pET-44a(+)

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pET44a载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET44a, pET 44a
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: 7311 bp
5' 测序引物序列:   T7: 5'-TAATACGACTCACTATAGGG-3'
3' 测序引物序列:   ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3'
载体标签: C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK
载体抗性: 氨苄
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
备注:
载体同时含有N端及C端His标签; 含有N端Thrombin和EK蛋白酶酶切位点; pET44a,b,c 载体的差异主要是在多克隆位点.
产品目录号: 71122-3
稳定性: 瞬时表达
组成型: 组成型
病毒/非病毒: 非病毒

pET44a载体质粒图谱和多克隆位点信息

pet44a载体图谱
pet44a多克隆位点

pET44a载体简介

pET44系列载体是设计用来克隆和高水平表达含有495个氨基酸的Nus标签的蛋白表达载体。与pET-43.1系列载体相比,pET44系列载体额外编码一个N端的His蛋白纯化标签。
pET42a载体的单一的多克隆位点见上面的环状质粒图谱。注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面 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。对pET44a载体的单链测序可以使用COLIDOWN primer(Cat. No. 70845-3).<
把表达的基因插入到载体的PshAI或者SmaI限制性内切酶位点时,可以通过EK蛋白酶或者Thrombin蛋白酶将融合表达的Nus标签完全切除。
pET Vectors 选择指南

