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  • pGuide-IT-tdTomato

    價格:8000元

    聯系方式:I47-825O-882O

    買家導航

    載體基本信息

    出品公司: Clontech
    載體名稱: pGuide-it-tdTomato
    質粒類型: 慢病毒載體;CRISPR/Cas9載體;熒光報告載體
    高拷貝/低拷貝: 高拷貝
    克隆方法: 限制性內切酶,多克隆位點
    啟動子: Minimal CMV
    載體大小: 9038 bp
    5' 測序引物及序列: --
    3' 測序引物及序列: --
    載體標簽: --
    載體抗性: 氨芐青霉素
    篩選標記: tdTomato紅色熒光蛋白
    克隆菌株: Stbl3、Stellar
    宿主細胞(系): 常規細胞系(293、CV-1、CHO等)
    備注: 慢病毒載體pGuide-it-tdTomato是CRISPR/Cas9載體,Clontech提供的是線性化的質粒,Biofeng提供的是環狀質粒。
    產品目錄號: 632604
    穩定性: 穩表達
    組成型/誘導型: 組成型
    病毒/非病毒: 慢病毒

    載體質粒圖譜和多克隆位點信息

    pGuide-it-tdTomato載體圖譜



    pGuide-it-tdTomato 多克隆位點

    pGuide-it-tdTomato 載體特征

    載體簡介

    
    
    CRISPR/Cas9 技術
    
    The Guide-it CRISPR/Cas9 System (Green, Cat. No. 632601; Red, Cat. No. 632602) is a complete system for the cloning and expression of a target single guide RNA (sgRNA) for mammalian gene modification studies using CRISPR/Cas9 technology. The included vector is used to simultaneously express Cas9, a target-specific sgRNA, and an exceptionally bright fluorescent protein (either ZsGreen1 or tdTomato) for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry. The vector is pre-linearized for simple insertion of a target sequence for sgRNA expression from the human U6 promoter; Cas9 nuclease and either ZsGreen1 or tdTomato are co-expressed from a bidirectional CMV promoter. Sufficient reagents are provided in this kit for construction of 10 different target (sgRNA) expression plasmids. To construct the vector, a pair of user-provided oligos corresponding to the target genomic sequence of interest are annealed to form a duplex. Then, the duplexed DNA is cloned into the pre-linearized vector using the included high-efficiency ligation mix. This kit also contains necessary controls and Stellar? Competent Cells.
    
    Cloning and Expression System for sgRNAs
    The Guide-it CRISPR/Cas9 Systems are kits for the cloning and expression of target single guide RNAs (sgRNAs) for mammalian genome editing using CRISPR/Cas9 technology. The vector in this system simultaneously expresses Cas9 nuclease, a target-specific sgRNA, and an exceptionally bright fluorescent protein for monitoring transfection efficiency and/or for further enriching/isolating transfected cells by flow cytometry (ZsGreen1 and tdTomato versions are available). Generating a plasmid that expresses a sequence-specific sgRNA with this system is simple; a pair of user-provided oligos corresponding to the target genomic sequence of interest are annealed to form a duplex, and the duplexed oligos are inserted into the pre-linearized vector using the included high-efficiency ligation mix. The kit also includes Stellar competent cells to ensure high efficiency transformation.
    
    Features
    A single vector expresses your target-specific sgRNA from the human U6 promoter, and Cas9 nuclease and a bright fluorescent marker (either ZsGreen1 or tdTomato) are expressed from a bidirectional CMV promoter tdTomato and ZsGreen1 are exceptionally bright fluorescent proteins, 6x and 2.5x brighter than EGFP, respectively; these markers can be used to monitor transfection  efficiency and/or to enrich/isolate transfected cells by flow cytometry Simple plasmid construction—design oligos specific to your target sequence, and clone into the pre-linearized plasmid using the included ligation reagents and high- efficiency competent cells Includes sufficient reagents for construction of 10 different target (sgRNA) expression plasmids
    
    Applications
    Cloning and expression of a target single guide RNA (sgRNA) for mammalian genome editing using CRISPR/Cas9 technology
    
    

