT CAGCTCGTGG AAACCCGCCA AATCACAAAG
 3961 CATGTTGCAC AGATACTAGA TTCCCGAATG AATACGAAAT ACGACGAGAA CGATAAGCTG
 4021 ATTCGGGAAG TCAAAGTAAT CACTTTAAAG TCAAAATTGG TGTCGGACTT CAGAAAGGAT
 4081 TTTCAATTCT ATAAAGTTAG GGAGATAAAT AACTACCACC ATGCGCACGA CGCTTATCTT
 4141 AATGCCGTCG TAGGGACCGC ACTCATTAAG AAATACCCGA AGCTAGAAAG TGAGTTTGTG
 4201 TATGGTGATT ACAAAGTTTA TGACGTCCGT AAGATGATCG CGAAAAGCGA ACAGGAGATA
 4261 GGCAAGGCTA CAGCCAAATA CTTCTTTTAT TCTAACATTA TGAATTTCTT TAAGACGGAA
 4321 ATCACTCTGG CAAACGGAGA GATACGCAAA CGACCTTTAA TTGAAACCAA TGGGGAGACA
 4381 GGTGAAATCG TATGGGATAA GGGCCGGGAC TTCGCGACGG TGAGAAAAGT TTTGTCCATG
 4441 CCCCAAGTCA ACATAGTAAA GAAAACTGAG GTGCAGACCG GAGGGTTTTC AAAGGAATCG
 4501 ATTCTTCCAA AAAGGAATAG TGATAAGCTC ATCGCTCGTA AAAAGGACTG GGACCCGAAA
 4561 AAGTACGGTG GCTTCGATAG CCCTACAGTT GCCTATTCTG TCCTAGTAGT GGCAAAAGTT
 4621 GAGAAGGGAA AATCCAAGAA ACTGAAGTCA GTCAAAGAAT TATTGGGGAT AACGATTATG
 4681 GAGCGCTCGT CTTTTGAAAA GAACCCCATC GACTTCCTTG AGGCGAAAGG TTACAAGGAA
 4741 GTAAAAAAGG ATCTCATAAT TAAACTACCA AAGTATAGTC TGTTTGAGTT AGAAAATGGC
 4801 CGAAAACGGA TGTTGGCTAG CGCCGGAGAG CTTCAAAAGG GGAACGAACT CGCACTACCG
 4861 TCTAAATACG TGAATTTCCT GTATTTAGCG TCCCATTACG AGAAGTTGAA AGGTTCACCT
 4921 GAAGATAACG AACAGAAGCA ACTTTTTGTT GAGCAGCACA AACATTATCT CGACGAAATC
 4981 ATAGAGCAAA TTTCGGAATT CAGTAAGAGA GTCATCCTAG CTGATGCCAA TCTGGACAAA
 5041 GTATTAAGCG CATACAACAA GCACAGGGAT AAACCCATAC GTGAGCAGGC GGAAAATATT
 5101 ATCCATTTGT TTACTCTTAC CAACCTCGGC GCTCCAGCCG CATTCAAGTA TTTTGACACA
 5161 ACGATAGATC GCAAACGATA CACTTCTACC AAGGAGGTGC TAGACGCGAC ACTGATTCAC
 5221 CAATCCATCA CGGGATTATA TGAAACTCGG ATAGATTTGT CACAGCTTGG GGGTGACTGA
 5281 CCGGTCATCA TCACCATCAC CATTGAGTTT AAACCCGCTG ATCAGCCTCG ACTGTGCCTT
 5341 CTAGTTGCCA GCCATCTGTT GTTTGCCCCT CCCCCGTGCC TTCCTTGACC CTGGAAGGTG
 5401 CCACTCCCAC TGTCCTTTCC TAATAAAATG AGGAAATTGC ATCGCATTGT CTGAGTAGGT
 5461 GTCATTCTAT TCTGGGGGGT GGGGTGGGGC AGGACAGCAA GGGGGAGGAT TGGGAAGACA
 5521 ATAGCAGGCA TGCTGGGGAT GCGGTGGGCT CTATGGCTTC TGAGGCGGAA AGAACCAGCT
 5581 GGGGCTCGAT ACCGTCGACC TCTAGCTAGA GCTTGGCGTA ATCATGGTCA TAGCTGTTTC
 5641 CTGTGTGAAA TTGTTATCCG CTCACAATTC CACACAACAT ACGAGCCGGA AGCATAAAGT
 5701 GTAAAGCCTA GGGTGCCTAA TGAGTGAGCT AACTCACATT AATTGCGTTG CGCTCACTGC
 5761 CCGCTTTCCA GTCGGGAAAC CTGTCGTGCC AGCTGCATTA ATGAATCGGC CAACGCGCGG
 5821 GGAGAGGCGG TTTGCGTATT GGGCGCTCTT CCGCTTCCTC GCTCACTGAC TCGCTGCGCT
 5881 CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAA GGCGGTAATA CGGTTATCCA
 5941 CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA
 6001 ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCT CCGCCCCCCT GACGAGCATC
 6061 ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGAC AGGACTATAA AGATACCAGG
 6121 CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCC GACCCTGCCG CTTACCGGAT
