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  • pRetroX-Tight-Hyg-Luc

    價格:3500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: pRetroX-Tight-Hyg-Luc
    質粒類型: 逆病毒載體;四環素調控載體;熒光素酶報告載體
    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: Tight
    載體大小: 9054 bp
    5' 測序引物及序列: 5’-ATCTGAGGCCCTTTCGTCTTCACT-3’
    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: 潮霉素(Hygromycin)
    克隆菌株: DH5α
    宿主細胞(系): 常規細胞系(293、CV-1、CHO等)
    備注: 逆病毒載體pRetroX-Tight-Hyg-Luc是四環素誘導載體,作為pRetroX-Tight-Hyg載體的陽性對照。
    產品目錄號: 631034
    穩定性: 穩表達
    組成型/誘導型: 誘導型
    病毒/非病毒: 逆轉錄病毒

    載體質粒圖譜和多克隆位點信息

    pRetroX-Tight-Hyg-Luc載體圖譜



    pRetroX-Tight-Hyg-Luc 載體特征

    載體簡介

    pRetroX-Tight-Hyg-Luc載體描述
    
    pRetroX-Tight-Hyg-Luc is a retroviral control vector (supplied with pRetroX-Tight-Hyg) that expresses fi refl y luciferase under the control of PTight, a modifi ed Tet-responsive promoter. PTight consists of seven direct repeats of a 36 bp regulatory sequence that contains the 19 bp tet operator sequence (tetO), and a modifi ed minimal CMV promoter (1). This vector is designed to be used as a control vector with Clontech's Advanced Inducible Expression Systems. These systems provide ready access to the inducible gene expression strategy of Gossen & Bujard, and incorporate major improvements described by Urlinger, et al. (2-6).
    
    pRetroX-Tight-Hyg-Luc is optimized to eliminate promoter interference through LTR selfinactivation. The hybrid 5’ LTR consists of the cytomegalovirus (CMV) type I enhancer and the mouse sarcoma virus (MSV) promoter. This promoter drives high levels of viral genome transcription in HEK 293-based packaging cell lines due, in part, to the presence of adenoviral E1A (7-8) in these cells. The self-inactivating feature of the vector is provided by a deletion in the 3’ LTR enhancer region (U3). During reverse transcription of the retroviral RNA, a copy of the inactivated 3’ LTR U3 region replaces the corresponding region of the 5’ LTR, resultingin the inactivation of the 5’ LTR CMV enhancer sequence. This mechanism can reduce the phenomenon known as promoter interference (9-10) and allow more effi cient expression. In
    this way, pRetroX-Tight-Hyg-Luc supports high viral titers, yet eliminates potential downstream interference between the inducible PTight promoter and its adjacent viral LTR.
    
    pRetroX-Tight-Hyg-Luc contains all of the necessary viral RNA processing elements; these include the 5’ and 3’ LTRs, the packaging signal (Ψ+), and the tRNA primer binding site. For safety reasons, however, the vector lacks the structural genes (gag, pol, and env) necessary for retroviral particle formation and replication. pRetroX-Tight-Hyg-Luc contains a hygromycin resistance gene (Hygr) under the control of the murine phosphoglycerate kinase (PGK) promoter (PPGK) for the selection of stable transfectants. In addition, the vector contains a ColE1 origin of replication and an E. coli Ampr gene for propagation and selection in bacteria.
    
    pRetroX-Tight-Hyg-Luc can be used as a control vector for either plasmid or retroviral expression vectors. When used as a control for retroviral expression vectors, it must be transfected into a packaging cell line, such as GP2-293; we recommend the Retro-X Universal Packaging System (Cat. No. 631530). Packaging cell lines allow you to produce infectious, replication-incompetent retroviral particles. These retroviral particles can infect a wide range of target cells, but they cannot replicate within these cells due to the absence of viral structural genes. The separate introduction and integration of the structural genes into the packaging cell line minimizes the chance of producing replication-competent virus due to recombination events during cell proliferation.
    
