pNN_v2载体

pNN_v2载体

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  • ¥1000
  • ZYbscience
  • 中国/美国
  • ZY32182
  • 2025年07月16日
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    • 详细信息
    • 技术资料
    • 保存条件

      -20℃低温保存

    • 保质期

      三年

    • 英文名

      pNN_v2

    • 库存

      20

    • 供应商

      泽叶生物

    载体基本信息

    出品公司: ZYbscience
    载体名称: pNN_v2
    质粒类型: 靶向基因操作载体
    高拷贝/低拷贝: 高拷贝
    启动子: --
    克隆方法: 多克隆位点,限制性内切酶
    载体大小: 2746bp
    5' 测序引物及序列: --
    3' 测序引物及序列: --
    载体标签: --
    载体抗性: 卡纳霉素(Kanamycin)
    筛选标记: --
    备注: 克隆菌株为Stbl3。
    产品目录号: 32182
    稳定性: --
    组成型: --
    病毒/非病毒: --

    载体质粒图谱和多克隆位点信息

    pNN_v2载体图谱


    载体简介

    (a) Natural structure of TALEs derived from Xanthomonas sp. Each DNA-binding module consists of 34 amino acids, where the 
    RVDs in the 12th and 13th amino acid positions of each repeat specify the DNA base being targeted according to the cipher NG = T, HD = C, NI = A, and NN = G or A. The DNA-binding modules are flanked by nonrepetitive N and C termini, which carry the 
    translocation, nuclear localization (NLS) and transcription activation (AD) domains. A cryptic signal within the N terminus 
    specifies a thymine as the first base of the target site.
    (b) The TALE toolbox allows rapid and inexpensive construction of custom TALE-TFs and TALENs. The kit consists of 12 plasmids 
    in total: four monomer plasmids to be used as templates for PCR amplification, four TALE-TF and four TALEN cloning backbones 
    corresponding to four different bases targeted by the 0.5 repeat. CMV, cytomegalovirus promoter; N term, nonrepetitive N 
    terminus from the Hax3 TALE; C term, nonrepetitive C terminus from the Hax3 TALE; BsaI, type IIs restriction sites used for 
    the insertion of custom TALE DNA-binding domains; ccdB + CmR, negative selection cassette containing the ccdB negative 
    selection gene and chloramphenicol resistance gene; NLS, nuclear localization signal; VP64, synthetic transcriptional 
    activator derived from VP16 protein of herpes simplex virus; 2A, 2A self-cleavage linker; EGFP, enhanced green fluorescent 
    protein; polyA signal, polyadenylation signal; FokI, catalytic domain from the FokI endonuclease. 
    (c) TALEs can be used to generate custom TALE-TFs and modulate the transcription of endogenous genes from the genome. The 
    TALE DNA-binding domain is fused to the synthetic VP64 transcriptional activator, which recruits RNA polymerase and other factors needed to initiate transcription. 
    (d) TALENs can be used to generate site-specific double-strand breaks to facilitate genome editing through nonhomologous 
    repair or homology directed repair. Two TALENs target a pair of binding sites flanking a 16-bp spacer. The left and right 
    TALENs recognize the top and bottom strands of the target sites, respectively. Each TALE DNA-binding domain is fused to the 
    catalytic domain of FokI endonuclease; when FokI dimerizes, it cuts the DNA in the region between the left and right 
    TALEN-binding sites.
    

    载体序列

    LOCUS       pNN_v2	2746 bp 	DNA	SYN
    DEFINITION  pNN_v2
    ACCESSION   
    KEYWORDS    
    SOURCE      
      ORGANISM  other sequences; artificial sequences; vectors.
    FEATURES             Location/Qualifiers
         source          1..2746
                         /organism="pNN_v2"
                         /mol_type="other DNA"
         CDS             complement(0..809)
                         /label="ORF frame 3"
                       
         gene            complement(12..809)
                         /label="KanR2 (variant)"
                         /gene="KanR2 (variant)"
                       
         promoter        complement(851..879)
                         /label="AmpR_promoter"
                         
         terminator      990..1147
                         /label="rrnB_terminator"
                        
         promoter        complement(1748..1764)
                         /label="M13_forward20_primer"
                        
         rep_origin      complement(2019..2638)
                         /label="pBR322_origin"
               
         gene            complement(2746..809)
                         /label="KanR2 (variant)"
                         /gene="KanR2 (variant)"
         
