| 出品公司: | Invitrogen |
|---|---|
| 载体名称: | pIB/V5-His-TOPO |
| 质粒类型: | 昆虫表达载体；TOPO-TA克隆载体 |
| 克隆方法: | TOPO-TA |
| 启动子: | OplE2 |
| 载体大小: | 3537bp |
| 5' 测序引物及序列: | OplE2 Forward：5'd[CGCAACGATCTGGTAAACAC]3' |
| 3' 测序引物及序列: | -- |
| 载体标签: | V5 Epitope Tag, His Tag (C-末端) |
| 载体抗性: | 氨苄青霉素 |
| 真核筛选标记: | Blasticidin |
| 克隆菌株: | TOP10 |
| 宿主细胞（系）: | Sf9，Sf21，HighFive |
| 备注: | -- |
| 产品目录号: | K89020 |
| 瞬表达/稳表达: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
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
描述
The pIB/V5-His TOPO® TA Expression Kit offers five-minute cloning of Taq-amplified PCR products directly into the pIB/V5-His-TOPO expression vector. In addition to being TOPO Cloning ready, pIB/V5-His-TOPO includes:
• The OpIE2 promoter for constitutive expression
• The Blasticidin resistance gene for rapid selection of stably transfected cell lines in two weeks
• C-terminal V5 epitope and polyhistidine (6xHis) sequence for detection with Invitrogen's Anti-V5 Antibody and rapid purification using nickel-chelating resin
表达水平
The OpIE2 promoter provides relatively high levels of constitutive expression, although not all proteins will express as high as might be expected from baculovirus late promoters such as polyhedrin or very late promoters such as p10 (Jarvis et al., 1996).
However, some researchers have found that the InsectSelect™ System expresses some proteins better than baculovirus systems. To date, reported expression levels range from 1-2 µg/ml (human IL-6; Invitrogen) to 8-10 µg/ml (human melanotransferrin) (Hegeduset al., 1999).
载体序列
LOCUS pIB/V5-His-TOPO 3537 bp DNA SYN
DEFINITION pIB/V5-His-TOPO
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..3537
/organism="pIB/V5-His-TOPO"
/mol_type="other DNA"
misc_feature 511..530
/label="OpIE2_fwd_primer"
rep_origin complement(1003..1622)
/label="pBR322_origin"
promoter 1996..2063
/label="EM7_promoter"
gene 2064..2462
/label="blast (variant)"
/gene="blast (variant)"
gene 2582..3442
/label="Ampicillin"
/gene="Ampicillin"
CDS 2582..3442
/label="ORF frame 2"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
ORIGIN
1 CATGATGATA AACAATGTAT GGTGCTAATG TTGCTTCAAC AACAATTCTG TTGAACTGTG
61 TTTTCATGTT TGCCAACAAG CACCTTTATA CTCGGTGGCC TCCCCACCAC CAACTTTTTT
121 GCACTGCAAA AAAACACGCT TTTGCACGCG GGCCCATACA TAGTACAAAC TCTACGTTTC
181 GTAGACTATT TTACATAAAT AGTCTACACC GTTGTATACG CTCCAAATAC ACTACCACAC
241 ATTGAACCTT TTTGCAGTGC AAAAAAGTAC GTGTCGGCAG TCACGTAGGC CGGCCTTATC
301 GGGTCGCGTC CTGTCACGTA CGAATCACAT TATCGGACCG GACGAGTGTT GTCTTATCGT
361 GACAGGACGC CAGCTTCCTG TGTTGCTAAC CGCAGCCGGA CGCAACTCCT TATCGGAACA
421 GGACGCGCCT CCATATCAGC CGCGCGTTAT CTCATGCGCG TGACCGGACA CGAGGCGCCC
481 GTCCCGCTTA TCGCGCCTAT AAATACAGCC CGCAACGATC TGGTAAACAC AGTTGAACAG
541 CATCTGTTCG AATTTAAAGC TTGGTACCGA GCTCGGATCC ACTAGTCCAG TGTGGTGGAA
601 TTGCCCTTAA GGGCAATTCT GCAGATATCC AGCACAGTGG CGGCCGCTCG AGTCTAGAGG
661 GCCCGCGGTT CGAAGGTAAG CCTATCCCTA ACCCTCTCCT CGGTCTCGAT TCTACGCGTA
721 CCGGTCATCA TCACCATCAC CATTGAGTTT ATCTGACTAA ATCTTAGTTT GTATTGTCAT
781 GTTTTAATAC AATATGTTAT GTTTAAATAT GTTTTTAATA AATTTTATAA AATAATTTCA