    Propagation in E. coli
     Suitable host strains: DH5α and other general purpose strains.
     Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
     E. coli replication origin: ColE1
     Copy number: high
    
    Notes:
    The attached sequence fi le has been compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequence.
    The viral supernatants produced by this retroviral vector could contain potentially hazardous recombinant virus. Due caution must be exercised in the production and handling of recombinant retrovirus. Appropriate NIH, regional, and institutional guidelines apply.
    
    

    載體序列

    LOCUS       pRetroX-Tight-Hyg-Luc   9052 bp    DNA   circular            10-NOV-2009
    COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
    COMMENT     Cat. No. 631034
    FEATURES             Location/Qualifiers
         promoter        3602..4105
                         /label=PGK\Promoter
         misc_feature    4123..5148
                         /label=Hyg(R)
         LTR             5748..6184
                         /label=3'\MMLV\LTR\(del)
         CDS             complement(7830..8690)
                         /label=Amp(R)
         promoter        1602..1917
                         /label=Tight\Promoter
         LTR             1..727
                         /label=5'\LTR\(CMV/MSV)
         misc_feature    758..1567
                         /label=Packaging\Signal
         promoter        6463..6730
                         /label=SV40\Promoter
         rep_origin      6684..6749
                         /label=SV40\origin
         rep_origin      7071..7071
                         /label=pUC\origin
         misc_feature    1934..3586
                         /label=Luciferase
    BASE COUNT     2184 a      2335 c      2332 g      2201 t 
    ORIGIN
            1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc 
           61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac 
          121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata 
          181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc 
          241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta 
          301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac 
          361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc 
          421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc 
          481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc 
          541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc 
          601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat 
          661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc 
          721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc 
          781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta 
          841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc 
          901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc 
          961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca 
         1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct 
         1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc 
         1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga 
         1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt 
         1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg 
         1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga 
         1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa 
         1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct 
         1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct 
         1561 aggcgccgga attgaagatc tgaggccctt tcgtcttcac tcgagtttac tccctatcag 
         1621 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgat gtcgagttta 
         1681 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt 
         1741 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttatc cctatcagtg 
         1801 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgaggtagg 
         1861 cgtgtacggt gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg 
         1921 agaaggatcc accatggaag acgccaaaaa cataaagaaa ggcccggcgc cattctatcc 
         1981 gctagaggat ggaaccgctg gagagcaact gcataaggct atgaagagat acgccctggt 
         2041 tcctggaaca attgctttta cagatgcaca tatcgaggtg aacatcacgt acgcggaata 
         2101 cttcgaaatg tccgttcggt tggcagaagc tatgaaacga tatgggctga atacaaatca 
         2161 cagaatcgtc gtatgcagtg aaaactctct tcaattcttt atgccggtgt tgggcgcgtt 
         2221 atttatcgga gttgcagttg cgcccgcgaa cgacatttat aatgaacgtg aattgctcaa 
         2281 cagtatgaac atttcgcagc ctaccgtagt gtttgtttcc aaaaaggggt tgcaaaaaat 
         2341 tttgaacgtg caaaaaaaat taccaataat ccagaaaatt attatcatgg attctaaaac 
         2401 ggattaccag ggatttcagt cgatgtacac gttcgtcaca tctcatctac ctcccggttt 
         2461 taatgaatac gattttgtac cagagtcctt tgatcgtgac aaaacaattg cactgataat 
         2521 gaactcctct ggatctactg ggttacctaa gggtgtggcc cttccgcata gaactgcctg 
         2581 cgtcagattc tcgcatgcca gagatcctat ttttggcaat caaatcattc cggatactgc 
         2641 gattttaagt gttgttccat tccatcacgg ttttggaatg tttactacac tcggatattt 
         2701 gatatgtgga tttcgagtcg tcttaatgta tagatttgaa gaagagctgt ttttacgatc 
         2761 ccttcaggat tacaaaattc aaagtgcgtt gctagtacca accctatttt cattcttcgc 
         2821 caaaagcact ctgattgaca aatacgattt atctaattta cacgaaattg cttctggggg 
         2881 cgcacctctt tcgaaagaag tcggggaagc ggttgcaaaa cgcttccatc ttccagggat 
         2941 acgacaagga tatgggctca ctgagactac atcagctatt ctgattacac ccgaggggga 
         3001 tgataaaccg ggcgcggtcg gtaaagttgt tccatttttt gaagcgaagg ttgtggatct 
         3061 ggataccggg aaaacgctgg gcgttaatca gagaggcgaa ttatgtgtca gaggacctat 
         3121 gattatgtcc ggttatgtaa acaatccgga agcgaccaac gccttgattg acaaggatgg 
         3181 atggctacat tctggagaca tagcttactg ggacgaagac gaacacttct tcatagttga 
         3241 ccgcttgaag tctttaatta aatacaaagg ataccaggtg gcccccgctg aattggagtc 
         3301 gatattgtta caacacccca acatcttcga cgcgggcgtg gcaggtcttc ccgacgatga 
         3361 cgccggtgaa cttcccgccg ccgttgttgt tttggagcac ggaaagacga tgacggaaaa 
         3421 agagatcgtg gattacgtcg ccagtcaagt aacaaccgcg aaaaagttgc gcggaggagt 
         3481 tgtgtttgtg gacgaagtac cgaaaggtct taccggaaaa ctcgacgcaa gaaaaatcag 
         3541 agagatcctc ataaaggcca agaagggcgg aaagtccaaa ttgtaaaata cgcgtgaatt 
         3601 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc 
