請(qǐng)?jiān)诰€咨詢或直接聯(lián)系18171096062/18986294316
SMART 快構(gòu) 120元-請(qǐng)注冊(cè)登錄 http://49.234.53.21/login
基本信息
載體類型: | 大腸桿菌表達(dá)載體;誘導(dǎo)表達(dá)載體 |
載體抗性: | Amp |
載體標(biāo)簽: | N-端:6xHis, Xpress Epitope, EK切割位點(diǎn) |
啟動(dòng)子: | araBAD |
高拷貝/低拷貝: | 低拷貝 |
載體大小: | 4857 bp |
克隆方法: | 多克隆位點(diǎn),限制性內(nèi)切酶 |
5' 測(cè)序引物: | pBAD Forward: 5′-ATGCCATAGCATTTTTATCC-3′ |
3' 測(cè)序引物: | pBAD Reverse 5′-GATTTAATCTGTATCAGG-3′ |
備注: | 如有侵權(quán)請(qǐng)產(chǎn)權(quán)持有人盡快聯(lián)系法務(wù)15927229196 |
質(zhì)粒圖譜和多克隆位點(diǎn)信息
注意:
客戶在我公司選擇構(gòu)建載體服務(wù)項(xiàng)目時(shí),請(qǐng)自行提供母源載體。
如要使用本公司保存的母源載體時(shí),本公司不能保證所有母源載體完全符合客戶預(yù)期,也不承擔(dān)由此產(chǎn)生的任何責(zé)任。
載體描述
pBAD/His和PBAD/Myc-His載體質(zhì)粒是衍生于pBR322載體。載體設(shè)計(jì)用來(lái)在大腸桿菌中進(jìn)行可調(diào)節(jié),劑量依賴性的表達(dá)和純化重組目的蛋白。使用大腸桿菌araBAD啟動(dòng)子(pBAD)增強(qiáng)了大腸桿菌重組蛋白可溶性表達(dá)的水平。pBAD/His和pBAD/Myc His載體上的調(diào)節(jié)蛋白AraC能夠調(diào)控pBad啟動(dòng)子。
pBAD/His A,B,C 載體介紹
The pBAD/His Kit provides all of the necessary reagents to express your protein in a tightly regulated fashion.
The vector pBAD/His allows you to express your protein with an N-terminal tag. The vector provides:
The araBAD promoter for tightly regulated expression
Translation initiation signals optimized for E. coliexpression
N-terminal polyhistidine (6xHis) tag for purification with nickel-chelating resin and detection with an Anti-
HisG Antibody
N-terminal Xpress epitope for detection and analysis with an Anti-Xpress Antibody
Enterokinase cleavage site for removing the N-terminal tag following purification
Three vectors are provided (A, B, and C). Each has the N-terminal tag in a different reading frame relative to the multiple cloning site to simplify in-frame cloning of your gene.
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.
L-阿拉伯糖調(diào)控表達(dá)
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.
載體序列
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 GGGGGGTTCT CATCATCATC ATCATCATGG TATGGCTAGC
361 ATGACTGGTG GACAGCAAAT GGGTCGGGAT CTGTACGACG ATGACGATAA GGATCGATGG
421 GGATCCGAGC TCGAGATCTG CAGCTGGTAC CATATGGGAA TTCGAAGCTT GGCTGTTTTG
481 GCGGATGAGA GAAGATTTTC AGCCTGATAC AGATTAAATC AGAACGCAGA AGCGGTCTGA
541 TAAAACAGAA TTTGCCTGGC GGCAGTAGCG CGGTGGTCCC ACCTGACCCC ATGCCGAACT
601 CAGAAGTGAA ACGCCGTAGC GCCGATGGTA GTGTGGGGTC TCCCCATGCG AGAGTAGGGA
661 ACTGCCAGGC ATCAAATAAA ACGAAAGGCT CAGTCGAAAG ACTGGGCCTT TCGTTTTATC
721 TGTTGTTTGT CGGTGAACGC TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC
781 GTTGCGAAGC AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGCAT
841 CAAATTAAGC AGAAGGCCAT CCTGACGGAT GGCCTTTTTG CGTTTCTACA AACTCTTTTG
901 TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT
961 GCTTCAATAA TATTGAAAAA GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT
1021 TCCCTTTTTT GCGGCATTTT GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT
1081 AAAAGATGCT GAAGATCAGT TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG
1141 CGGTAAGATC CTTGAGAGTT TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA
1201 AGTTCTGCTA TGTGGCGCGG TATTATCCCG TGTTGACGCC GGGCAAGAGC AACTCGGTCG
1261 CCGCATACAC TATTCTCAGA ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT
1321 TACGGATGGC ATGACAGTAA GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC
1381 TGCGGCCAAC TTACTTCTGA CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA
1441 CAACATGGGG GATCATGTAA CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT
