pIRESpuro3

价格:1200元

联系方式:I47-825O-882O

买家导航

载体基本信息

出品公司: Clontech
载体名称: pIRESpuro3.html' target='_blank'>pIRESpuro3
质粒类型: 哺乳动物细胞表达载体
启动子: CMV
表达水平:
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5157 bp
5' 测序引物:   CMV-F: 5'-CGCAAATGGGCGGTAGGCGTG-3'
3' 测序引物: pIRES-R:5'-GCCCTAGATGCATGCTCG-3'
载体标签:
载体抗性: 氨苄青霉素(Ampicillin)
筛选标记: 嘌呤霉素(Puromycin)
克隆菌株: DH5α, HB101
宿主细胞(系): 常规细胞系,293、CV-1、CHO等
备注:
pIRESpuro3载体含有IRES元件,目的基因和抗性基因以双顺反子的形式表达。
产品目录号: 6986-1
稳定性: 稳表达 或 稳表达
组成型: 组成型 
病毒/非病毒: 非病毒

载体质粒图谱和多克隆位点信息

pIRESpuro3载体图谱和多克隆位点

载体简介

Description
pIRESpuro3.html' target='_blank'>pIRESpuro3 contains the internal ribosome entry site (IRES) of the encephalomyocarditis virus (ECMV), which permits the translation of two open reading frames from one messenger RNA (1–3). After selection with puromycin, nearly all surviving colonies will stably express the gene of interest, thus decreasing the need to screen large numbers of colonies to find functional clones. To select for cells that express high levels of the gene of interest, the selective pressure for antibiotic resistance was increased by shifting the puromycin resistance gene downstream to a less optimal position for translation as directed by the IRES sequence (3). By decreasing the level of expression of the antibiotic resistance marker, the selective pressure on the entire expression cassette is increased, resulting in selection for cells that express the entire transcript, including the gene of interest, at high levels. The expression cassette of pIRESpuro3 contains the human cytomegalovirus (CMV) major immediate early promoter/enhancer followed by a multiple cloning site (MCS) that precedes stop codons in all three reading frames, a synthetic intron known to enhance the stability of the mRNA (4), the ECMV IRES followed by the gene encoding puromycin resistance (puromycin-N-acetyltransferase; 5), and the polyadenylation signal from SV40. Ribosomes can enter the bicistronic mRNA at the 5' end to translate the gene of interest and at the ECMV IRES to translate the antibiotic resistance marker.  

Use
When using the pIRESpuro3 Vector, the antibiotic exerts selective pressure on the whole expression cassette; thus, a high dose of antibiotic will select for cells expressing a high level of the gene of interest. This selective pressure also ensures that the expression of the gene of interest will be stable  

载体序列

LOCUS       pIRESpuro3.html' target='_blank'>pIRESpuro3   5171 bp    DNA   circular            16-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.biofeng.com
COMMENT     Cat. No. 631619
FEATURES             Location/Qualifiers
     misc_feature    309..810
                     /label=CMV\IE\Promoter/Enhancer
     misc_feature    863..879
                     /label=T7\RNA\Polymerase\Promoter
     misc_feature    912..1015
                     /label=MCS
     misc_feature    1015..1310
                     /label=IVS
     misc_feature    1336..1926
                     /label=IRES
     misc_feature    1959..2555
                     /label=Puro(R)
     misc_feature    2822..2827
                     /label=SV40\polyA\signal
     misc_feature    2851..2856
                     /label=SV40\polyA\signal
     misc_feature    3411..4010
                     /label=pUC\origin
     misc_feature    complement(4172..5032)
                     /label=Amp(R)
BASE COUNT     1207 a      1429 c      1303 g      1232 t 
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc 
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtcgtacgt 
      961 ctccggattc gaattcggat ccgcggccgc atagataact gatccagtgt gctggaatta 
     1021 attcgctgtc tgcgagggcc agctgttggg gtgagtactc cctctcaaaa gcgggcatga 
     1081 cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc acctggcccg 
     1141 cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga aaagacaatc tttttgttgt 
     1201 caagcttgag gtgtggcagg cttgagatct ggccatacac ttgagtgaca atgacatcca 
     1261 ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcaggtcg agcatgcatc 
     1321 tagggcggcc aattccgccc ctctcccccc cccccctctc cctccccccc ccctaacgtt 
     1381 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc 
     1441 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc 
     1501 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag 
     1561 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg 
     1621 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat 
     1681 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga 
     1741 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc 
     1801 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg 
     1861 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga 
     1921 tgataagctt gccacaaccc acaaggagac gaccttccat gaccgagtac aagcccacgg 
     1981 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg 
     2041 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg 
     2101 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg 
     2161 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt 
     2221 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac 
     2281 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg 
     2341 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag 
     2401 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc 
     2461 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac 
     2521 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc 
     2581 gcccgaccga aaggagcgca cgaccccatg gctccgaccg aagccacccg gggcggcccc 
     2641 gccgaccccg cacccgcccc cgaggcccac cgactctaga taactgatca taatcagcca 
     2701 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 
     2761 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta 
     2821 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 
     2881 ttgtggtttg tccaaactca tcaatgtatc ttaacgctcg agtgcattct agttgtggtt 
     2941 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 
     3001 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 
     3061 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 
     3121 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 
     3181 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 
     3241 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 
     3301 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 
     3361 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 
     3421 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 
     3481 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 
     3541 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 
     3601 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 
     3661 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 
     3721 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 
     3781 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 
     3841 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 
     3901 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 
     3961 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 
     4021 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 
     4081 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 
     4141 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 
     4201 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 
     4261 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 
     4321 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 
     4381 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 
     4441 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 
     4501 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 
     4561 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 
     4621 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 
     4681 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 
     4741 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 
     4801 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 
     4861 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 
     4921 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 
     4981 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 
     5041 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 
     5101 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 
     5161 cacctgacgt c 
//

载体质粒图谱pdf版和相关资料下载

载体应用举例

分享到:

全部载体分类

相关载体

相关文章

相关问答

Copyright © 20012-2013 BIOFENG. 生物风 版权所有 Powered by Biofeng