pNG_v2

价格:1000元

联系方式:I47-825O-882O

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载体基本信息

出品公司: 爱迪基因
载体名称: pNG_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: 32181
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

pNG_v2载体图谱



载体简介

	Transcription activator-like effectors (TALEs) are a class of naturally occurring DNA binding proteins that can be easily customized to recognize novel DNA sequences.The DNA binding domain of each TALE consists of tandem 34-amino acid repeat modules that can be rearranged according to a simple cipher to target new DNA sequences.
	This open source TALE toolbox provides a complete set of plasmids for building your own TALE nucleases (TALENs) and transcription factors (TALE-TFs) for genome editing and transcription modulation applications. The tandem repeats are PCR amplified from the monomer template plasmids and cloned into either a TALE-TF or TALEN backbone depending on the application. All of these plasmids are included in the TALE Toolbox kit and are depicted below:
	Customized TALENs and TALE-TFs can be constructed within one week using this kit, and the process can be easily scaled up for the construction of multiple TALEs in parallel.
	This toolbox and accompanying documentation allow one to efficiently assemble TALEN constructs with custom repeat arrays, containing up to 24 of these repeats. The reagents include a plasmid construct for making custom TAL effectors and one for TAL effector fusions to additional proteins of interest. More information and help can be found at TALeffector Resources Center .
	A 1-page rapid assembly protocol has been created by the depositor to be used with this kit. A detailed description of this toolbox as well as protocols for testing the activity of custom TALENs and TALE-TFs in mammalian cell lines is described in the following publication.

	A TAL Effector Toolbox for Genome Engineering
	Neville E. Sanjana, Le Cong, Yang Zhou, Margaret M. Cunniff, Guoping Feng and Feng Zhang
	Nature Protocols 7, 171-192 (2012). Open Access Article
	The Zhang lab has also deposited plasmids from the following paper:
	Efficient construction of sequence-specific TAL effectors for modulating mammalian transcription.
	Feng Zhang, Le Cong, Simona Lodato, Sriram Kosuri, George M. Church, and Paola Arlotta
	Nature Biotechnology, 29, 149-153 (2011). PUBMED. Open Access Article
	Please note that the plasmids from this paper and the TALE Toolbox are not equivalent. The new monomer NI, NG, NN, and HD that come with this kit are updated. This new set is compatible with the new primer design, which allow TALEs to be assembled in a more efficient manner (it also allows for the construction of longer TALEs). The new constructs also have bioinformatic support from the taleffectors.com website, so that users can assemble TALE sequence with the Zhang lab’s online resource.
	The most important similarity between the Nature Biotechnology constructs and the TALE Toolbox plasmids is that both versions were optimized for human expression. In this regard, they are similar in terms of expression efficiency, which is evident in the less than 0.01 difference between the two sets of TALE monomers when computing their codon preference for Homo sapiens.

载体序列

LOCUS       pNG_v2	2746 bp 	DNA	SYN
DEFINITION  pNG_v2
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..2746
                     /organism="pNG_v2"
                     /mol_type="other DNA"
     CDS             complement(0..809)
                     /label="ORF frame 3"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(12..809)
                     /label="KanR2 (variant)"
                     /gene="KanR2 (variant)"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(851..879)
                     /label="AmpR_promoter"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      990..1147
                     /label="rrnB_terminator"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(1748..1764)
                     /label="M13_forward20_primer"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(2019..2638)
                     /label="pBR322_origin"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2746..809)
                     /label="KanR2 (variant)"
                     /gene="KanR2 (variant)"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGK
                     PDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVD
                     ASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRV
                     GIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
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 CGGAGGGGGA 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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载体应用举例

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