pBad/His C

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pBadHis C载体基本信息

出品公司: Invitrogen
载体名称: pBAD/His C
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: 4100 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(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 C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: V430-01
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

pBadHis C载体图谱质粒图谱和多克隆位点信息

pBAD-His 载体图谱



pBAD-His C 多克隆位点

pBAD-His 载体特征1
pBAD-His 载体特征2

pBadHis C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.
pBAD-His A,B,C 使用流程



pBadHis C载体序列

LOCUS       pBAD/His C	4100 bp 	DNA	circular	SYN
DEFINITION  pBAD/His C
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..4100
                     /organism="pBAD/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    351..369
                     /label="Xpress_fwd_primer"
     misc_feature    352..384
                     /label="T7_leader"
     misc_feature    388..411
                     /label="Xpress_EK"
     misc_feature    complement(501..518)
                     /label="pBAD_rev_primer"
     misc_feature    complement(501..518)
                     /label="pTrcHis_rev_primer"
     terminator      551..708
                     /label="rrnB_terminator"
     terminator      674..717
                     /label="rrnB_T1_terminator"
     terminator      849..876
                     /label="rrnB_T2_terminator"
     promoter        917..945
                     /label="AmpR_promoter"
     gene            987..1847
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             987..1847
                     /label="ORF frame 3"
                     
     rep_origin      2002..2621
                     /label="pBR322_origin"
                     
     misc_feature    3018..3040
                     /label="pGEX_3_primer"
                     
     misc_feature    complement(3196..4074)
                     /label="AraC"
                     
     CDS             complement(3196..25)
                     /label="ORF frame 2"
                     
