| 出品公司: | |
|---|---|
| 载体名称: | pBAD/His A |
| 质粒类型: | 大肠杆菌表达载体；诱导表达载体 |
| 高拷贝/低拷贝: | 低拷贝 |
| 克隆方法: | 限制性内切酶；多克隆位点 |
| 启动子: | araBAD |
| 载体大小: | 4102 bp |
| 5' 测序引物及序列: | pBAD Forward: 5′-ATGCCATAGCATTTTTATCC-3′ |
| 3' 测序引物及序列: | pBAD Reverse 5′-GATTTAATCTGTATCAGG-3′ |
| 载体标签: | 6x His Tag（N-端）,Xpress Epitope Tag（N-端）, EK 切割位点 |
| 载体抗性: | 氨苄青霉素（Ampicillin） |
| 克隆菌株: | TOP10 |
| 表达菌株: | 推荐LMG194 |
| 备注: | pBAD/His A载体是阿拉伯糖调控载体； 在无葡萄糖的培养基中，阿拉伯糖正向调控目的基因 的表达； 通过调节阿拉伯糖的浓度水平来优化目的蛋白的可溶性表达； pBAD/His A,B,C之间仅阅读框不同。 |
| 产品目录号: | ZY430-01 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型（阿拉伯糖) |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pBad/His A
- 库存：
20
- 供应商：
泽叶生物
pBadHis A载体基本信息
pBadHis A载体质粒图谱和多克隆位点信息
pBadHis A载体简介
pBAD/His和PBAD/Myc-His载体质粒是衍生于pBR322载体。载体设计用来在大肠杆菌中进行可调节，剂量依赖性的表达和纯化重组目的蛋白。使用大肠杆菌araBAD启动子（pBAD）增强了大肠杆菌重组蛋白可溶性表达的水平。pBAD/His和pBAD/Myc His载体上的调节蛋白AraC能够调控pBad启动子。
pBAD/His A,B,C 载体介绍
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-阿拉伯糖调控表达
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.
pBadHis A载体序列
LOCUS pBAD/His A 4102 bp DNA circular SYN
DEFINITION pBAD/His A
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..4102
/organism="pBAD/His A"
/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 351..369
/label="Xpress_fwd_primer"
misc_feature 352..384
/label="T7_leader"
misc_feature 388..411
/label="Xpress_EK"
misc_feature complement(503..520)
/label="pBAD_rev_primer"
misc_feature complement(503..520)
/label="pTrcHis_rev_primer"
terminator 553..710
/label="rrnB_terminator"
terminator 676..719
/label="rrnB_T1_terminator"
terminator 851..878
/label="rrnB_T2_terminator"
promoter 919..947
/label="AmpR_promoter"
gene 989..1849
/label="Ampicillin"
/gene="Ampicillin"
CDS 989..1849
/label="ORF frame 2"
rep_origin 2004..2623
/label="pBR322_origin"
misc_feature 3020..3042
/label="pGEX_3_primer"
misc_feature complement(3198..4076)
/label="AraC"
CDS complement(3198..25)
/label="ORF frame 1"
CDS complement(3198..4085)
/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 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
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pBad/His A载体
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