pRetroX-Tight-Hyg-Luc

价格:3500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-Hyg-Luc
质粒类型: 逆病毒载体;四环素调控载体;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: 9054 bp
5' 测序引物及序列: 5’-ATCTGAGGCCCTTTCGTCTTCACT-3’
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: 潮霉素(Hygromycin)
克隆菌株: DH5α
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 逆病毒载体pRetroX-Tight-Hyg-Luc是四环素诱导载体,作为pRetroX-Tight-Hyg载体的阳性对照。
产品目录号: 631034
稳定性: 稳表达
组成型/诱导型: 诱导型
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-Hyg-Luc载体图谱



pRetroX-Tight-Hyg-Luc 载体特征

载体简介

pRetroX-Tight-Hyg-Luc载体描述

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.

载体序列

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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