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          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



          pRetroX-Tight-Hyg-Luc 載體特征


          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
          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.


          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
               misc_feature    4123..5148
               LTR             5748..6184
               CDS             complement(7830..8690)
               promoter        1602..1917
               LTR             1..727
               misc_feature    758..1567
               promoter        6463..6730
               rep_origin      6684..6749
               rep_origin      7071..7071
               misc_feature    1934..3586
          BASE COUNT     2184 a      2335 c      2332 g      2201 t 
                  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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