    載體序列

    LOCUS       pGuide-it-tdTomato Vector (Linear)           9042 bp ds-DNA    linear   SYN 05-DEC-2014
    COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
    COMMENT     Cat No. 632602, 632604
    SOURCE      synthetic DNA construct
    FEATURES             Location/Qualifiers
         misc_feature    1..93
                         /label="sgRNA"
                         /note="single-guide RNA scaffold segment; user clones guide sequence"
         rep_origin    complement(147..746)
                         /label="pUC origin"
                         /note="ori;high-copy-number ColE1/pMB1/pBR322/pUC origin of
                         replication"
         CDS             complement(908..1768)
                         /label="AmpR"
                         /note="beta-lactamase, confers resistance to ampicillin, carbenicillin, and
                         related antibiotics"
         polyA_signal    complement(1904..2091)
                         /label="SV40 polyA signal"
         CDS             complement(2129..3556)
                         /label="tdTomato"
         promoter        complement(3574..3642)
                         /label="Minimal CMV Promoter 2"
         misc_feature    3706..4115
                         /label="Enhancer"
         promoter        4116..4241
                         /label="Minimal CMV Promoter 1"
         CDS             4286..8389
                         /label="Cas9"
         CDS             8405..8425
                         /label="NLS"
                         /note="nuclear localization signal of SV40 large T antigen"
         CDS             8426..8449
                         /label="DYKDDDDK tag"
         misc_feature    8488..8675
                         /label="SV40 polyA signal"
         promoter        8792..9032
                         /label="U6 Promoter"
                         /note="U6 promoter;RNA polymerase III promoter for human U6
                         snRNA"
    ORIGIN
            1 gtttaagagc tatgctggaa acagcatagc aagtttaaat aaggctagtc cgttatcaac
           61 ttgaaaaagt ggcaccgagt cggtgctttt tttgagcaaa aggccagcaa aaggccagga
          121 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
          181 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
          241 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
          301 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
          361 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
          421 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
          481 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
          541 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
          601 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
          661 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
          721 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
          781 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
          841 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
          901 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
          961 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
         1021 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
         1081 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
         1141 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
         1201 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
         1261 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
         1321 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
         1381 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
         1441 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
         1501 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
         1561 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
         1621 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
         1681 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
         1741 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
         1801 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
         1861 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcggcagt gaaaaaaatg
         1921 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
         1981 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
         2041 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcctctaga
         2101 ctgcagcctc aggagatctg ggcccctact tgtacagctc gtccatgccg tacaggaaca
