pAC149-pCR8-dCas9VP160

价格:1200元

联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pAC149-pCR8-dCas9VP160 
出品公司: 爱迪基因
目录编号: 48221
存储实验室: Rudolf Jaenisch

载体骨架基本信息
载体名称: pCR8/GW/TOPO
载体出品公司: Invitrogen
载体抗性: 大观霉素
空载体大小: 2799 bp
完整质粒大小: 7442 bp
载体类型: CRISPR,网关的供体
高拷贝/低拷贝: 未知
生长条件: 37 ℃

插入基因信息
插入基因名称: dCas9(D10A;H840A) fusion with VP160 activation domain
插入大小: 4643 bp
基因突变: D10A;H840A nuclease-deficient
标签/融合蛋白: HA Tag(N端)            VP160(C端)
克隆方法: 限制性内切酶
5' 克隆位点: EcoRI(未破坏)
3' 克隆位点: EcoRI(未破坏)
5' 测序引物 M13F
3' 测序引物 M13R

文献引用方法
材料和方法部分: pAC149-pCR8-dCas9VP160 was a gift from Rudolf Jaenisch (Addgene plasmid # 48221)
参考文献部分: Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cheng AW, Wang H, Yang H, Shi L, Katz Y, Theunissen TW, Rangarajan S, Shivalila CS, Dadon DB, Jaenisch R. Cell Res. 2013 Aug 27. doi: 10.1038/cr.2013.122. 10.1038/cr.2013.122 PubMed 23979020