841 ACTTTTATTG TAACAACATT GTCCATTTAC ACACTCCTTT CAAGCGCGTG GGATCGATGC
901 TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT
961 GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT
1021 CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG
1081 AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC
1141 TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT
1201 GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA
1261 GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA
1321 TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA
1381 CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA
1441 CTACGGCTAC ACTAGAAGAA CAGTATTTGG TATCTGCGCT CTGCTGAAGC CAGTTACCTT
1501 CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT
1561 TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT
1621 CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT
1681 GCGAAACACG CACGGCGCGC GCACGCAGCT TAGCACAAAC GCGTCGTTGC ACGCGCCCAC
1741 CGCTAACCGC AGGCCAATCG GTCGGCCGGC CTCATATCCG CTCACCAGCC GCGTCCTATC
1801 GGGCGCGGCT TCCGCGCCCA TTTTGAATAA ATAAACGATA ACGCCGTTGG TGGCGTGAGG
1861 CATGTAAAAG GTTACATCAT TATCTTGTTC GCCATCCGGT TGGTATAAAT AGACGTTCAT
1921 GTTGGTTTTT GTTTCAGTTG CAAGTTGGCT GCGGCGCGCG CAGCACCTTT GCCGGGATCT
1981 GCCGGGCTGC AGCACGTGTT GACAATTAAT CATCGGCATA GTATATCGGC ATAGTATAAT
2041 ACGACAAGGT GAGGAACTAA ACCATGGCCA AGCCTTTGTC TCAAGAAGAA TCCACCCTCA
2101 TTGAAAGAGC AACGGCTACA ATCAACAGCA TCCCCATCTC TGAAGACTAC AGCGTCGCCA
2161 GCGCAGCTCT CTCTAGCGAC GGCCGCATCT TCACTGGTGT CAATGTATAT CATTTTACTG
2221 GGGGACCTTG CGCAGAACTC GTGGTGCTGG GCACTGCTGC TGCTGCGGCA GCTGGCAACC
2281 TGACTTGTAT CGTCGCGATC GGAAATGAGA ACAGGGGCAT CTTGAGCCCC TGCGGACGGT
2341 GCCGACAGGT TCTTCTCGAT CTGCATCCTG GGATCAAAGC CATAGTGAAG GACAGTGATG
2401 GACAGCCGAC GGCAGTTGGG ATTCGTGAAT TGCTGCCCTC TGGTTATGTG TGGGAGGGCT
2461 AAGCACTTCG TGGCCGAGGA GCAGGACTGA CACGTCCCGG GAGATCTGCA TGTCTACTAA
2521 ACTCACAAAT TAGAGCTTCA ATTTAATTAT ATCAGTTATT ACCCATTGAA AAAGGAAGAG
2581 TATGAGTATT CAACATTTCC GTGTCGCCCT TATTCCCTTT TTTGCGGCAT TTTGCCTTCC
2641 TGTTTTTGCT CACCCAGAAA CGCTGGTGAA AGTAAAAGAT GCTGAAGATC AGTTGGGTGC
2701 ACGAGTGGGT TACATCGAAC TGGATCTCAA CAGCGGTAAG ATCCTTGAGA GTTTTCGCCC
2761 CGAAGAACGT TTTCCAATGA TGAGCACTTT TAAAGTTCTG CTATGTGGCG CGGTATTATC
2821 CCGTATTGAC GCCGGGCAAG AGCAACTCGG TCGCCGCATA CACTATTCTC AGAATGACTT
2881 GGTTGAGTAC TCACCAGTCA CAGAAAAGCA TCTTACGGAT GGCATGACAG TAAGAGAATT
2941 ATGCAGTGCT GCCATAACCA TGAGTGATAA CACTGCGGCC AACTTACTTC TGACAACGAT
3001 CGGAGGACCG AAGGAGCTAA CCGCTTTTTT GCACAACATG GGGGATCATG TAACTCGCCT
3061 TGATCGTTGG GAACCGGAGC TGAATGAAGC CATACCAAAC GACGAGCGTG ACACCACGAT
3121 GCCTGTAGCA ATGGCAACAA CGTTGCGCAA ACTATTAACT GGCGAACTAC TTACTCTAGC
3181 TTCCCGGCAA CAATTAATAG ACTGGATGGA GGCGGATAAA GTTGCAGGAC CACTTCTGCG
3241 CTCGGCCCTT CCGGCTGGCT GGTTTATTGC TGATAAATCT GGAGCCGGTG AGCGTGGGTC
3301 TCGCGGTATC ATTGCAGCAC TGGGGCCAGA TGGTAAGCCC TCCCGTATCG TAGTTATCTA
3361 CACGACGGGG AGTCAGGCAA CTATGGATGA ACGAAATAGA CAGATCGCTG AGATAGGTGC
3421 CTCACTGATT AAGCATTGGT AACTGTCAGA CCAAGTTTAC TCATATATAC TTTAGATTGA
3481 TTTAAAACTT CATTTTTAAT TTAAAAGGAT CTAGGTGAAG ATCCTTTTTG ATAATCT
//
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