出品公司: | Invitrogen |
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載體名稱: | pBAD/Myc-His C |
質粒類型: | 大腸桿菌表達載體;誘導表達載體 |
高拷貝/低拷貝: | 低拷貝 |
克隆方法: | 限制性內切酶;多克隆位點 |
啟動子: | araBAD |
載體大小: | 4093 bp |
5' 測序引物及序列: | pBAD Forward: 5′-ATGCCATAGCATTTTTATCC-3′ |
3' 測序引物及序列: | pBAD Reverse 5′-GATTTAATCTGTATCAGG-3′ |
載體標簽: | 6x His Tag(C-端),c-Myc Epitope(C-端) |
載體抗性: | 氨芐青霉素(Ampicillin) |
克隆菌株: | TOP10 |
表達菌株: | 推薦LMG194 |
備注: |
pBAD/Myc-His C載體是阿拉伯糖調控載體; 在無葡萄糖的培養基中,阿拉伯糖正向調控目的基因的表達; 通過調節阿拉伯糖的濃度水平來優化目的蛋白的可溶性表達。 |
產品目錄號: | V440-01 |
穩定性: | 穩表達 |
組成型/誘導型: | 誘導型(阿拉伯糖) |
病毒/非病毒: | 非病毒 |
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pBad/Myc-His C載體 pBadMyc-His C質粒基本信息
pBad/Myc-His C質粒圖譜 pBadMyc-His C載體圖譜和pBad/Myc-His C載體序列 pBadMyc-His C質粒序列多克隆位點信息
pBad/Myc-His C質粒 pBadMyc-His C載體簡介
pBAD/His和PBAD/Myc-His載體質粒是衍生于pBR322載體。載體設計用來在大腸桿菌中進行可調節,劑量依賴性的表達和純化重組目的蛋白。使用大腸桿菌araBAD啟動子(pBAD)增強了大腸桿菌重組蛋白可溶性表達的水平。pBAD/His和pBAD/Myc His載體上的調節蛋白AraC能夠調控pBad啟動子。
pBAD/Myc-His A,B,C 載體簡介
The pBAD/His and pBAD/Myc-His plasmids are pBR322-derived expression vectors designed for regulated, dose-dependent recombinant protein expression and purification in E. coli. Optimum levels of soluble, recombinant protein are possible using the araBAD promoter (PBAD) from E. coli. The regulatory protein, AraC, is provided on the pBAD/His and pBAD/Myc-His vectors allowing regulation of PBAD.
The pBAD/Myc-His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion. The pBAD/Myc-His vector expresses native proteins or fusion proteins with a C-terminal tag. The vector provides:
The araBAD promoter for tightly regulated expression
Translation initiation signals optimized for E. coliexpression
C-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin or detection with an Anti-His(C-term) Antibody
C-terminal c-myc epitope for detection and analysis with an Anti-myc Antibody
Three vectors are provided (A, B, and C). Each has the C-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.
L-阿拉伯糖調控表達
In the presence of L-arabinose, expression from PBAD is turned on while the absence of L-arabinose produces very low levels of transcription from PBAD (Lee, 1980; Lee et al., 1987). Uninduced levels are repressed even further by growth in the presence of glucose. Glucose reduces the levels of 3′,5′-cyclic AMP, thus lowering expression of the catabolite-repressed PBAD promoter (Miyada et al., 1984). By varying the concentration of L-arabinose, protein expression levels can be optimized to ensure maximum expression of soluble protein. In addition, the tight regulation of PBAD by AraC is useful for expression of potentially toxic or essential genes (Carson et al., 1991; Dalbey and Wickner, 1985; Guzman et al., 1992; Kuhn and Wickner, 1985; Russell et al., 1989; San Millan et al., 1989). For more information on the mechanism of expression and repression of the ara regulon, refer to Schleif, 1992.