载体图谱质粒图谱和多克隆位点信息

pAC149-pCR8-dCas9VP160载体图谱

载体简介

载体序列

LOCUS       pAC149-pCR8-dCas9VP160	7442 bp 	DNA	SYN
DEFINITION  pAC149-pCR8-dCas9VP160
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..7442
                     /organism="pAC149-pCR8-dCas9VP160"
                     /mol_type="other DNA"
     terminator      complement(268..295)
                     /label="rrnB_T2_terminator"
     terminator      complement(427..470)
                     /label="rrnB_T1_terminator"
     misc_feature    522..544
                     /label="M13_pUC_fwd_primer"
     promoter        537..553
                     /label="M13_forward20_primer"
     misc_feature    complement(569..668)
                     /label="attL1"
     CDS             628..1080
                     /label="ORF frame 1"
     CDS             641..5305
                     /label="ORF frame 2"
     misc_feature    692..718
                     /label="HA"
     misc_feature    722..742
                     /label="SV40 NLS"
     misc_feature    728..742
                     /label="NLS"
     misc_feature    5321..5420
                     /label="attL2"
     misc_feature    5333..5420
                     /label="attL1"
     promoter        complement(5438..5456)
                     /label="T7_promoter"
     promoter        complement(5457..5475)
                     /label="M13_reverse_primer"
     gene            5689..6699
                     /label="spect"
                     /gene="spect"
     CDS             5689..6699
                     /label="ORF frame 1"
     rep_origin      6776..7395
                     /label="pBR322_origin"
ORIGIN
    1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA
   61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA
  121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA
  181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC
  241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA
  301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC
  361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA
  421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG
  481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA
  541 AACGACGGCC AGTCTTAAGC TCGGGCCCCA AATAATGATT TTATTTTGAC TGATAGTGAC
  601 CTGTTCGTTG CAACAAATTG ATGAGCAATG CTTTTTTATA ATGCCAACTT TGTACAAAAA
  661 AGCAGGCTCC GAATTcACCG GTgccaccAT GTACCCATAC GATGTTCCAG ATTACGCTTC
  721 GCCGAAGAAA AAGCGCAAGG TCGAAGCGTC CGACAAGAAG TACAGCATCG GCCTGGCCAT
  781 CGGCACCAAC TCTGTGGGCT GGGCCGTGAT CACCGACGAG TACAAGGTGC CCAGCAAGAA
  841 ATTCAAGGTG CTGGGCAACA CCGACCGGCA CAGCATCAAG AAGAACCTGA TCGGAGCCCT
  901 GCTGTTCGAC AGCGGCGAAA CAGCCGAGGC CACCCGGCTG AAGAGAACCG CCAGAAGAAG