         2161 ggtggtggcg gccctcggag cgctcgtact gttccacgat ggtgtagtcc tcgttgtggg
         2221 aggtgatgtc cagcttggtg tccacgtagt agtagccggg cagttgcacg ggcttcttgg
         2281 ccatgtagat ggtcttgaac tccaccaggt agtggccgcc gtccttcagc ttcagggcct
         2341 ggtggatctc gcccttcagc acgccgtcgc gggggtacag gcgctcggtg gaggcctccc
         2401 agcccatggt cttcttctgc attacggggc cgtcgggggg gaagttggtg ccgcgcatct
         2461 tcaccttgta gatcagcgtg ccgtcctgca gggaggagtc ctgggtcacg gtcaccagac
         2521 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttct
         2581 tgtaatcggg gatgtcggcg gggtgcttca cgtacgcctt ggagccgtac atgaactggg
         2641 gggacaggat gtcccaggcg aagggcaggg ggccgccctt ggtcaccttc agcttggcgg
         2701 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
         2761 tcatggagcc ctccatgcgc accttgaagc gcatgaactc tttgatgacg gccatgttgt
         2821 tgtcctcgga ggaggcggtg ccggagctgc cgctgccggt gctgccggtg ccatgcccca
         2881 ggaacaggtg gtggcggccc tcggagcgct cgtactgttc cacgatggtg tagtcctcgt
         2941 tgtgggaggt gatgtccagc ttggtgtcca cgtagtagta gccgggcagt tgcacgggct
         3001 tcttggccat gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc ttcagcttca
         3061 gggcctggtg gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc tcggtggagg
         3121 cctcccagcc catggtcttc ttctgcatta cggggccgtc gggggggaag ttggtgccgc
         3181 gcatcttcac cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg gtcacggtca
         3241 ccagaccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca
         3301 gcttcttgta atcggggatg tcggcggggt gcttcacgta cgccttggag ccgtacatga
         3361 actgggggga caggatgtcc caggcgaagg gcagggggcc gcccttggtc accttcagct
         3421 tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt
         3481 ggccgttcat ggagccctcc atgcgcacct tgaagcgcat gaactctttg atgacctcct
         3541 cgcccttgct caccatggtg gcgaattctc caggcgatct gacggttcac taaacgagct
         3601 ctgcttatat aggcctccca ccgtacacgc cacctcgaca tactcgagta gttattaata
         3661 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
         3721 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
         3781 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
         3841 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
         3901 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
         3961 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
         4021 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
         4081 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
         4141 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
         4201 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagggatcct ctagtcagct
         4261 gacgcgtgct agcgcggccg ccaccatgga caagaagtac agcatcggcc tggacatcgg
         4321 taccaacagc gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaagtt
         4381 caaggtgctg ggcaacaccg accgccacag catcaagaag aacctgatcg gcgccctgct
         4441 gttcgacagc ggcgagaccg ccgaggccac ccgcctgaag cgcaccgccc gccgccgcta
         4501 cacccgccgc aagaaccgca tctgctacct gcaggagatc ttcagcaacg agatggccaa
         4561 ggtggacgac agcttcttcc accgcctgga ggagagcttc ctggtggagg aggacaagaa
         4621 gcacgagcgc caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
         4681 ccccaccatc taccacctgc gcaagaagct ggtggacagc accgacaagg ccgacctgcg
         4741 cctgatctac ctggccctgg cccacatgat caagttccgc ggccacttcc tgatcgaggg
         4801 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
         4861 caaccagctg ttcgaggaga accccatcaa cgccagcggc gtggacgcca aggccatcct
         4921 gagcgcccgc ctgagcaaga gccgccgcct ggagaacctg atcgcccagc tgcccggcga
         4981 gaagaagaac ggcctgttcg gcaacctgat cgccctgagc ctgggcctga cccccaactt
         5041 caagagcaac ttcgacctgg ccgaggacgc caagctgcag ctgagcaagg acacctacga
         5101 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgttcctggc
         5161 cgccaagaac ctgagcgacg ccatcctgct gagcgacatc ctgcgcgtga acaccgagat
         5221 caccaaggcc cccctgagcg ccagcatgat caagcgctac gacgagcacc accaggacct
         5281 gaccctgctg aaggccctgg tgcgccagca gctgcccgag aagtacaagg agatcttctt
         5341 cgaccagagc aagaacggct acgccggcta catcgacggc ggcgccagcc aggaggagtt
         5401 ctacaagttc atcaagccca tcctggagaa gatggacggc accgaggagc tgctggtgaa
         5461 gctgaaccgc gaggacctgc tgcgcaagca gcgcaccttc gacaacggca gcatccccca
         5521 ccagatccac ctgggcgagc tgcacgccat cctgcgccgc caggaggact tctacccctt
         5581 cctgaaggac aaccgcgaga agatcgagaa gatcctgacc ttccgcatcc cctactacgt
