pRetroX-Tight-Pur-Luc

价格:4000元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Clontech
载体名称: pRetroX-Tight-Pur-Luc
质粒类型: 逆转录病毒载体;四环素诱导载体;荧光素酶报告载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: Tight
载体大小: 8201 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: 氨苄青霉素
筛选标记: 嘌呤霉素(Puromycin)
克隆菌株: DH5α 等通用菌株
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 逆转录病毒载体pRetroX-Tight-Pur-Luc含有荧光素酶报告基因, 是pRetroX-Tight-Pur载体的阳性对照载体。
产品目录号: 632104
稳定性: 稳表达
组成型/诱导型: 组成型
病毒/非病毒: 逆转录病毒

载体质粒图谱和多克隆位点信息

pRetroX-Tight-Pur-Luc载体图谱



pRetroX-Tight-Pur-Luc 载体特征

载体简介

pRetroX-Tight-Pur-Lu载体描述

pRetroX-Tight-Pur-Luc is a retroviral control vector that expresses firefly luciferase under the control of PTight, a modified 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 modified minimal CMV promoter (1). This vector is supplied with our Retro-X?Tet-On? and Tet-Off? Advanced Inducible Expression Systems (Cat. Nos. 632104 and 632105). 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-Pur-Luc is optimized to eliminate promoter interference through LTR self-inactivation. 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, resulting in the inactivation of the 5’ LTR CMV enhancer sequence. This mechanism can reduce the phenomenon known as promoter interference (9-10) and allow more efficient 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-Pur-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-Pur-Luc contains a puromycin resistance gene (Purr) 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-Pur-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?, DH10B?, 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 vector sequence was 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 sequenced.
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.

References
1. pTRE-Tight Vectors (April 2003) Clontechniques XVIII(3):13-14.
2. Gossen, M. & Bujard, H. (1992) Proc. Natl. Acad. Sci USA 89(12):5547–5551.
3. Gossen, M., et al. (1995) Science 268(5218):1766–1769.
4. Urlinger, S. et al. (2000) Proc. Natl. Acad. Sci. USA 97(14):7963-7968.
5. Inducible Gene Expression Systems (January 2007) Clontechniques XXII(1):1-2.
6. Tet-On Advanced Inducible Gene Expression System (2006) Clontechniques XXI(2):1-3.
7. Ory, D.S. et al. (1996) Proc. Natl. Acad. Sci. USA 93(21):11400-11406.
8. Pear, W.S. et al. (1993) Proc. Natl. Acad. Sci. USA 90(18):8392-8396.
9. Barton, G.M. & Medzhitov R. (2002) Proc. Natl. Sci. USA 99(23):14943-14945.
10. Emerman, M. & Temin, H.M. (1984) Cell 39(3 pt. 2):449-467.

载体序列

LOCUS       pRetroX-Tight-Pur-Luc   8200 bp    DNA   circular            10-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631128,632104,631129,632105
FEATURES             Location/Qualifiers
     misc_feature    1..727
                     /label=5'\LTR\(CMV/MSV)
     misc_feature    758..1567
                     /label=Extended\packaging\signal
     misc_feature    1602..1917
                     /label=Tight\Promoter
     misc_feature    1934..3586
                     /label=Luciferase
     misc_feature    3602..4105
                     /label=PGK\Promoter
     misc_feature    4126..4725
                     /label=Puro(R)
     misc_feature    4946..5331
                     /label=3'\MMLV\LTR\(del)
     misc_feature    5611..5878
                     /label=SV40\Promoter
     misc_feature    5832..5897
                     /label=SV40\origin
     misc_feature    6219..6219
                     /label=pUC\origin
     misc_feature    complement(6978..7838)
                     /label=Amp(R)
BASE COUNT     1943 a      2196 c      2099 g      1962 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 cagcccaagc ttaccatgac cgagtacaag 
     4141 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc 
     4201 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg 
     4261 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg 
     4321 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg 
     4381 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg 
     4441 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg 
     4501 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc 
     4561 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc 
     4621 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc 
     4681 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac 
     4741 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cggcgtctcg agatatcagt 
     4801 ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc 
     4861 atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac 
     4921 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata 
     4981 actgagaata gagaagttca gatcaaggtc aggaacagat ggaacagggt cgaccctaga 
     5041 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga 
     5101 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata 
     5161 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt 
     5221 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt 
     5281 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc 
     5341 gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg 
     5401 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 
     5461 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 
     5521 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta 
     5581 gatccggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 
     5641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 
     5701 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 
     5761 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 
     5821 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 
     5881 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttactggctt 
     5941 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg 
     6001 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac 
     6061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg 
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 
     6901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 
     6961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 
     7021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 
     7081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 
     7141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 
     7201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 
     7261 gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg 
     7321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 
     7381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 
     7441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 
     7501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 
     7561 atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc 
     7621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 
     7681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 
     7741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 
     7801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 
     7861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 
     7921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa 
     7981 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt 
     8041 caagaattag cttggccatt gcatacgttg tatccatatc ataatatgta catttatatt 
     8101 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa 
     8161 tcaattacgg ggtcattagt tcatagccca tatatggagt 
//

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