pET44a载体序列

LOCUS       pET-44 a(+)    7311 bp     DNA   circular  SYN
DEFINITION  pET-44 a(+)
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..7311
                     /organism="pET-44 a(+)"
                     /mol_type="other DNA"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     terminator      complement(141..168)
                     /label="rrnB_T2_terminator"
     terminator      complement(300..343)
                     /label="rrnB_T1_terminator"
     terminator      complement(309..466)
                     /label="rrnB_terminator"
     CDS             complement(549..2354)
                     /label="ORF frame 2"
     CDS             618..2132
                     /label="ORF frame 3"
     misc_feature    complement(681..695)
                     /label="EK" 
     misc_feature    complement(747..791)
                     /label="S15"  
     CDS             2325..3083
                     /label="ORF frame 3"
     misc_feature    complement(2325..2342)
                     /label="6xHIS"  
     misc_feature    complement(2363..2379)
                     /label="T7_transl_en_RBS" 
     misc_feature    complement(2397..2424)
                     /label="lacO"  
     promoter        complement(2424..2442)
                     /label="T7_promoter"   
     gene            2478..2741
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"  
     misc_feature    complement(2530..2549)
                     /label="pBRrevBam_primer" 
     misc_feature    2824..3915
                     /label="lacI"    
     CDS             2956..3915
                     /label="ORF frame 1" 
     misc_feature    4487..4678
                     /label="ROP"    
     misc_feature    complement(4672..4694)
                     /label="pGEX_3_primer"  
     rep_origin      complement(5093..5712)
                     /label="pBR322_origin"    
     promoter        5799..5827
                     /label="AmpR_promoter"  
     gene            5870..6730
                     /label="Ampicillin"
                     /gene="Ampicillin"  
     CDS             5870..6730
                     /label="ORF frame 2" 
ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggata catatttgaa tgtatttaga aaaataaaca
  121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
  181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
  241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
  301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
  361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
  421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
  481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
  541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
  601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
  661 gatccacgag ctcccggact cttgtcgtcg tcatcaatcg taccagaacc cgcgctgccc
  721 cgggggacca gaccagttgg cggtggcgag tccatgtgct ggcgttcaaa tttcgcagca
  781 gcggtttctt tacccgcgga gtgatggtga tggtgatgac cagaaccact agtcgcttcg
  841 tcaccgaacc agcaaatatt acgggcagcc ataatcagtg ctccggcttt ttcgtcggtc
  901 aacccttcga tatcagccag atcatcaatg ccctgttcgg cgagatcttc cagcgtacaa
  961 acgccacggg cggccagttt gaatgccaaa tcacgatcta ccccttcaag gttcagcaga
 1021 tcgtcagccg gtttgttatc accgaggctt tcttcctggg cctgtgcaat ggtggccagt
 1081 gcatttttag cacgctcgcg cagtgcttca acggtcggct catcaaggcc ttcgatttcc
 1141 aacagctctt tcatcggcac ataggccaat tcttccagcg tcgagaagcc ttcttctacc
 1201 agaacagtcg cgaagtcttc gtcgatgtcg agatatttgg tgaaggtgtc gatcgctgcg
 1261 tgcgcttccg cctgatgctt agcttgcagg tcgtcaacgg tcatcacgtt gagttcccaa
 1321 ccgctcagtt gcgaagccag acgcacgttc tgaccgttac ggccaatcgc ctgcgccaga
 1381 ttaccggctt caacggcgat gtccatggtg tgtttatctt catccaccac gatagaagca
 1441 acgtctgccg gtgccattgc gttaatcacg aactgcgccg ggttatcatc ccacaggacg
 1501 atatcgatac gctcgccacc cagttcagta gacaccgcct gaacacgcgc gccacgcata
 1561 cctacgcaag cacctaccgg atcgatacgt ttatcgttgg ttttcaccgc gattttcgca
 1621 cgagaacccg gatcgcgagc cgctgcttta atttcaatca cttcttcgcc gatttctggc
 1681 acttcaatac ggaacagttc gatcagcatt tccggcttgg aacgagtgac gaacagttgc
 1741 gcgccacgcg cttccgggcg aacggaatag agcacgccac gaacgcggtc gccagggcgg
 1801 aagttttcac gcggcagcat atcttcgcgc aggatcacgg cttcagcgtt gttgcccaga
 1861 tccagagaga tgttgtcgcg gtttactttt ttcaccacgc cggtgatgat ttcaccttcg
 1921 tgttcacgga actgatcaac caccatcgca cgttcggctt cacgcacttt ctgcacgata
 1981 acctgttttg ccgtctgggt agtgatacgg tcaaaggtaa cagactcaat ctgatcttca
 2041 acgtaatcgc ccaggttcag gctttcatct tcataacgtg cggcttcaag ggtgatttcc
 2101 ttggtcggct gggtgacttc atcaacaact aaccagcgac ggaaagtgtc aaaatcaccg
 2161 cttttgcgat cgatctgtac gcggacgtcg atctcttgtt catatttttt ctttgttgct
 2221 gtcgccagcg cgctttccaa tgcttcgaaa atcttctcgc gaggtagcgc cttttcattg
 2281 gatacggctt caactacagc caaaatttct ttgttcatac tagaatggtg atggtggtga
 2341 tggctgctgc ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
 2401 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
 2461 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
 2521 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
 2581 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
 2641 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
 2701 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
 2761 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
 2821 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
 2881 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
 2941 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
 3001 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
 3061 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
 3121 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
 3181 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
 3241 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
 3301 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
 3361 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
 3421 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
 3481 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
 3541 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
 3601 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc
 3661 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
 3721 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
 3781 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
 3841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
 3901 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
 3961 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
 4021 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
 4081 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
 4141 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
 4201 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
 4261 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
 4321 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
 4381 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
 4441 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
 4501 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
 4561 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
 4621 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
 4681 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
 4741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
 4801 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
 4861 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
 4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
 4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
 5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
 5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
 5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
 5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
 5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
 5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
 5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
 5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
 5521 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
 5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
 5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
 5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
 5761 ttttggtcat gacctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
 5821 agacaataac cctgataaat gcttcaatca tgattgaaaa aggaagagta tgagtattca
 5881 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
 5941 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
 6001 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
 6061 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
 6121 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
 6181 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
 6241 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
 6301 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
 6361 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
 6421 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
 6481 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
 6541 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
 6601 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
 6661 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
 6721 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
 6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatgc catcgtgccg
 6841 ccagaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
 6901 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
 6961 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
 7021 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
 7081 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
 7141 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
 7201 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
 7261 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a

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pET44a载体质粒图谱pdf版和相关资料下载

pET44a载体应用举例

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