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基本信息
载体类型: | 哺乳动物表达载体;双顺反子表达载体 |
载体抗性: | Amp |
载体标签: | EGFP |
启动子: | Cag |
筛选标记: | Neo |
表达水平: | 高拷贝 |
载体大小: | 6183bp |
克隆方法: | 多克隆位点,限制性内切酶 |
5' 测序引物: | pCAG-F: GCAACGTGCTGGTTATTGTG |
3' 测序引物: | IRES-R : CCTCACATTGCCAAAAGACG |
备注: | 如有侵权请产权持有人尽快联系法务15927229196 |
质粒图谱和多克隆位点信息
注意:
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载体描述
pGAG-IRES-GFP在pGAG-GFP载体的基础上改造而来,双顺反子载体;荧光报告载体。
CAG启动子是人工构建的组合启动子,由巨细胞病毒(the cytomegalovirus,CMV)早期增强子(early enhancer element)和鸡β-肌动蛋白(chicken beta-actin)启动子组成,用于驱动基因在哺乳动物载体的高水平表达。真核表达的启动子CAG(CMV/actin/Globin),它减少了单纯CMV启动子在多种细胞中容易被甲基化从而被关闭表达的尴尬。
pGAG-IRES-GFP contains the internal ribosome entry site (IRES; 1, 2) of the encephalomyocarditis virus (ECMV) between the MCS and the enhanced green fluorescent protein (EGFP) coding region. This permits both the gene of interest (cloned into the MCS) and the EGFP gene to be translated from a single bicistronic mRNA.
载体序列
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg cgcgccatcg
1741 atacgcgtca attgcggccg cctcgagctc aagcttctag ttctgcagtc gacggtaccg
1801 cgggcccggg atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt
1861 ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg
1921 caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc
1981 ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga
2041 agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc
2101 tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc
2161 acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc
2221 aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga
2281 tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg
2341 ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa
2401 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
2461 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
2521 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
2581 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
2641 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
2701 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
2761 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
2821 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
2881 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
2941 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
3001 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
3061 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
3121 aagaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
3181 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
3241 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
3301 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
3361 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
3421 tgaacaaagg tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
3481 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
3541 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
3601 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatgaagat ccctcgacct
3661 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
3721 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
3781 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
3841 tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
3901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
3961 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
4021 aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat tgcagcttat
4081 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
4141 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgctgc
4201 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
4261 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
4321 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
4381 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
4441 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
4501 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
4561 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
4621 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
4681 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
4741 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
4801 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
4861 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
4921 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
4981 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
5041 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
5101 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
5161 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
5221 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
5281 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
5341 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
5401 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
5461 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
5521 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
5581 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
5641 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
5701 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
5761 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
5821 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
5881 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
5941 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
6001 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
6061 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
6121 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
6181 ctg
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