| 出品公司: | Clontech |
|---|---|
| 载体名称: | pIRES |
| 质粒类型: | 哺乳动物细胞表达载体 |
| 启动子: | CMV |
| 表达水平: | 高 |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 6092 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: | Ampicillin |
| 筛选标记: | Neomycin/G418 |
| 备注: | Express 2 genes on one transcript with IRES. |
| 产品目录号: | 631605 |
| 稳定性: | 稳表达 Stable |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
pIRES is a mammalian expression vector that allows high level expression of two genes of interest from the same bicistronic mRNA transcript. The vector contains the encephalomyocarditis virus (ECMV) internal ribosome entry site (IRES) flanked by two multiple cloning sites (MCS A and B), an arrangement that allows cap-independent translation of the gene cloned into MCS B (1–3). pIRES utilizes a partially disabled IRES sequence (1) that reduces the rate at which the gene cloned into MCS B is translated relative to that of MCS A.
Expression of the bicistronic transcript is driven by the constitutively active cytomegalovirus immediate early promoter (PCMV IE), located upstream of MCS A. An intervening sequence (IVS) known to enhance the stability of mRNA (4) is located between PCMV IE and MCS A, and is efficiently spliced out following transcription. SV40 polyadenylation signals downstream of MCS B direct proper processing of the 3' end of the mRNA. Bacteriophage T7 and T3 promoters are located upstream of MCS A and downstream of MCS B, respectively. pIRES includes a neomycin resistance gene (Neor) to aid in the selection of transfected cells. Neor is expressed from the SV40 enhancer/promoter, and a synthetic polyadenyla¬tion signal directs proper processing of the 3' end of the Neor mRNA. The SV40 origin al¬lows for replication in mammalian cells expressing the SV40 T antigen. The vector also contains an ampicillin resistance gene (Ampr), and a ColE1 origin of replication for se¬lection and propagation in E. coli, and an f1 origin for single-stranded DNA production.
载体序列
LOCUS pIRES 6092 bp DNA circular SYN
DEFINITION pIRES
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biovector.net/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biovector.net|
FEATURES Location/Qualifiers
source 1..6092
/organism="pIRES"
/mol_type="other DNA"
promoter 163..723
/label="CMV_immearly_promoter"
misc_feature 218..505
/label="CAG_enhancer"
promoter 669..750
/label="CMV_promoter"
misc_feature 680..700
/label="CMV_fwd_primer"
misc_feature 724..743
/label="pCEP_fwd_primer"
misc_feature 726..750
/label="LNCX_primer"
misc_feature 890..1022
/label="chim_int"
promoter 1067..1085
/label="T7_promoter"
misc_feature 1136..1699
/label="IRES"
terminator complement(1791..1910)
/label="SV40_PA_terminator"
misc_feature complement(1805..1824)
/label="EBV_rev_primer"
rep_origin 2118..2424
/label="f1_origin"
misc_feature complement(2698..2718)
/label="pBABE_3_primer"
misc_feature complement(2704..2919)
/label="SV40_enhancer"
promoter 2716..2984
/label="SV40_promoter"
rep_origin 2883..2960
/label="SV40_origin"
misc_feature 2945..2964
/label="SV40pro_F_primer"
CDS 3083..3877
/label="ORF frame 2"
gene 3086..3874
/label="NeoR/KanR"
/gene="NeoR/KanR"
CDS complement(3392..3928)
/label="ORF frame 2"
terminator 3941..3989
/label="synth_PA_terminator"
misc_feature 4149..4171
/label="pGEX_3_primer"
promoter 4330..4358
/label="AmpR_promoter"
gene 4400..5260
/label="Ampicillin"
/gene="Ampicillin"
CDS 4400..5260
/label="ORF frame 2"
rep_origin 5415..6034
/label="pBR322_origin"
ORIGIN
1 TCAATATTGG CCATTAGCCA TATTATTCAT TGGTTATATA GCATAAATCA ATATTGGCTA