     CDS             complement(3196..4083)
                     /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 ATCCGACCTC GAGATCTGCA GATGGTACCA TATGGGAATT CGAAGCTTGG CTGTTTTGGC
  481 GGATGAGAGA AGATTTTCAG CCTGATACAG ATTAAATCAG AACGCAGAAG CGGTCTGATA
  541 AAACAGAATT TGCCTGGCGG CAGTAGCGCG GTGGTCCCAC CTGACCCCAT GCCGAACTCA
  601 GAAGTGAAAC GCCGTAGCGC CGATGGTAGT GTGGGGTCTC CCCATGCGAG AGTAGGGAAC
  661 TGCCAGGCAT CAAATAAAAC GAAAGGCTCA GTCGAAAGAC TGGGCCTTTC GTTTTATCTG
  721 TTGTTTGTCG GTGAACGCTC TCCTGAGTAG GACAAATCCG CCGGGAGCGG ATTTGAACGT
  781 TGCGAAGCAA CGGCCCGGAG GGTGGCGGGC AGGACGCCCG CCATAAACTG CCAGGCATCA
  841 AATTAAGCAG AAGGCCATCC TGACGGATGG CCTTTTTGCG TTTCTACAAA CTCTTTTGTT
  901 TATTTTTCTA AATACATTCA AATATGTATC CGCTCATGAG ACAATAACCC TGATAAATGC
  961 TTCAATAATA TTGAAAAAGG AAGAGTATGA GTATTCAACA TTTCCGTGTC GCCCTTATTC
 1021 CCTTTTTTGC GGCATTTTGC CTTCCTGTTT TTGCTCACCC AGAAACGCTG GTGAAAGTAA
 1081 AAGATGCTGA AGATCAGTTG GGTGCACGAG TGGGTTACAT CGAACTGGAT CTCAACAGCG
 1141 GTAAGATCCT TGAGAGTTTT CGCCCCGAAG AACGTTTTCC AATGATGAGC ACTTTTAAAG
 1201 TTCTGCTATG TGGCGCGGTA TTATCCCGTG TTGACGCCGG GCAAGAGCAA CTCGGTCGCC
 1261 GCATACACTA TTCTCAGAAT GACTTGGTTG AGTACTCACC AGTCACAGAA AAGCATCTTA
 1321 CGGATGGCAT GACAGTAAGA GAATTATGCA GTGCTGCCAT AACCATGAGT GATAACACTG
 1381 CGGCCAACTT ACTTCTGACA ACGATCGGAG GACCGAAGGA GCTAACCGCT TTTTTGCACA
 1441 ACATGGGGGA TCATGTAACT CGCCTTGATC GTTGGGAACC GGAGCTGAAT GAAGCCATAC
 1501 CAAACGACGA GCGTGACACC ACGATGCCTG TAGCAATGGC AACAACGTTG CGCAAACTAT
 1561 TAACTGGCGA ACTACTTACT CTAGCTTCCC GGCAACAATT AATAGACTGG ATGGAGGCGG
 1621 ATAAAGTTGC AGGACCACTT CTGCGCTCGG CCCTTCCGGC TGGCTGGTTT ATTGCTGATA
 1681 AATCTGGAGC CGGTGAGCGT GGGTCTCGCG GTATCATTGC AGCACTGGGG CCAGATGGTA
 1741 AGCCCTCCCG TATCGTAGTT ATCTACACGA CGGGGAGTCA GGCAACTATG GATGAACGAA
 1801 ATAGACAGAT CGCTGAGATA GGTGCCTCAC TGATTAAGCA TTGGTAACTG TCAGACCAAG
 1861 TTTACTCATA TATACTTTAG ATTGATTTAA AACTTCATTT TTAATTTAAA AGGATCTAGG
 1921 TGAAGATCCT TTTTGATAAT CTCATGACCA AAATCCCTTA ACGTGAGTTT TCGTTCCACT
 1981 GAGCGTCAGA CCCCGTAGAA AAGATCAAAG GATCTTCTTG AGATCCTTTT TTTCTGCGCG
 2041 TAATCTGCTG CTTGCAAACA AAAAAACCAC CGCTACCAGC GGTGGTTTGT TTGCCGGATC
 2101 AAGAGCTACC AACTCTTTTT CCGAAGGTAA CTGGCTTCAG CAGAGCGCAG ATACCAAATA
 2161 CTGTCCTTCT AGTGTAGCCG TAGTTAGGCC ACCACTTCAA GAACTCTGTA GCACCGCCTA
 2221 CATACCTCGC TCTGCTAATC CTGTTACCAG TGGCTGCTGC CAGTGGCGAT AAGTCGTGTC
 2281 TTACCGGGTT GGACTCAAGA CGATAGTTAC CGGATAAGGC GCAGCGGTCG GGCTGAACGG
 2341 GGGGTTCGTG CACACAGCCC AGCTTGGAGC GAACGACCTA CACCGAACTG AGATACCTAC
 2401 AGCGTGAGCT ATGAGAAAGC GCCACGCTTC CCGAAGGGAG AAAGGCGGAC AGGTATCCGG
 2461 TAAGCGGCAG GGTCGGAACA GGAGAGCGCA CGAGGGAGCT TCCAGGGGGA AACGCCTGGT
 2521 ATCTTTATAG TCCTGTCGGG TTTCGCCACC TCTGACTTGA GCGTCGATTT TTGTGATGCT
 2581 CGTCAGGGGG GCGGAGCCTA TGGAAAAACG CCAGCAACGC GGCCTTTTTA CGGTTCCTGG
 2641 CCTTTTGCTG GCCTTTTGCT CACATGTTCT TTCCTGCGTT ATCCCCTGAT TCTGTGGATA
 2701 ACCGTATTAC CGCCTTTGAG TGAGCTGATA CCGCTCGCCG CAGCCGAACG ACCGAGCGCA
 2761 GCGAGTCAGT GAGCGAGGAA GCGGAAGAGC GCCTGATGCG GTATTTTCTC CTTACGCATC
 2821 TGTGCGGTAT TTCACACCGC ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT
 2881 AGTTAAGCCA GTATACACTC CGCTATCGCT ACGTGACTGG GTCATGGCTG CGCCCCGACA
 2941 CCCGCCAACA CCCGCTGACG CGCCCTGACG GGCTTGTCTG CTCCCGGCAT CCGCTTACAG
 3001 ACAAGCTGTG ACCGTCTCCG GGAGCTGCAT GTGTCAGAGG TTTTCACCGT CATCACCGAA
 3061 ACGCGCGAGG CAGCAGATCA ATTCGCGCGC GAAGGCGAAG CGGCATGCAT AATGTGCCTG
 3121 TCAAATGGAC GAAGCAGGGA TTCTGCAAAC CCTATGCTAC TCCGTCAAGC CGTCAATTGT
 3181 CTGATTCGTT ACCAATTATG ACAACTTGAC GGCTACATCA TTCACTTTTT CTTCACAACC
 3241 GGCACGGAAC TCGCTCGGGC TGGCCCCGGT GCATTTTTTA AATACCCGCG AGAAATAGAG
 3301 TTGATCGTCA AAACCAACAT TGCGACCGAC GGTGGCGATA GGCATCCGGG TGGTGCTCAA
 3361 AAGCAGCTTC GCCTGGCTGA TACGTTGGTC CTCGCGCCAG CTTAAGACGC TAATCCCTAA
 3421 CTGCTGGCGG AAAAGATGTG ACAGACGCGA CGGCGACAAG CAAACATGCT GTGCGACGCT
 3481 GGCGATATCA AAATTGCTGT CTGCCAGGTG ATCGCTGATG TACTGACAAG CCTCGCGTAC
 3541 CCGATTATCC ATCGGTGGAT GGAGCGACTC GTTAATCGCT TCCATGCGCC GCAGTAACAA
 3601 TTGCTCAAGC AGATTTATCG CCAGCAGCTC CGAATAGCGC CCTTCCCCTT GCCCGGCGTT
 3661 AATGATTTGC CCAAACAGGT CGCTGAAATG CGGCTGGTGC GCTTCATCCG GGCGAAAGAA
 3721 CCCCGTATTG GCAAATATTG ACGGCCAGTT AAGCCATTCA TGCCAGTAGG CGCGCGGACG
 3781 AAAGTAAACC CACTGGTGAT ACCATTCGCG AGCCTCCGGA TGACGACCGT AGTGATGAAT
 3841 CTCTCCTGGC GGGAACAGCA AAATATCACC CGGTCGGCAA ACAAATTCTC GTCCCTGATT
 3901 TTTCACCACC CCCTGACCGC GAATGGTGAG ATTGAGAATA TAACCTTTCA TTCCCAGCGG
 3961 TCGGTCGATA AAAAAATCGA GATAACCGTT GGCCTCAATC GGCGTTAAAC CCGCCACCAG
 4021 ATGGGCATTA AACGAGTATC CCGGCAGCAG GGGATCATTT TGCGCTTCAG CCATACTTTT
 4081 CATACTCCCG CCATTCAGAG
//

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