1501 ACCAAACGAC GAGCGTGACA CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT
1561 ATTAACTGGC GAACTACTTA CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC
1621 GGATAAAGTT GCAGGACCAC TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA
1681 TAAATCTGGA GCCGGTGAGC GTGGGTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG
1741 TAAGCCCTCC CGTATCGTAG TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG
1801 AAATAGACAG ATCGCTGAGA TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA
1861 AGTTTACTCA TATATACTTT AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA
1921 GGTGAAGATC CTTTTTGATA ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA
1981 CTGAGCGTCA GACCCCGTAG AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG
2041 CGTAATCTGC TGCTTGCAAA CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA
2101 TCAAGAGCTA CCAACTCTTT TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA
2161 TACTGTCCTT CTAGTGTAGC CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC
2221 TACATACCTC GCTCTGCTAA TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG
2281 TCTTACCGGG TTGGACTCAA GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC
2341 GGGGGGTTCG TGCACACAGC CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT
2401 ACAGCGTGAG CTATGAGAAA GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC
2461 GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG
2521 GTATCTTTAT AGTCCTGTCG GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG
2581 CTCGTCAGGG GGGCGGAGCC TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT
2641 GGCCTTTTGC TGGCCTTTTG CTCACATGTT CTTTCCTGCG TTATCCCCTG ATTCTGTGGA
2701 TAACCGTATT ACCGCCTTTG AGTGAGCTGA TACCGCTCGC CGCAGCCGAA CGACCGAGCG
2761 CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA GCGCCTGATG CGGTATTTTC TCCTTACGCA
2821 TCTGTGCGGT ATTTCACACC GCATATGGTG CACTCTCAGT ACAATCTGCT CTGATGCCGC
2881 ATAGTTAAGC CAGTATACAC TCCGCTATCG CTACGTGACT GGGTCATGGC TGCGCCCCGA
2941 CACCCGCCAA CACCCGCTGA CGCGCCCTGA CGGGCTTGTC TGCTCCCGGC ATCCGCTTAC
3001 AGACAAGCTG TGACCGTCTC CGGGAGCTGC ATGTGTCAGA GGTTTTCACC GTCATCACCG
3061 AAACGCGCGA GGCAGCAGAT CAATTCGCGC GCGAAGGCGA AGCGGCATGC ATAATGTGCC
3121 TGTCAAATGG ACGAAGCAGG GATTCTGCAA ACCCTATGCT ACTCCGTCAA GCCGTCAATT
3181 GTCTGATTCG TTACCAATTA TGACAACTTG ACGGCTACAT CATTCACTTT TTCTTCACAA
3241 CCGGCACGGA ACTCGCTCGG GCTGGCCCCG GTGCATTTTT TAAATACCCG CGAGAAATAG
3301 AGTTGATCGT CAAAACCAAC ATTGCGACCG ACGGTGGCGA TAGGCATCCG GGTGGTGCTC
3361 AAAAGCAGCT TCGCCTGGCT GATACGTTGG TCCTCGCGCC AGCTTAAGAC GCTAATCCCT
3421 AACTGCTGGC GGAAAAGATG TGACAGACGC GACGGCGACA AGCAAACATG CTGTGCGACG
3481 CTGGCGATAT CAAAATTGCT GTCTGCCAGG TGATCGCTGA TGTACTGACA AGCCTCGCGT
3541 ACCCGATTAT CCATCGGTGG ATGGAGCGAC TCGTTAATCG CTTCCATGCG CCGCAGTAAC
3601 AATTGCTCAA GCAGATTTAT CGCCAGCAGC TCCGAATAGC GCCCTTCCCC TTGCCCGGCG
3661 TTAATGATTT GCCCAAACAG GTCGCTGAAA TGCGGCTGGT GCGCTTCATC CGGGCGAAAG
3721 AACCCCGTAT TGGCAAATAT TGACGGCCAG TTAAGCCATT CATGCCAGTA GGCGCGCGGA
3781 CGAAAGTAAA CCCACTGGTG ATACCATTCG CGAGCCTCCG GATGACGACC GTAGTGATGA
3841 ATCTCTCCTG GCGGGAACAG CAAAATATCA CCCGGTCGGC AAACAAATTC TCGTCCCTGA
3901 TTTTTCACCA CCCCCTGACC GCGAATGGTG AGATTGAGAA TATAACCTTT CATTCCCAGC
3961 GGTCGGTCGA TAAAAAAATC GAGATAACCG TTGGCCTCAA TCGGCGTTAA ACCCGCCACC
4021 AGATGGGCAT TAAACGAGTA TCCCGGCAGC AGGGGATCAT TTTGCGCTTC AGCCATACTT
4081 TTCATACTCC CGCCATTCAG AG
//