61 TTGGCCATTG CATACGTTGT ATCTATATCA TAATATGTAC ATTTATATTG GCTCATGTCC
121 AATATGACCG CCATGTTGGC ATTGATTATT GACTAGTTAT TAATAGTAAT CAATTACGGG
181 GTCATTAGTT CATAGCCCAT ATATGGAGTT CCGCGTTACA TAACTTACGG TAAATGGCCC
241 GCCTGGCTGA CCGCCCAACG ACCCCCGCCC ATTGACGTCA ATAATGACGT ATGTTCCCAT
301 AGTAACGCCA ATAGGGACTT TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC
361 CCACTTGGCA GTACATCAAG TGTATCATAT GCCAAGTCCG CCCCCTATTG ACGTCAATGA
421 CGGTAAATGG CCCGCCTGGC ATTATGCCCA GTACATGACC TTACGGGACT TTCCTACTTG
481 GCAGTACATC TACGTATTAG TCATCGCTAT TACCATGGTG ATGCGGTTTT GGCAGTACAC
541 CAATGGGCGT GGATAGCGGT TTGACTCACG GGGATTTCCA AGTCTCCACC CCATTGACGT
601 CAATGGGAGT TTGTTTTGGC ACCAAAATCA ACGGGACTTT CCAAAATGTC GTAACAACTG
661 CGATCGCCCG CCCCGTTGAC GCAAATGGGC GGTAGGCGTG TACGGTGGGA GGTCTATATA
721 AGCAGAGCTC GTTTAGTGAA CCGTCAGATC ACTAGAAGCT TTATTGCGGT AGTTTATCAC
781 AGTTAAATTG CTAACGCAGT CAGTGCTTCT GACACAACAG TCTCGAACTT AAGCTGCAGT
841 GACTCTCTTA AGGTAGCCTT GCAGAAGTTG GTCGTGAGGC ACTGGGCAGG TAAGTATCAA
901 GGTTACAAGA CAGGTTTAAG GAGACCAATA GAAACTGGGC TTGTCGAGAC AGAGAAGACT
961 CTTGCGTTTC TGATAGGCAC CTATTGGTCT TACTGACATC CACTTTGCCT TTCTCTCCAC
1021 AGGTGTCCAC TCCCAGTTCA ATTACAGCTC TTAAGGCTAG AGTACTTAAT ACGACTCACT
1081 ATAGGCTAGC CTCGAGAATT CACGCGTCGA GCATGCATCT AGGGCGGCCA ATTCCGCCCC
1141 TCTCCCTCCC CCCCCCCTAA CGTTACTGGC CGAAGCCGCT TGGAATAAGG CCGGTGTGCG
1201 TTTGTCTATA TGTGATTTTC CACCATATTG CCGTCTTTTG GCAATGTGAG GGCCCGGAAA
1261 CCTGGCCCTG TCTTCTTGAC GAGCATTCCT AGGGGTCTTT CCCCTCTCGC CAAAGGAATG
1321 CAAGGTCTGT TGAATGTCGT GAAGGAAGCA GTTCCTCTGG AAGCTTCTTG AAGACAAACA
1381 ACGTCTGTAG CGACCCTTTG CAGGCAGCGG AACCCCCCAC CTGGCGACAG GTGCCTCTGC
1441 GGCCAAAAGC CACGTGTATA AGATACACCT GCAAAGGCGG CACAACCCCA GTGCCACGTT
1501 GTGAGTTGGA TAGTTGTGGA AAGAGTCAAA TGGCTCTCCT CAAGCGTATT CAACAAGGGG
1561 CTGAAGGATG CCCAGAAGGT ACCCCATTGT ATGGGATCTG ATCTGGGGCC TCGGTGCACA
1621 TGCTTTACAT GTGTTTAGTC GAGGTTAAAA AAACGTCTAG GCCCCCCGAA CCACGGGGAC
1681 GTGGTTTTCC TTTGAAAAAC ACGATGATAA GCTTGCCACA ACCCGGGATC CTCTAGAGTC
1741 GACCCGGGCG GCCGCTTCCC TTTAGTGAGG GTTAATGCTT CGAGCAGACA TGATAAGATA
1801 CATTGATGAG TTTGGACAAA CCACAACTAG AATGCAGTGA AAAAAATGCT TTATTTGTGA
1861 AATTTGTGAT GCTATTGCTT TATTTGTAAC CATTATAAGC TGCAATAAAC AAGTTAACAA
1921 CAACAATTGC ATTCATTTTA TGTTTCAGGT TCAGGGGGAG ATGTGGGAGG TTTTTTAAAG
1981 CAAGTAAAAC CTCTACAAAT GTGGTAAAAT CCGATAAGGA TCGATCCGGG CTGGCGTAAT
2041 AGCGAAGAGG CCCGCACCGA TCGCCCTTCC CAACAGTTGC GCAGCCTGAA TGGCGAATGG
2101 ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG CGGGTGTGGT GGTTACGCGC AGCGTGACCG
2161 CTACACTTGC CAGCGCCCTA GCGCCCGCTC CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA
2221 CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA ATCGGGGGCT CCCTTTAGGG TTCCGATTTA
2281 GAGCTTTACG GCACCTCGAC CGCAAAAAAC TTGATTTGGG TGATGGTTCA CGTAGTGGGC
2341 CATCGCCCTG ATAGACGGTT TTTCGCCCTT TGACGTTGGA GTCCACGTTC TTTAATAGTG
2401 GACTCTTGTT CCAAACTGGA ACAACACTCA ACCCTATCTC GGTCTATTCT TTTGATTTAT
2461 AAGGGATTTT GCCGATTTCG GCCTATTGGT TAAAAAATGA GCTGATTTAA CAAATATTTA
2521 ACGCGAATTT TAACAAAATA TTAACGTTTA CAATTTCGCC TGATGCGGTA TTTTCTCCTT
2581 ACGCATCTGT GCGGTATTTC ACACCGCATA CGCGGATCTG CGCAGCACCA TGGCCTGAAA
2641 TAACCTCTGA AAGAGGAACT TGGTTAGGTA CCTTCTGAGG CGGAAAGAAC CAGCTGTGGA
2701 ATGTGTGTCA GTTAGGGTGT GGAAAGTCCC CAGGCTCCCC AGCAGGCAGA AGTATGCAAA
2761 GCATGCATCT CAATTAGTCA GCAACCAGGT GTGGAAAGTC CCCAGGCTCC CCAGCAGGCA
2821 GAAGTATGCA AAGCATGCAT CTCAATTAGT CAGCAACCAT AGTCCCGCCC CTAACTCCGC
2881 CCATCCCGCC CCTAACTCCG CCCAGTTCCG CCCATTCTCC GCCCCATGGC TGACTAATTT
2941 TTTTTATTTA TGCAGAGGCC GAGGCCGCCT CGGCCTCTGA GCTATTCCAG AAGTAGTGAG
