| 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 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
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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