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pET-45b(+)

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pET-45b(+)載體 pET45b質粒基本信息

出品公司: EMD Biosciences (Novagen)
別名: pET45b, pET 45b
質粒類型: 大腸桿菌蛋白表達
表達水平:
克隆方法: 多克隆位點,限制性內切酶
載體大小: 5260 bp
5' 測序引物: T7
5' 測序引物序列:   T7: 5'-TAATACGACTCACTATAGGG-3'
3' 測序引物: T7t
3' 測序引物序列:   T7t: 5'-TGCTAGTTATTGCTCAGCGG-3'
載體標簽: N-His,N-EK, C-S
載體抗性: Ampicillin
備注:
N-term HisTag with minimal extraneous sequences.
產品目錄號: 71327-3
穩定性: 瞬時表達 Transient
組成型: 組成型 Constitutive
病毒/非病毒: 非病毒
表達宿主菌: BL21(DE3)BLR(DE3)Rosetta(DE3)OrigamiB(DE3)C43(DE3)
pET表達手冊: pET表達手冊英文版
pET表達手冊中文版
pET載體匯總:
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

pET-45b(+)質粒圖譜 pET45b載體圖譜和pET-45b(+)載體序列 pET45b質粒序列多克隆位點信息

pet45b載體圖譜
pet45b多克隆位點

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

pET-45b(+)質粒 pET45b載體簡介

pET-45b載體是帶有N-His蛋白標簽,EK蛋白酶位點和C-S標簽,可以在大腸桿菌中高表達目的蛋白。獨特的酶切位點在載體圖譜中被標注出來。需要注意的是,這些載體序列是依據載體pBR322的方向來進行編號的,所以T7標簽表達區域在載體圖譜上是反向的。

T7 RNA聚合酶啟動的克隆和表達區域在質粒圖譜中也被標注了出來。質粒的F1復制子是被定向的,所以在T7噬菌體聚合酶的作用下,包含有蛋白編碼序列的病毒 粒子能夠產生,并啟動蛋白表達,同時蛋白表達將被T7終止子序列(Cat. No. 69337-3)的作用下終止蛋白翻譯。   pET-45b載體上面含有一個EK蛋白酶切位點,當將目的基因使用載體上面的PshAI限制性內切酶位點插入進去時,可以通過EK蛋白酶將載體上的融合的所有氨基酸序列包括標簽序列,全部切除掉。



pET Vectors 選擇指南

pET-45b(+)質粒序列 pET45b載體序列

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
            //gt12365.cn/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|gt12365.cn|
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
//

pET-45b(+)載體圖譜 pET45b質粒圖譜pdf版和pET-45b(+)載體序列 pET45b質粒序列等相關資料下載

pET-45b(+)質粒 pET45b載體應用舉例

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