         3661 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg 
         3721 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct 
         3781 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt 
         3841 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg 
         3901 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg 
         3961 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag 
         4021 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc 
         4081 ctcttcctca tctccgggcc tttcgacctg catcccgcca ccatgaaaaa gcctgaactc 
         4141 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg 
         4201 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat 
         4261 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac 
         4321 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc 
         4381 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc 
         4441 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat 
         4501 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca 
         4561 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg 
         4621 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag 
         4681 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg 
         4741 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa 
         4801 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg 
         4861 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg 
         4921 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca 
         4981 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt 
         5041 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc 
         5101 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagag tagatgccga 
         5161 ccgaacaaga gctgatttcg agaacgcctc agccagcaac tcgcgcgagc ctagcaaggc 
         5221 aaatgcgaga gaacggcctt acgcttggtg gcacagttct cgtccacagt tcgctaagct 
         5281 cgctcggctg ggtcgcggga gggccggtcg cagtgattca ggcccttctg gattgtgttg 
         5341 gtccccaggg cacgattgtc atgcccacgc actcgggtga tctgactgat cccgcagatt 
         5401 ggagatcgcc gcccgtgcct gccgattggg tgcagatccg tcgacctgca gccaagctta 
         5461 tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc 
         5521 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac 
         5581 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa 
         5641 caggatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat 
         5701 agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat 
         5761 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg 
         5821 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagg 
         5881 gtcgacccta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg 
         5941 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc 
         6001 ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg 
         6061 agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc 
         6121 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc 
         6181 atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg 
         6241 gaggtaagct ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 
         6301 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 
         6361 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga 
         6421 tagcggagtg tagatccggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 
         6481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 
         6541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 
         6601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 
         6661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 
         6721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 
         6781 gcttactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 
         6841 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 
         6901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 
         6961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 
         7021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 
         7081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 
         7141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 
         7201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 
         7261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 
         7321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 
         7381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 
         7441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 
         7501 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 
         7561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 
         7621 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 
         7681 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 
         7741 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 
         7801 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 
         7861 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 
         7921 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 
         7981 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 
         8041 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 
         8101 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg 
         8161 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 
         8221 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 
         8281 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 
         8341 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 
         8401 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc 
         8461 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 
         8521 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 
         8581 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 
         8641 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 
         8701 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 
         8761 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 
         8821 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 
         8881 ccctttcgtc ttcaagaatt agcttggcca ttgcatacgt tgtatccata tcataatatg 
         8941 tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt 
         9001 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gt 
    //
    

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