pBad/Myc-His C質粒序列 pBadMyc-His C載體序列
LOCUS pBAD/myc-His C 4093 bp DNA circular SYN DEFINITION pBAD/myc-His C ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. COMMENT This file is created by Vector NTI //gt12365.cn/ COMMENT VNTAUTHORNAME|gt12365.cn| FEATURES Location/Qualifiers source 1..4093 /organism="pBAD/myc-His C" /mol_type="other DNA" misc_feature 4..19 /label="araO2" misc_feature 161..172 /label="araO1" misc_feature 203..216 /label="CAP_BS" misc_feature 208..227 /label="pBAD_fwd_primer" misc_feature 213..251 /label="AraI1I2" promoter 248..276 /label="ARA_promoter" misc_feature complement(494..511) /label="pTrcHis_rev_primer" misc_feature complement(494..511) /label="pBAD_rev_primer" terminator 544..701 /label="rrnB_terminator" terminator 667..710 /label="rrnB_T1_terminator" terminator 842..869 /label="rrnB_T2_terminator" promoter 910..938 /label="AmpR_promoter" gene 980..1840 /label="Ampicillin" /gene="Ampicillin" CDS 980..1840 /label="ORF frame 2" rep_origin 1995..2614 /label="pBR322_origin" misc_feature 3011..3033 /label="pGEX_3_primer" misc_feature complement(3189..4067) /label="AraC" CDS complement(3189..25) /label="ORF frame 1" CDS complement(3189..4076) /label="ORF frame 3" ORIGIN 1 AAGAAACCAA TTGTCCATAT TGCATCAGAC ATTGCCGTCA CTGCGTCTTT TACTGGCTCT 61 TCTCGCTAAC CAAACCGGTA ACCCCGCTTA TTAAAAGCAT TCTGTAACAA AGCGGGACCA 121 AAGCCATGAC AAAAACGCGT AACAAAAGTG TCTATAATCA CGGCAGAAAA GTCCACATTG 181 ATTATTTGCA CGGCGTCACA CTTTGCTATG CCATAGCATT TTTATCCATA AGATTAGCGG 241 ATCCTACCTG ACGCTTTTTA TCGCAACTCT CTACTGTTTC TCCATACCCG TTTTTTGGGC 301 TAACAGGAGG AATTAACCAT GGATCCGAGC TCGAGATCTG CAGCTGGTAC CATATGGGAA 361 TTCGAAGCTT ACGTAGAACA AAAACTCATC TCAGAAGAGG ATCTGAATAG CGCCGTCGAC 421 CATCATCATC ATCATCATTG AGTTTAAACG GTCTCCAGCT TGGCTGTTTT GGCGGATGAG 481 AGAAGATTTT CAGCCTGATA CAGATTAAAT CAGAACGCAG AAGCGGTCTG ATAAAACAGA 541 ATTTGCCTGG CGGCAGTAGC GCGGTGGTCC CACCTGACCC CATGCCGAAC TCAGAAGTGA 601 AACGCCGTAG CGCCGATGGT AGTGTGGGGT CTCCCCATGC GAGAGTAGGG AACTGCCAGG 661 CATCAAATAA AACGAAAGGC TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG 721 TCGGTGAACG CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG 781 CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGCA TCAAATTAAG 841 CAGAAGGCCA TCCTGACGGA TGGCCTTTTT GCGTTTCTAC AAACTCTTTT GTTTATTTTT 901 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA 961 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT 1021 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC 1081 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT 1141 CCTTGAGAGT TTTCGCCCCG AAGAACGTTT TCCAATGATG AGCACTTTTA AAGTTCTGCT 1201 ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA 1261 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG 1321 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA 1381 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG 1441 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA 1501 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG 1561 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT 1621 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG 1681 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC 1741 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA 1801 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC 1861 ATATATACTT TAGATTGATT TAAAACTTCA TTTTTAATTT AAAAGGATCT AGGTGAAGAT 1921 CCTTTTTGAT AATCTCATGA CCAAAATCCC TTAACGTGAG TTTTCGTTCC ACTGAGCGTC 1981 AGACCCCGTA GAAAAGATCA AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG 2041 CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT 2101 ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTCCT 2161 TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT 2221 CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG 2281 GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC 2341 GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA 2401 GCTATGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG 2461 CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA 2521 TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG 2581 GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG 2641 CTGGCCTTTT GCTCACATGT TCTTTCCTGC GTTATCCCCT GATTCTGTGG ATAACCGTAT 2701 TACCGCCTTT GAGTGAGCTG ATACCGCTCG CCGCAGCCGA ACGACCGAGC GCAGCGAGTC 2761 AGTGAGCGAG GAAGCGGAAG AGCGCCTGAT GCGGTATTTT CTCCTTACGC ATCTGTGCGG 2821 TATTTCACAC CGCATATGGT GCACTCTCAG TACAATCTGC TCTGATGCCG CATAGTTAAG 2881 CCAGTATACA CTCCGCTATC GCTACGTGAC TGGGTCATGG CTGCGCCCCG ACACCCGCCA 2941 ACACCCGCTG ACGCGCCCTG ACGGGCTTGT CTGCTCCCGG CATCCGCTTA CAGACAAGCT 3001 GTGACCGTCT CCGGGAGCTG CATGTGTCAG AGGTTTTCAC CGTCATCACC GAAACGCGCG 3061 AGGCAGCAGA TCAATTCGCG CGCGAAGGCG AAGCGGCATG CATAATGTGC CTGTCAAATG 3121 GACGAAGCAG GGATTCTGCA AACCCTATGC TACTCCGTCA AGCCGTCAAT TGTCTGATTC 3181 GTTACCAATT ATGACAACTT GACGGCTACA TCATTCACTT TTTCTTCACA ACCGGCACGG 3241 AACTCGCTCG GGCTGGCCCC GGTGCATTTT TTAAATACCC GCGAGAAATA GAGTTGATCG 3301 TCAAAACCAA CATTGCGACC GACGGTGGCG ATAGGCATCC GGGTGGTGCT CAAAAGCAGC 3361 TTCGCCTGGC TGATACGTTG GTCCTCGCGC CAGCTTAAGA CGCTAATCCC TAACTGCTGG 3421 CGGAAAAGAT GTGACAGACG CGACGGCGAC AAGCAAACAT GCTGTGCGAC GCTGGCGATA 3481 TCAAAATTGC TGTCTGCCAG GTGATCGCTG ATGTACTGAC AAGCCTCGCG TACCCGATTA 3541 TCCATCGGTG GATGGAGCGA CTCGTTAATC GCTTCCATGC GCCGCAGTAA CAATTGCTCA 3601 AGCAGATTTA TCGCCAGCAG CTCCGAATAG CGCCCTTCCC CTTGCCCGGC GTTAATGATT 3661 TGCCCAAACA GGTCGCTGAA ATGCGGCTGG TGCGCTTCAT CCGGGCGAAA GAACCCCGTA 3721 TTGGCAAATA TTGACGGCCA GTTAAGCCAT TCATGCCAGT AGGCGCGCGG ACGAAAGTAA 3781 ACCCACTGGT GATACCATTC GCGAGCCTCC GGATGACGAC CGTAGTGATG AATCTCTCCT 3841 GGCGGGAACA GCAAAATATC ACCCGGTCGG CAAACAAATT CTCGTCCCTG ATTTTTCACC 3901 ACCCCCTGAC CGCGAATGGT GAGATTGAGA ATATAACCTT TCATTCCCAG CGGTCGGTCG 3961 ATAAAAAAAT CGAGATAACC GTTGGCCTCA ATCGGCGTTA AACCCGCCAC CAGATGGGCA 4021 TTAAACGAGT ATCCCGGCAG CAGGGGATCA TTTTGCGCTT CAGCCATACT TTTCATACTC 4081 CCGCCATTCA GAG //
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