3001 GAGGCTTTTT TGGAGGCCTA GGCTTTTGCA AAAAGCTTGA TTCTTCTGAC ACAACAGTCT
3061 CGAACTTAAG GCTAGAGCCA CCATGATTGA ACAAGATGGA TTGCACGCAG GTTCTCCGGC
3121 CGCTTGGGTG GAGAGGCTAT TCGGCTATGA CTGGGCACAA CAGACAATCG GCTGCTCTGA
3181 TGCCGCCGTG TTCCGGCTGT CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA AGACCGACCT
3241 GTCCGGTGCC CTGAATGAAC TGCAGGACGA GGCAGCGCGG CTATCGTGGC TGGCCACGAC
3301 GGGCGTTCCT TGCGCAGCTG TGCTCGACGT TGTCACTGAA GCGGGAAGGG ACTGGCTGCT
3361 ATTGGGCGAA GTGCCGGGGC AGGATCTCCT GTCATCTCAC CTTGCTCCTG CCGAGAAAGT
3421 ATCCATCATG GCTGATGCAA TGCGGCGGCT GCATACGCTT GATCCGGCTA CCTGCCCATT
3481 CGACCACCAA GCGAAACATC GCATCGAGCG AGCACGTACT CGGATGGAAG CCGGTCTTGT
3541 CGATCAGGAT GATCTGGACG AAGAGCATCA GGGGCTCGCG CCAGCCGAAC TGTTCGCCAG
3601 GCTCAAGGCG CGCATGCCCG ACGGCGAGGA TCTCGTCGTG ACCCATGGCG ATGCCTGCTT
3661 GCCGAATATC ATGGTGGAAA ATGGCCGCTT TTCTGGATTC ATCGACTGTG GCCGGCTGGG
3721 TGTGGCGGAC CGCTATCAGG ACATAGCGTT GGCTACCCGT GATATTGCTG AAGAGCTTGG
3781 CGGCGAATGG GCTGACCGCT TCCTCGTGCT TTACGGTATC GCCGCTCCCG ATTCGCAGCG
3841 CATCGCCTTC TATCGCCTTC TTGACGAGTT CTTCTGAGCG GGACTCTGGG GTTCGAAATG
3901 ACCGACCAAG CGACGCCCAA CCTGCCATCA CGATGGCCGC AATAAAATAT CTTTATTTTC
3961 ATTACATCTG TGTGTTGGTT TTTTGTGTGA ATCGATAGCG ATAAGGATCC GCGTATGGTG
4021 CACTCTCAGT ACAATCTGCT CTGATGCCGC ATAGTTAAGC CAGCCCCGAC ACCCGCCAAC
4081 ACCCGCTGAC GCGCCCTGAC GGGCTTGTCT GCTCCCGGCA TCCGCTTACA GACAAGCTGT
4141 GACCGTCTCC GGGAGCTGCA TGTGTCAGAG GTTTTCACCG TCATCACCGA AACGCGCGAG
4201 ACGAAAGGGC CTCGTGATAC GCCTATTTTT ATAGGTTAAT GTCATGATAA TAATGGTTTC
4261 TTAGACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT
4321 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA
4381 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT
4441 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC
4501 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
4561 CCTTGAGAGT TTTCGCCCCG AAGAACGTTT TCCAATGATG AGCACTTTTA AAGTTCTGCT
4621 ATGTGGCGCG GTATTATCCC GTATTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA
4681 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG
4741 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA
4801 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
4861 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA
4921 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG
4981 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT
5041 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG
5101 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
5161 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA
5221 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC
5281 ATATATACTT TAGATTGATT TAAAACTTCA TTTTTAATTT AAAAGGATCT AGGTGAAGAT
5341 CCTTTTTGAT AATCTCATGA CCAAAATCCC TTAACGTGAG TTTTCGTTCC ACTGAGCGTC
5401 AGACCCCGTA GAAAAGATCA AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG
5461 CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT
5521 ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTCCT
5581 TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT
5641 CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG
5701 GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC
5761 GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA
5821 GCTATGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG
5881 CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA
5941 TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG
6001 GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG
6061 CTGGCCTTTT GCTCACATGG CTCGACAGAT CT
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