pRetro-Lib

价格:3000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: pRetro-Lib
质粒类型: 逆转录病毒载体;文库克隆与表达载体
高拷贝/低拷贝: 低拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: --
载体大小: 4891 bp
5' 测序引物及序列: 5'-AGCCCTCACTCCTTCTCTAG-3'
3' 测序引物及序列: 5'-ACCTACAGGTGGGGTCTTTCATTCCC-3'
载体标签: --
载体抗性: 氨苄青霉素
筛选标记: --
克隆菌株: DH5α 等
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: --
产品目录号: 635002
稳定性: 稳表达 或 瞬表达
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

pRetro-Lib载体图谱



pRetro-Lib 多克隆位点

pRetro-Lib 载体特征

载体简介

pRetro-Lib载体描述

pRetro-Lib, derived from Moloney murine leukemia virus (MoMuLV), is designed for delivery and expression of retroviral libraries or genes (1–3). Upon transfection into a packaging cell line, pRetro-Lib can transiently express, or integrate and stably express, a transcript containing the gene of interest. The 5' viral LTR in this vector contains a viral promoter that controls expression of genes cloned into the MCS. pRetro-Lib also contains the Col E1 origin of replication and E. coli Ampr gene for propagation and antibiotic selection in bacteria. pRetro-Lib can be used as an alternative vector in the construction of a cDNA library using the SMART? cDNA Library Construction Kit (Cat. No. 634901). Retroviral expression libraries can be constructed by directionally cloning the SfiI(dT)30 primed cDNA into the SfiIA/B sites of pRetro-Lib. Restriction digestion with SfiI will remove the Stuffer fragment, prior to cloning.

pRetro-Lib facilitates expression cloning in combination with the benefits of retroviral gene delivery. Upon transfection into a high-titer packaging cell line, RNA from the vector is packaged into infectious, replication-incompetent retroviral particles. That is, these retroviral particles can infect target cells and transmit the gene of interest (which is cloned between the viral LTR sequences), but cannot replicate within these cells since the cells lack the viral structural genes. pRetro-Lib does not contain the structural genes necessary for particle formation and replication, gag, pol, and env; these genes are stably integrated into the packaging line. Subsequent introduction of pRetro-Lib, containing Ψ+, transcription and processing elements, and the gene of interest produces high-titer, replication-incompetent infectious virus.

Propagation in E. coli:
 Suitable host strains: DH5α, and other general purpose strains.
 Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) to E. coli hosts.
 E. coli replication origin: Col E1
 Copy number: low
 Plasmid incompatibility group: pBR322

NOTE: The viral supernatants produced by this retroviral vector could, depending on your cloned insert, 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. Coffin, J. M. & Varmus, H. E., Eds. (1996) Retroviruses (Cold Spring Harbor Laboratory Press, NY).
2. Ausubel, F. M., et al. (1994) Current Protocols in Molecular Biology (Greene Publishing Associates, Inc. & John Wiley & Sons, Inc.).
3. Miller, A. D. & Rosman, G. J. (1989) BioTechniques 7:980–990.


载体序列

LOCUS       pRetro-Lib   4891 bp    DNA   circular            20-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 635002
FEATURES             Location/Qualifiers
     misc_feature    1..589
                     /label=5'\LTR
     misc_feature    659..1468
                     /label=Extended\packaging\signal
     misc_feature    1470..1502
                     /label=MCS-A
     misc_feature    1503..1694
                     /label=Stuffer\Fragment
     misc_feature    1695..1723
                     /label=MCS-B
     misc_feature    1763..2356
                     /label=3'\LTR
     misc_feature    2892..2892
                     /label=pUC\origin
     misc_feature    complement(3652..4512)
                     /label=Amp(R)
BASE COUNT     1184 a      1325 c      1187 g      1195 t 
ORIGIN
        1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat 
       61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc 
      121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca 
      181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg 
      241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa 
      301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac 
      361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa 
      421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac 
      481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg 
      541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt 
      601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc 
      661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg 
      721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt 
      781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg 
      841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt 
      901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa 
      961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct 
     1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt 
     1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa 
     1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc 
     1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc 
     1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt 
     1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc 
     1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta 
     1441 tccagccctc actccttctc taggcgccgg aattcgttag gccattacgg cctgcaggat 
     1501 ccgaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 
     1561 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 
     1621 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 
     1681 atcttatcat gtctggatcc ggccgcctcg gccaaacatc gataaaataa aagattttat 
     1741 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt 
     1801 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat 
     1861 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca 
     1921 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga 
     1981 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc 
     2041 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc 
     2101 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc 
     2161 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc 
     2221 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat 
     2281 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca 
     2341 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc 
     2401 gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc 
     2461 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca 
     2521 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc 
     2581 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt 
     2641 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg 
     2701 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 
     2761 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 
     2821 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 
     2881 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 
     2941 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 
     3001 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 
     3061 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 
     3121 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 
     3181 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 
     3241 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 
     3301 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 
     3361 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 
     3421 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 
     3481 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 
     3541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 
     3601 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 
     3661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 
     3721 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 
     3781 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 
     3841 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 
     3901 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 
     3961 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 
     4021 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 
     4081 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 
     4141 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 
     4201 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 
     4261 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 
     4321 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 
     4381 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 
     4441 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 
     4501 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 
     4561 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 
     4621 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta 
     4681 taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttgctagcaa ttgctagcaa 
     4741 ttgctagcaa ttcataccag atcaccgaaa actgtcctcc aaatgtgtcc ccctcacact 
     4801 cccaaattcg cgggcttctg cctcttagac cactctaccc tattccccac actcaccgga 
     4861 gccaaagccg cggcccttcc gtttctttgc t 
//

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