pBad/Myc-His A

pBad/Myc-His A

pBadMyc-His A载体基本信息

出品公司:	Ybscience
载体名称:	pBAD/Myc-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:	4094 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（C-端）,c-Myc Epitope Tag（C-端）
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/Myc-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；
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

 

pBadMyc-His A载体质粒图谱和多克隆位点信息

pBadMyc-His 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/Myc-His A,B,C 载体简介

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 AntibodyThree 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.

 

pBadMyc-His A载体序列

LOCUS       pBAD/myc-His A	4094 bp 	DNA	circular	SYN
DEFINITION  pBAD/myc-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..4094
                     /organism="pBAD/myc-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    complement(495..512)
                     /label="pTrcHis_rev_primer"
     misc_feature    complement(495..512)
                     /label="pBAD_rev_primer"
     terminator      545..702
                     /label="rrnB_terminator"
     terminator      668..711
                     /label="rrnB_T1_terminator"
     terminator      843..870
                     /label="rrnB_T2_terminator"
     promoter        911..939
                     /label="AmpR_promoter"
     gene            981..1841
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             981..1841
                     /label="ORF frame 3"                     
     rep_origin      1996..2615
                     /label="pBR322_origin"                     
     misc_feature    3012..3034
                     /label="pGEX_3_primer"                     
     misc_feature    complement(3190..4068)
                     /label="AraC"                     
     CDS             complement(3190..25)
                     /label="ORF frame 2"                     
     CDS             complement(3190..4077)
                     /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 GGGCCCGAAC AAAAACTCAT CTCAGAAGAG GATCTGAATA GCGCCGTCGA
  421 CCATCATCAT CATCATCATT GAGTTTAAAC GGTCTCCAGC TTGGCTGTTT TGGCGGATGA
  481 GAGAAGATTT TCAGCCTGAT ACAGATTAAA TCAGAACGCA GAAGCGGTCT GATAAAACAG
  541 AATTTGCCTG GCGGCAGTAG CGCGGTGGTC CCACCTGACC CCATGCCGAA CTCAGAAGTG
  601 AAACGCCGTA GCGCCGATGG TAGTGTGGGG TCTCCCCATG CGAGAGTAGG GAACTGCCAG
  661 GCATCAAATA AAACGAAAGG CTCAGTCGAA AGACTGGGCC TTTCGTTTTA TCTGTTGTTT
  721 GTCGGTGAAC GCTCTCCTGA GTAGGACAAA TCCGCCGGGA GCGGATTTGA ACGTTGCGAA
  781 GCAACGGCCC GGAGGGTGGC GGGCAGGACG CCCGCCATAA ACTGCCAGGC ATCAAATTAA
  841 GCAGAAGGCC ATCCTGACGG ATGGCCTTTT TGCGTTTCTA CAAACTCTTT TGTTTATTTT
  901 TCTAAATACA TTCAAATATG TATCCGCTCA TGAGACAATA ACCCTGATAA ATGCTTCAAT
  961 AATATTGAAA AAGGAAGAGT ATGAGTATTC AACATTTCCG TGTCGCCCTT ATTCCCTTTT
 1021 TTGCGGCATT TTGCCTTCCT GTTTTTGCTC ACCCAGAAAC GCTGGTGAAA GTAAAAGATG
 1081 CTGAAGATCA GTTGGGTGCA CGAGTGGGTT ACATCGAACT GGATCTCAAC AGCGGTAAGA
 1141 TCCTTGAGAG TTTTCGCCCC GAAGAACGTT TTCCAATGAT GAGCACTTTT AAAGTTCTGC
 1201 TATGTGGCGC GGTATTATCC CGTGTTGACG CCGGGCAAGA GCAACTCGGT CGCCGCATAC
 1261 ACTATTCTCA GAATGACTTG GTTGAGTACT CACCAGTCAC AGAAAAGCAT CTTACGGATG
 1321 GCATGACAGT AAGAGAATTA TGCAGTGCTG CCATAACCAT GAGTGATAAC ACTGCGGCCA