  961 ATACACCAGA CGGAAGAACC GGATCTGCTA TCTGCAAGAG ATCTTCAGCA ACGAGATGGC
 1021 CAAGGTGGAC GACAGCTTCT TCCACAGACT GGAAGAGTCC TTCCTGGTGG AAGAGGATAA
 1081 GAAGCACGAG CGGCACCCCA TCTTCGGCAA CATCGTGGAC GAGGTGGCCT ACCACGAGAA
 1141 GTACCCCACC ATCTACCACC TGAGAAAGAA ACTGGTGGAC AGCACCGACA AGGCCGACCT
 1201 GCGGCTGATC TATCTGGCCC TGGCCCACAT GATCAAGTTC CGGGGCCACT TCCTGATCGA
 1261 GGGCGACCTG AACCCCGACA ACAGCGACGT GGACAAGCTG TTCATCCAGC TGGTGCAGAC
 1321 CTACAACCAG CTGTTCGAGG AAAACCCCAT CAACGCCAGC GGCGTGGACG CCAAGGCCAT
 1381 CCTGTCTGCC AGACTGAGCA AGAGCAGACG GCTGGAAAAT CTGATCGCCC AGCTGCCCGG
 1441 CGAGAAGAAG AATGGCCTGT TCGGCAACCT GATTGCCCTG AGCCTGGGCC TGACCCCCAA
 1501 CTTCAAGAGC AACTTCGACC TGGCCGAGGA TGCCAAACTG CAGCTGAGCA AGGACACCTA
 1561 CGACGACGAC CTGGACAACC TGCTGGCCCA GATCGGCGAC CAGTACGCCG ACCTGTTTCT
 1621 GGCCGCCAAG AACCTGTCCG ACGCCATCCT GCTGAGCGAC ATCCTGAGAG TGAACACCGA
 1681 GATCACCAAG GCCCCCCTGA GCGCCTCTAT GATCAAGAGA TACGACGAGC ACCACCAGGA
 1741 CCTGACCCTG CTGAAAGCTC TCGTGCGGCA GCAGCTGCCT GAGAAGTACA AAGAGATTTT
 1801 CTTCGACCAG AGCAAGAACG GCTACGCCGG CTACATTGAC GGCGGAGCCA GCCAGGAAGA
 1861 GTTCTACAAG TTCATCAAGC CCATCCTGGA AAAGATGGAC GGCACCGAGG AACTGCTCGT
 1921 GAAGCTGAAC AGAGAGGACC TGCTGCGGAA GCAGCGGACC TTCGACAACG GCAGCATCCC
 1981 CCACCAGATC CACCTGGGAG AGCTGCACGC CATTCTGCGG CGGCAGGAAG ATTTTTACCC
 2041 ATTCCTGAAG GACAACCGGG AAAAGATCGA GAAGATCCTG ACCTTCCGCA TCCCCTACTA
 2101 CGTGGGCCCT CTGGCCAGGG GAAACAGCAG ATTCGCCTGG ATGACCAGAA AGAGCGAGGA
 2161 AACCATCACC CCCTGGAACT TCGAGGAAGT GGTGGACAAG GGCGCTTCCG CCCAGAGCTT
 2221 CATCGAGCGG ATGACCAACT TCGATAAGAA CCTGCCCAAC GAGAAGGTGC TGCCCAAGCA
 2281 CAGCCTGCTG TACGAGTACT TCACCGTGTA TAACGAGCTG ACCAAAGTGA AATACGTGAC
 2341 CGAGGGAATG AGAAAGCCCG CCTTCCTGAG CGGCGAGCAG AAAAAGGCCA TCGTGGACCT
 2401 GCTGTTCAAG ACCAACCGGA AAGTGACCGT GAAGCAGCTG AAAGAGGACT ACTTCAAGAA
 2461 AATCGAGTGC TTCGACTCCG TGGAAATCTC CGGCGTGGAA GATCGGTTCA ACGCCTCCCT
 2521 GGGCACATAC CACGATCTGC TGAAAATTAT CAAGGACAAG GACTTCCTGG ACAATGAGGA
 2581 AAACGAGGAC ATTCTGGAAG ATATCGTGCT GACCCTGACA CTGTTTGAGG ACAGAGAGAT
 2641 GATCGAGGAA CGGCTGAAAA CCTATGCCCA CCTGTTCGAC GACAAAGTGA TGAAGCAGCT
 2701 GAAGCGGCGG AGATACACCG GCTGGGGCAG GCTGAGCCGG AAGCTGATCA ACGGCATCCG
 2761 GGACAAGCAG TCCGGCAAGA CAATCCTGGA TTTCCTGAAG TCCGACGGCT TCGCCAACAG
 2821 AAACTTCATG CAGCTGATCC ACGACGACAG CCTGACCTTT AAAGAGGACA TCCAGAAAGC
 2881 CCAGGTGTCC GGCCAGGGCG ATAGCCTGCA CGAGCACATT GCCAATCTGG CCGGCAGCCC
 2941 CGCCATTAAG AAGGGCATCC TGCAGACAGT GAAGGTGGTG GACGAGCTCG TGAAAGTGAT
 3001 GGGCCGGCAC AAGCCCGAGA ACATCGTGAT CGAAATGGCC AGAGAGAACC AGACCACCCA
 3061 GAAGGGACAG AAGAACAGCC GCGAGAGAAT GAAGCGGATC GAAGAGGGCA TCAAAGAGCT
 3121 GGGCAGCCAG ATCCTGAAAG AACACCCCGT GGAAAACACC CAGCTGCAGA ACGAGAAGCT