         5641 gggccccctg gcccgcggca acagccgctt cgcctggatg acccgcaaga gcgaggagac
         5701 catcaccccc tggaacttcg aggaggtggt ggacaagggc gccagcgccc agagcttcat
         5761 cgagcgcatg accaacttcg acaagaacct gcccaacgag aaggtgctgc ccaagcacag
         5821 cctgctgtac gagtacttca ccgtgtacaa cgagctgacc aaggtgaagt acgtgaccga
         5881 gggcatgcgc aagcccgcct tcctgagcgg cgagcagaag aaggccatcg tggacctgct
         5941 gttcaagacc aaccgcaagg tgaccgtgaa gcagctgaag gaggactact tcaagaagat
         6001 cgagtgcttc gacagcgtgg agatcagcgg cgtggaggac cgcttcaacg ccagcctggg
         6061 cacctaccac gacctgctga agatcatcaa ggacaaggac ttcctggaca acgaggagaa
         6121 cgaggacatc ctggaggaca tcgtgctgac cctgaccctg ttcgaggacc gcgagatgat
         6181 cgaggagcgc ctgaagacct acgcccacct gttcgacgac aaggtgatga agcagctgaa
         6241 gcgccgccgc tacaccggct ggggccgcct gagccgcaag cttatcaacg gcatccgcga
         6301 caagcagagc ggcaagacca tcctggactt cctgaagagc gacggcttcg ccaaccgcaa
         6361 cttcatgcag ctgatccacg acgacagcct gaccttcaag gaggacatcc agaaggccca
         6421 ggtgagcggc cagggcgaca gcctgcacga gcacatcgcc aacctggccg gcagccccgc
         6481 catcaagaag ggcatcctgc agaccgtgaa ggtggtggac gagctggtga aggtgatggg
         6541 ccgccacaag cccgagaaca tcgtgatcga gatggcccgc gagaaccaga ccacccagaa
         6601 gggccagaag aacagccgcg agcgcatgaa gcgcatcgag gagggcatca aggagctggg
         6661 cagccagatc ctgaaggagc accccgtgga gaacacccag ctgcagaacg agaagctgta
         6721 cctgtactac ctgcagaacg gccgcgacat gtacgtggac caggagctgg acatcaaccg
         6781 cctgagcgac tacgacgtgg accacatcgt gccccagagc ttcctgaagg acgacagcat
         6841 cgacaacaag gtgctgaccc gcagcgacaa gaaccgcggc aagagcgaca acgtgcccag
         6901 cgaggaggtg gtgaagaaga tgaagaacta ctggcgccag ctgctgaacg ccaagctgat
         6961 cacccagcgc aagttcgaca acctgaccaa ggccgagcgc ggcggcctga gcgagctgga
         7021 caaggccggc ttcatcaagc gccagctggt ggagacccgc cagatcacca agcacgtggc
         7081 ccagatcctg gacagccgca tgaacaccaa gtacgacgag aacgacaagc tgatccgcga
         7141 ggtgaaggtg atcaccctga agagcaagct ggtgagcgac ttccgcaagg acttccagtt
         7201 ctacaaggtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
         7261 ggtgggcacc gccctgatca agaagtaccc caagctggag agcgagttcg tgtacggcga
         7321 ctacaaggtg tacgacgtgc gcaagatgat cgccaagagc gagcaggaga tcggcaaggc
         7381 caccgccaag tacttcttct acagcaacat catgaacttc ttcaagaccg agatcaccct
         7441 ggccaacggc gagatccgca agcgccccct gatcgagacc aacggcgaga ccggcgagat
         7501 cgtgtgggac aagggccgcg acttcgccac cgtgcgcaag gtgctgagca tgccccaggt
         7561 gaacatcgtg aagaagaccg aggtgcagac cggcggcttc agcaaggaga gcatcctgcc
         7621 caagcgcaac agcgacaagc tgatcgcccg caagaaggac tgggacccca agaagtacgg
         7681 cggcttcgac agccccaccg tggcctacag cgtgctggtg gtggccaagg tggagaaggg
         7741 caagagcaag aagctgaaga gcgtgaagga gctgctgggc atcaccatca tggagcgcag
         7801 cagcttcgag aagaacccca tcgacttcct ggaggccaag ggctacaagg aggtgaagaa
         7861 ggacctgatc atcaagctgc ccaagtacag cctgttcgag ctggagaacg gccgcaagcg
         7921 catgctggcc agcgccggcg agctgcagaa gggcaacgag ctggccctgc ccagcaagta
         7981 cgtgaacttc ctgtacctgg ccagccacta cgagaagctg aagggcagcc ccgaggacaa
         8041 cgagcagaag cagctgttcg tggagcagca caagcactac ctggacgaga tcatcgagca
         8101 gatcagcgag ttcagcaagc gcgtgatcct ggccgacgcc aacctggaca aggtgctgag
         8161 cgcctacaac aagcaccgcg acaagcccat ccgcgagcag gccgagaaca tcatccacct
         8221 gttcaccctg accaacctgg gcgcccccgc cgccttcaag tacttcgaca ccaccatcga
         8281 ccgcaagcgc tacaccagca ccaaggaggt gctggacgcc accctgatcc accagagcat
         8341 caccggtctg tacgagaccc gcatcgacct gagccagctg ggcggcgacg gcggctccgg
         8401 acctccaaag aaaaagagaa aagtagacta caaggacgac gatgacaagt aaatcgataa
         8461 gcttgtcgac gatatctcca gaggatcata atcagccata ccacatttgt agaggtttta
         8521 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
         8581 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
         8641 aatttcacaa ataaagcatt tttttcactg ccccgagctt cctcgctcac tgactcgctg
         8701 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
         8761 tccacagaat caggggataa cgcaggaaag agagggccta tttcccatga ttccttcata
         8821 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac
         8881 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
         8941 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
         9001 tttcttggct ttatatatct tgtggaaagg acgaaacacc gg
    //
    
    

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