 1381 ACTTACTTCT GACAACGATC GGAGGACCGA AGGAGCTAAC CGCTTTTTTG CACAACATGG
 1441 GGGATCATGT AACTCGCCTT GATCGTTGGG AACCGGAGCT GAATGAAGCC ATACCAAACG
 1501 ACGAGCGTGA CACCACGATG CCTGTAGCAA TGGCAACAAC GTTGCGCAAA CTATTAACTG
 1561 GCGAACTACT TACTCTAGCT TCCCGGCAAC AATTAATAGA CTGGATGGAG GCGGATAAAG
 1621 TTGCAGGACC ACTTCTGCGC TCGGCCCTTC CGGCTGGCTG GTTTATTGCT GATAAATCTG
 1681 GAGCCGGTGA GCGTGGGTCT CGCGGTATCA TTGCAGCACT GGGGCCAGAT GGTAAGCCCT
 1741 CCCGTATCGT AGTTATCTAC ACGACGGGGA GTCAGGCAAC TATGGATGAA CGAAATAGAC
 1801 AGATCGCTGA GATAGGTGCC TCACTGATTA AGCATTGGTA ACTGTCAGAC CAAGTTTACT
 1861 CATATATACT TTAGATTGAT TTAAAACTTC ATTTTTAATT TAAAAGGATC TAGGTGAAGA
 1921 TCCTTTTTGA TAATCTCATG ACCAAAATCC CTTAACGTGA GTTTTCGTTC CACTGAGCGT
 1981 CAGACCCCGT AGAAAAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT
 2041 GCTGCTTGCA AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC
 2101 TACCAACTCT TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTCC
 2161 TTCTAGTGTA GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC
 2221 TCGCTCTGCT AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG
 2281 GGTTGGACTC AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT
 2341 CGTGCACACA GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG
 2401 AGCTATGAGA AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG
 2461 GCAGGGTCGG AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT
 2521 ATAGTCCTGT CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG
 2581 GGGGGCGGAG CCTATGGAAA AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT
 2641 GCTGGCCTTT TGCTCACATG TTCTTTCCTG CGTTATCCCC TGATTCTGTG GATAACCGTA
 2701 TTACCGCCTT TGAGTGAGCT GATACCGCTC GCCGCAGCCG AACGACCGAG CGCAGCGAGT
 2761 CAGTGAGCGA GGAAGCGGAA GAGCGCCTGA TGCGGTATTT TCTCCTTACG CATCTGTGCG
 2821 GTATTTCACA CCGCATATGG TGCACTCTCA GTACAATCTG CTCTGATGCC GCATAGTTAA
 2881 GCCAGTATAC ACTCCGCTAT CGCTACGTGA CTGGGTCATG GCTGCGCCCC GACACCCGCC
 2941 AACACCCGCT GACGCGCCCT GACGGGCTTG TCTGCTCCCG GCATCCGCTT ACAGACAAGC
 3001 TGTGACCGTC TCCGGGAGCT GCATGTGTCA GAGGTTTTCA CCGTCATCAC CGAAACGCGC
 3061 GAGGCAGCAG ATCAATTCGC GCGCGAAGGC GAAGCGGCAT GCATAATGTG CCTGTCAAAT
 3121 GGACGAAGCA GGGATTCTGC AAACCCTATG CTACTCCGTC AAGCCGTCAA TTGTCTGATT
 3181 CGTTACCAAT TATGACAACT TGACGGCTAC ATCATTCACT TTTTCTTCAC AACCGGCACG
 3241 GAACTCGCTC GGGCTGGCCC CGGTGCATTT TTTAAATACC CGCGAGAAAT AGAGTTGATC
 3301 GTCAAAACCA ACATTGCGAC CGACGGTGGC GATAGGCATC CGGGTGGTGC TCAAAAGCAG
 3361 CTTCGCCTGG CTGATACGTT GGTCCTCGCG CCAGCTTAAG ACGCTAATCC CTAACTGCTG
 3421 GCGGAAAAGA TGTGACAGAC GCGACGGCGA CAAGCAAACA TGCTGTGCGA CGCTGGCGAT
 3481 ATCAAAATTG CTGTCTGCCA GGTGATCGCT GATGTACTGA CAAGCCTCGC GTACCCGATT
 3541 ATCCATCGGT GGATGGAGCG ACTCGTTAAT CGCTTCCATG CGCCGCAGTA ACAATTGCTC
 3601 AAGCAGATTT ATCGCCAGCA GCTCCGAATA GCGCCCTTCC CCTTGCCCGG CGTTAATGAT
 3661 TTGCCCAAAC AGGTCGCTGA AATGCGGCTG GTGCGCTTCA TCCGGGCGAA AGAACCCCGT
 3721 ATTGGCAAAT ATTGACGGCC AGTTAAGCCA TTCATGCCAG TAGGCGCGCG GACGAAAGTA
 3781 AACCCACTGG TGATACCATT CGCGAGCCTC CGGATGACGA CCGTAGTGAT GAATCTCTCC
 3841 TGGCGGGAAC AGCAAAATAT CACCCGGTCG GCAAACAAAT TCTCGTCCCT GATTTTTCAC
 3901 CACCCCCTGA CCGCGAATGG TGAGATTGAG AATATAACCT TTCATTCCCA GCGGTCGGTC
 3961 GATAAAAAAA TCGAGATAAC CGTTGGCCTC AATCGGCGTT AAACCCGCCA CCAGATGGGC
 4021 ATTAAACGAG TATCCCGGCA GCAGGGGATC ATTTTGCGCT TCAGCCATAC TTTTCATACT
 4081 CCCGCCATTC AGAG