 3181 GTACCTGTAC TACCTGCAGA ATGGGCGGGA TATGTACGTG GACCAGGAAC TGGACATCAA
 3241 CCGGCTGTCC GACTACGATG TGGACGCCAT CGTGCCTCAG AGCTTTCTGA AGGACGACTC
 3301 CATCGACAAC AAGGTGCTGA CCAGAAGCGA CAAGAACCGG GGCAAGAGCG ACAACGTGCC
 3361 CTCCGAAGAG GTCGTGAAGA AGATGAAGAA CTACTGGCGG CAGCTGCTGA ACGCCAAGCT
 3421 GATTACCCAG AGAAAGTTCG ACAATCTGAC CAAGGCCGAG AGAGGCGGCC TGAGCGAACT
 3481 GGATAAGGCC GGCTTCATCA AGAGACAGCT GGTGGAAACC CGGCAGATCA CAAAGCACGT
 3541 GGCACAGATC CTGGACTCCC GGATGAACAC TAAGTACGAC GAGAATGACA AGCTGATCCG
 3601 GGAAGTGAAA GTGATCACCC TGAAGTCCAA GCTGGTGTCC GATTTCCGGA AGGATTTCCA
 3661 GTTTTACAAA GTGCGCGAGA TCAACAACTA CCACCACGCC CACGACGCCT ACCTGAACGC
 3721 CGTCGTGGGA ACCGCCCTGA TCAAAAAGTA CCCTAAGCTG GAAAGCGAGT TCGTGTACGG
 3781 CGACTACAAG GTGTACGACG TGCGGAAGAT GATCGCCAAG AGCGAGCAGG AAATCGGCAA
 3841 GGCTACCGCC AAGTACTTCT TCTACAGCAA CATCATGAAC TTTTTCAAGA CCGAGATTAC
 3901 CCTGGCCAAC GGCGAGATCC GGAAGCGGCC TCTGATCGAG ACAAACGGCG AAACCGGGGA
 3961 GATCGTGTGG GATAAGGGCC GGGATTTTGC CACCGTGCGG AAAGTGCTGA GCATGCCCCA
 4021 AGTGAATATC GTGAAAAAGA CCGAGGTGCA GACAGGCGGC TTCAGCAAAG AGTCTATCCT
 4081 GCCCAAGAGG AACAGCGATA AGCTGATCGC CAGAAAGAAG GACTGGGACC CTAAGAAGTA
 4141 CGGCGGCTTC GACAGCCCCA CCGTGGCCTA TTCTGTGCTG GTGGTGGCCA AAGTGGAAAA
 4201 GGGCAAGTCC AAGAAACTGA AGAGTGTGAA AGAGCTGCTG GGGATCACCA TCATGGAAAG
 4261 AAGCAGCTTC GAGAAGAATC CCATCGACTT TCTGGAAGCC AAGGGCTACA AAGAAGTGAA
 4321 AAAGGACCTG ATCATCAAGC TGCCTAAGTA CTCCCTGTTC GAGCTGGAAA ACGGCCGGAA
 4381 GAGAATGCTG GCCTCTGCCG GCGAACTGCA GAAGGGAAAC GAACTGGCCC TGCCCTCCAA
 4441 ATATGTGAAC TTCCTGTACC TGGCCAGCCA CTATGAGAAG CTGAAGGGCT CCCCCGAGGA
 4501 TAATGAGCAG AAACAGCTGT TTGTGGAACA GCACAAGCAC TACCTGGACG AGATCATCGA
 4561 GCAGATCAGC GAGTTCTCCA AGAGAGTGAT CCTGGCCGAC GCTAATCTGG ACAAAGTGCT
 4621 GTCCGCCTAC AACAAGCACC GGGATAAGCC CATCAGAGAG CAGGCCGAGA ATATCATCCA
 4681 CCTGTTTACC CTGACCAATC TGGGAGCCCC TGCCGCCTTC AAGTACTTTG ACACCACCAT
 4741 CGACCGGAAG AGGTACACCA GCACCAAAGA GGTGCTGGAC GCCACCCTGA TCCACCAGAG
 4801 CATCACCGGC CTGTACGAGA CACGGATCGA CCTGTCTCAG CTGGGAGGCG ACAGCCCCAA
 4861 GAAGAAGAGA AAGGTGGAGG CCAGCgggcc ggccGGATCC GGGCGCGCCG ACGCGCTGGA
 4921 CGATTTCGAT CTCGACATGC TGGGTTCTGA TGCCCTCGAT GACTTTGACC TGGATATGTT
 4981 GGGAAGCGAC GCATTGGATG ACTTTGATCT GGACATGCTC GGCTCCGATG CTCTGGACGA
 5041 TTTCGATCTC GATATGTTAg ggtcagacgc actggatgat ttcgaccttg atatgttgGG
 5101 AAGCGATGCC CTTGATGATT TCGACCTGGA CATGCTCggc agcgacgccc tggacgattt
 5161 cgatctggac atgctgGGGT CCGATGCCTT GGATGATTTT GACTTGGATA TGCTGgggag
 5221 tgatgccctg gacgactttg acctggacat gctgGGCTCC GATGCGCTCG ATGACTTCGA
 5281 TTTGGAtATG TTGTATATCG ATtgaTTAAT TAAGAATTCG ACCCAGCTTT CTTGTACAAA
 5341 GTTGGCATTA TAAAAAATAA TTGCTCATCA ATTTGTTGCA ACGAACAGGT CACTATCAGT
