pET-45b(+)

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pET45b载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET45b, pET 45b
质粒类型: 大肠杆菌蛋白表达
表达水平:
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5260 bp
5' 测序引物: T7
5' 测序引物序列:   T7: 5'-TAATACGACTCACTATAGGG-3'
3' 测序引物: T7t
3' 测序引物序列:   T7t: 5'-TGCTAGTTATTGCTCAGCGG-3'
载体标签: N-His,N-EK, C-S
载体抗性: Ampicillin
备注:
N-term HisTag with minimal extraneous sequences.
产品目录号: 71327-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒
表达宿主菌: BL21(DE3)BLR(DE3)Rosetta(DE3)OrigamiB(DE3)C43(DE3)
pET表达手册: pET表达手册英文版
pET表达手册中文版
pET载体汇总:
pET-3a     pET-11a   pET-12a   pET-14b   pET-15b   pET-16b
pET-17b   pET-19b   pET-20b   pET-21a   pET-22b   pET-23a
pET-24a   pET-25b   pET-26b   pET-27b   pET-28a   pET-29a
pET-30a   pET-31b   pET-32a   pET-33b   pET-34b   pET-35b
pET-37b   pET-39b   pET-40b   pET-41a   pET-42a   pET-43.1a
pET-44a   pET-45b   pET-47b   pET-48b   pET-49b   pET-50b
pET-51b   pET-52b  
pETDuet-1  pACYCDuet-1  pCDFDuet-1 pRSFDuet-1  pColaDuet-1

pET45b载体图谱质粒图谱和多克隆位点信息

pet45b载体图谱
pet45b多克隆位点

Feature Name Start End  
Ampicillin 1442 582
AmpR_promoter 1513 1485
pBR322_origin 1600 2219
pGEX_3_primer 2618 2640
ROP 2825 2634
lacI 4488 3397
tet (887 - 636) 4831 4580
pBRrevBam_primer 4763 4782
T7_promoter 4870 4888
lacO 4888 4915
T7_transl_en_RBS 4933 4949
EK 4999 5013
T7_Terminal_primer 5192 5174
T7_terminator 5193 5228
ORF Start End  
ORF frame 3 1442 582
ORF frame 2 4356 3397
ORF frame 1 4729 4229
Enzyme Name Cut
FspI 877
HpaI 3622
ApaI 3921
BclI 4110
ClaI 4849
XbaI 4916
NcoI 4955
PmlI 4980
KpnI 4988
AgeI 4987
BstBI 4993
BamHI 5019
SacI 5036
AscI 5047
PstI 5057
SalI 5059
HindIII 5065
NotI 5072
EagI 5072
XhoI 5080
PacI 5155

pET45b载体简介

pET-45b载体是带有N-His蛋白标签,EK蛋白酶位点和C-S标签,可以在大肠杆菌中高表达目的蛋白。独特的酶切位点在载体图谱中被标注出来。需要注意的是,这些载体序列是依据载体pBR322的方向来进行编号的,所以T7标签表达区域在载体图谱上是反向的。

T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的,所以在T7噬菌体聚合酶的作用下,包含有蛋白编码序列的病毒 粒子能够产生,并启动蛋白表达,同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。   pET-45b载体上面含有一个EK蛋白酶切位点,当将目的基因使用载体上面的PshAI限制性内切酶位点插入进去时,可以通过EK蛋白酶将载体上的融合的所有氨基酸序列包括标签序列,全部切除掉。



pET Vectors 选择指南

pET45b载体序列

LOCUS       pET-45b(+)    5260 bp     DNA  Circular  SYN
DEFINITION  pET-45b(+)
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTOAUTHORAD1|Novagen Inc.|
FEATURES             Location/Qualifiers
     source          1..5260
                     /organism="pET-45b(+)"
                     /mol_type="other DNA"
     gene            complement(582..1442)
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             complement(582..1442)
                     /label="ORF frame 3"    
     promoter        complement(1485..1513)
                     /label="AmpR_promoter"   
     rep_origin      1600..2219
                     /label="pBR322_origin"  
     misc_feature    2618..2640
                     /label="pGEX_3_primer"      
     misc_feature    complement(2634..2825)
                     /label="ROP"     
     misc_feature    complement(3397..4488)
                     /label="lacI"       
     CDS             complement(3397..4356)
                     /label="ORF frame 2"       
     CDS             complement(4229..4729)
                     /label="ORF frame 1"      
     gene            complement(4580..4831)
                     /label="tet (887 - 636)"
                     /gene="tet (887 - 636)"         
     misc_feature    4763..4782
                     /label="pBRrevBam_primer"            
     promoter        4870..4888
                     /label="T7_promoter"         
     misc_feature    4888..4915
                     /label="lacO"       
     misc_feature    4933..4949
                     /label="T7_transl_en_RBS"            
     misc_feature    4999..5013
                     /label="EK"                
     misc_feature    complement(5174..5192)
                     /label="T7_Terminal_primer"                  
     terminator      5193..5228
                     /label="T7_terminator"             
ORIGIN
    1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
   61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
  121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
  181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
  241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
  301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
  361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
  421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat
  481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
  541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
  601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
  661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
  721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
  781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
  841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
  901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
  961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
 1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
 1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
 1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
 1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
 1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
 1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
 1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
 1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
 1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
 1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
 1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
 1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
 1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
 1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
 1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
 1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
 1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
 2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
 2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
 2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
 2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
 2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
 2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
 2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
 2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
 2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
 2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
 2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
 2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
 2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
 2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
 2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
 2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
 3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
 3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
 3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
 3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
 3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
 3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
 3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
 3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
 3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
 3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
 3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
 3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
 3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
 3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
 3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
 3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
 3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
 4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
 4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
 4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
 4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
 4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
 4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
 4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
 4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
 4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
 4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
 4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
 4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
 4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
 4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
 4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
 4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac
 4981 gtgggtaccg gttcgaatga tgacgacgac aagagtccgg atcccaattg ggagctcgtg
 5041 tacacggcgc gcctgcaggt cgacaagctt gcggccgcac tcgagtctgg taaagaaacc
 5101 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc
 5161 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
 5221 ttgaggggtt ttttgctgaa aggaggaact atatccggat
//

pET45b载体图谱质粒图谱pdf版和相关资料下载

pET45b载体质粒应用举例

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