    ORIGIN
        1 TAGAAAAACT CATCGAGCAT CAAATGAAAC TGCAATTTAT TCATATCAGG ATTATCAATA
       61 CCATATTTTT GAAAAAGCCG TTTCTGTAAT GAAGGAGAAA ACTCACCGAG GCAGTTCCAT
      121 AGGATGGCAA GATCCTGGTA TCGGTCTGCG ATTCCGACTC GTCCAACATC AATACAACCT
      181 ATTAATTTCC CCTCGTCAAA AATAAGGTTA TCAAGTGAGA AATCACCATG AGTGACGACT
      241 GAATCCGGTG AGAATGGCAA AAGTTTATGC ATTTCTTTCC AGACTTGTTC AACAGGCCAG
      301 CCATTACGCT CGTCATCAAA ATCACTCGCA TCAACCAAAC CGTTATTCAT TCGTGATTGC
      361 GCCTGAGCGA GGCGAAATAC GCGATCGCTG TTAAAAGGAC AATTACAAAC AGGAATCGAG
      421 TGCAACCGGC GCAGGAACAC TGCCAGCGCA TCAACAATAT TTTCACCTGA ATCAGGATAT
      481 TCTTCTAATA CCTGGAACGC TGTTTTTCCG GGGATCGCAG TGGTGAGTAA CCATGCATCA
      541 TCAGGAGTAC GGATAAAATG CTTGATGGTC GGAAGTGGCA TAAATTCCGT CAGCCAGTTT
      601 AGTCTGACCA TCTCATCTGT AACATCATTG GCAACGCTAC CTTTGCCATG TTTCAGAAAC
      661 AACTCTGGCG CATCGGGCTT CCCATACAAG CGATAGATTG TCGCACCTGA TTGCCCGACA
      721 TTATCGCGAG CCCATTTATA CCCATATAAA TCAGCATCCA TGTTGGAATT TAATCGCGGC
      781 CTCGACGTTT CCCGTTGAAT ATGGCTCATA TTCTTCCTTT TTCAATATTA TTGAAGCATT
      841 TATCAGGGTT ATTGTCTCAT GAGCGGATAC ATATTTGAAT GTATTTAGAA AAATAAACAA
      901 ATAGGGGTCA GTGTTACAAC CAATTAACCA ATTCTGAACA TTATCGCGAG CCCATTTATA
      961 CCTGAATATG GCTCATAACA CCCCTTGTTT GCCTGGCGGC AGTAGCGCGG TGGTCCCACC
     1021 TGACCCCATG CCGAACTCAG AAGTGAAACG CCGTAGCGCC GATGGTAGTG TGGGGACTCC
     1081 CCATGCGAGA GTAGGGAACT GCCAGGCATC AAATAAAACG AAAGGCTCAG TCGAAAGACT
     1141 GGGCCTTTCG CCCGGGCTAA TTAGGGGGTG TCGCCCTTCG CTGAACTCAC CCCAGAGCAG
     1201 GTCGTGGCAA TTGCGAGCAA CAACGGGGGA AAGCAGGCAC TCGAAACCGT CCAGAGGTTG
     1261 CTGCCTGTGC TGTGCCAAGC GCACGGACGT CAAAAGGGCG ACACAAAATT TATTCTAAAT
     1321 GCATAATAAA TACTGATAAC ATCTTATAGT TTGTATTATA TTTTGTATTA TCGTTGACAT
     1381 GTATAATTTT GATATCAAAA ACTGATTTTC CCTTTATTAT TTTCGAGATT TATTTTCTTA
     1441 ATTCTCTTTA ACAAACTAGA AATATTGTAT ATACAAAAAA TCATAAATAA TAGATGAATA
     1501 GTTTAATTAT AGGTGTTCAT CAATCGAAAA AGCAACGTAT CTTATTTAAA GTGCGTTGCT
     1561 TTTTTCTCAT TTATAAGGTT AAATAATTCT CATATATCAA GCAAAGTGAC AGGCGCCCTT
     1621 AAATATTCTG ACAAATGCTC TTTCCCTAAA CTCCCCCCAT AAAAAAACCC GCCGAAGCGG
     1681 GTTTTTACGT TATTTGCGGA TTAACGATTA CTCGTTATCA GAACCGCCCA GGGGGCCCGA
     1741 GCTTAAGACT GGCCGTCGTT TTACAACACA GAAAGAGTTT GTAGAAACGC AAAAAGGCCA
     1801 TCCGTCAGGG GCCTTCTGCT TAGTTTGATG CCTGGCAGTT CCCTACTCTC GCCTTCCGCT
     1861 TCCTCGCTCA CTGACTCGCT GCGCTCGGTC GTTCGGCTGC GGCGAGCGGT ATCAGCTCAC
     1921 TCAAAGGCGG TAATACGGTT ATCCACAGAA TCAGGGGATA ACGCAGGAAA GAACATGTGA
     1981 GCAAAAGGCC AGCAAAAGGC CAGGAACCGT AAAAAGGCCG CGTTGCTGGC GTTTTTCCAT
     2041 AGGCTCCGCC CCCCTGACGA GCATCACAAA AATCGACGCT CAAGTCAGAG GTGGCGAAAC
     2101 CCGACAGGAC TATAAAGATA CCAGGCGTTT CCCCCTGGAA GCTCCCTCGT GCGCTCTCCT
     2161 GTTCCGACCC TGCCGCTTAC CGGATACCTG TCCGCCTTTC TCCCTTCGGG AAGCGTGGCG
     2221 CTTTCTCATA GCTCACGCTG TAGGTATCTC AGTTCGGTGT AGGTCGTTCG CTCCAAGCTG
     2281 GGCTGTGTGC ACGAACCCCC CGTTCAGCCC GACCGCTGCG CCTTATCCGG TAACTATCGT
     2341 CTTGAGTCCA ACCCGGTAAG ACACGACTTA TCGCCACTGG CAGCAGCCAC TGGTAACAGG
     2401 ATTAGCAGAG CGAGGTATGT AGGCGGTGCT ACAGAGTTCT TGAAGTGGTG GGCTAACTAC
     2461 GGCTACACTA GAAGAACAGT ATTTGGTATC TGCGCTCTGC TGAAGCCAGT TACCTTCGGA
     2521 AAAAGAGTTG GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT
     2581 GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC TTTGATCTTT
     2641 TCTACGGGGT CTGACGCTCA GTGGAACGAC GCGCGCGTAA CTCACGTTAA GGGATTTTGG
     2701 TCATGAGCTT GCGCCGTCCC GTCAAGTCAG CGTAATGCTC TGCTTT
    //

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