 5401 CAAAATAAAA TCATTATTTG CCATCCAGCT GATATCCCCT ATAGTGAGTC GTATTACATG
 5461 GTCATAGCTG TTTCCTGGCA GCTCTGGCCC GTGTCTCAAA ATCTCTGATG TTACATTGCA
 5521 CAAGATAAAA ATATATCATC ATGCCTCCTC TAGACCAGCC AGGACAGAAA TGCCTCGACT
 5581 TCGCTGCTGC CCAAGGTTGC CGGGTGACGC ACACCGTGGA AACGGATGAA GGCACGAACC
 5641 CAGTGGACAT AAGCCTGTTC GGTTCGTAAG CTGTAATGCA AGTAGCGTAT GCGCTCACGC
 5701 AACTGGTCCA GAACCTTGAC CGAACGCAGC GGTGGTAACG GCGCAGTGGC GGTTTTCATG
 5761 GCTTGTTATG ACTGTTTTTT TGGGGTACAG TCTATGCCTC GGGCATCCAA GCAGCAAGCG
 5821 CGTTACGCCG TGGGTCGATG TTTGATGTTA TGGAGCAGCA ACGATGTTAC GCAGCAGGGC
 5881 AGTCGCCCTA AAACAAAGTT AAACATCATG AGGGAAGCGG TGATCGCCGA AGTATCGACT
 5941 CAACTATCAG AGGTAGTTGG CGTCATCGAG CGCCATCTCG AACCGACGTT GCTGGCCGTA
 6001 CATTTGTACG GCTCCGCAGT GGATGGCGGC CTGAAGCCAC ACAGTGATAT TGATTTGCTG
 6061 GTTACGGTGA CCGTAAGGCT TGATGAAACA ACGCGGCGAG CTTTGATCAA CGACCTTTTG
 6121 GAAACTTCGG CTTCCCCTGG AGAGAGCGAG ATTCTCCGCG CTGTAGAAGT CACCATTGTT
 6181 GTGCACGACG ACATCATTCC GTGGCGTTAT CCAGCTAAGC GCGAACTGCA ATTTGGAGAA
 6241 TGGCAGCGCA ATGACATTCT TGCAGGTATC TTCGAGCCAG CCACGATCGA CATTGATCTG
 6301 GCTATCTTGC TGACAAAAGC AAGAGAACAT AGCGTTGCCT TGGTAGGTCC AGCGGCGGAG
 6361 GAACTCTTTG ATCCGGTTCC TGAACAGGAT CTATTTGAGG CGCTAAATGA AACCTTAACG
 6421 CTATGGAACT CGCCGCCCGA CTGGGCTGGC GATGAGCGAA ATGTAGTGCT TACGTTGTCC
 6481 CGCATTTGGT ACAGCGCAGT AACCGGCAAA ATCGCGCCGA AGGATGTCGC TGCCGACTGG
 6541 GCAATGGAGC GCCTGCCGGC CCAGTATCAG CCCGTCATAC TTGAAGCTAG ACAGGCTTAT
 6601 CTTGGACAAG AAGAAGATCG CTTGGCCTCG CGCGCAGATC AGTTGGAAGA ATTTGTCCAC
 6661 TACGTGAAAG GCGAGATCAC CAAGGTAGTC GGCAAATAAC CCTCGAGCCA CCCATGACCA
 6721 AAATCCCTTA ACGTGAGTTA CGCGTCGTTC CACTGAGCGT CAGACCCCGT AGAAAAGATC
 6781 AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT GCTGCTTGCA AACAAAAAAA
 6841 CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC TACCAACTCT TTTTCCGAAG
 6901 GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTCC TTCTAGTGTA GCCGTAGTTA
 6961 GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC TCGCTCTGCT AATCCTGTTA
 7021 CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG GGTTGGACTC AAGACGATAG
 7081 TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT CGTGCACACA GCCCAGCTTG
 7141 GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG AGCATTGAGA AAGCGCCACG
 7201 CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG GCAGGGTCGG AACAGGAGAG
 7261 CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT ATAGTCCTGT CGGGTTTCGC
 7321 CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG GGGGGCGGAG CCTATGGAAA
 7381 AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT GCTGGCCTTT TGCTCACATG
 7441 TT
//

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