 6181 ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTC TCAATGCTCA CGCTGTAGGT
 6241 ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTG TGTGCACGAA CCCCCCGTTC
 6301 AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGA GTCCAACCCG GTAAGACACG
 6361 ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAG CAGAGCGAGG TATGTAGGCG
 6421 GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTA CACTAGAAGG ACAGTATTTG
 6481 GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAG AGTTGGTAGC TCTTGATCCG
 6541 GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTG CAAGCAGCAG ATTACGCGCA
 6601 GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA
 6661 ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATC AAAAAGGATC TTCACCTAGA
 6721 TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAG TATATATGAG TAAACTTGGT
 6781 CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTC AGCGATCTGT CTATTTCGTT
 6841 CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTAC GATACGGGAG GGCTTACCAT
 6901 CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTC ACCGGCTCCA GATTTATCAG
 6961 CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGG TCCTGCAACT TTATCCGCCT
 7021 CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAG TAGTTCGCCA GTTAATAGTT
 7081 TGCGCAACGT TGTTGCCATT GCTACAGGCA TCGTGGTGTC ACGCTCGTCG TTTGGTATGG
 7141 CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTAC ATGATCCCCC ATGTTGTGCA
 7201 AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT
 7261 TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTAC TGTCATGCCA TCCGTAAGAT
 7321 GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTG AGAATAGTGT ATGCGGCGAC
 7381 CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGC GCCACATAGC AGAACTTTAA
 7441 AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACT CTCAAGGATC TTACCGCTGT
 7501 TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTG ATCTTCAGCA TCTTTTACTT
 7561 TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAA TGCCGCAAAA AAGGGAATAA
 7621 GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTT TCAATATTAT TGAAGCATTT
 7681 ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA
 7741 TAGGGGTTCC GCGCACATTT CCCCGAAAAG TGCCACCTGA CGTCGACGGA TCGGGAGATC
 7801 GATCTCCCGA TCCCCTAGGG TCGACTCTCA GTACAATCTG CTCTGATGCC GCATAGTTAA
 7861 GCCAGTATCT GCTCCCTGCT TGTGTGTTGG AGGTCGCTGA GTAGTGCGCG AGCAAAATTT
 7921 AAGCTACAAC AAGGCAAGGC TTGACCGACA ATTGCATGAA GAATCTGCTT AGGGTTAGGC
 7981 GTTTTGCGCT GCTTCGCGAT GTACGGGCCA GATATACGCG TTGACATTGA TTATTGACTA
 8041 GTTATTAATA GTAATCAATT ACGGGGTCAT TAGTTCATAG CCCATATATG GAGTTCCGCG
 8101 TTACATAACT TACGGTAAAT GGCCCGCCTG GCTGACCGCC CAACGACCCC CGCCCATTGA
 8161 CGTCAATAAT GACGTATGTT CCCATAGTAA CGCCAATAGG GACTTTCCAT TGACGTCAAT
 8221 GGGTGGACTA TTTACGGTAA ACTGCCCACT TGGCAGTACA TCAAGTGTAT C

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