pIB/V5-His-TOPO

pIB/V5-His-TOPO

载体基本信息

出品公司:	Ybscience
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:	--
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

 

载体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