pET-43.1 EK/LIC

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pET43.1 EK/LIC载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET43.1 EK/LIC, pET 43.1 EK/LIC
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: 7295 bp
5' 测序引物: T7
5' 测序引物序列:   T7: 5'-TAATACGACTCACTATAGGG-3'
3' 测序引物: ColiDOWN
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: Ampicillin
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
备注:
For directional cloning of PCR-amplified DNA; ligation independent cloning; enterokinase cleavage site
产品目录号: 71072-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pET43.1 EK/LIC载体质粒图谱和多克隆位点信息

pet43.1 EK/LIC载体图谱
pet43.1 EK/LIC多克隆位点
pet43.1 EK/LIC载体克隆方法说明

pET43.1 EK/LIC载体简介

The pET-43.1 Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides fused with N-terminal Nus•Tag™, His•Tag® and S•Tag™ sequences. Using specifically designed primers for amplification and the pET-43.1 Ek/LIC Vector Kit (Cat. No. 71072-3), inserts can be efficiently cloned without the need for restriction digestion or ligation. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the COLIDOWN primer (Cat. No. 70845-3). Vector encoded sequence can be completely removed when cloning into the Ek/LIC site (as shown below left) by cleaving the fusion protein with enterokinase.

pET43.1 EK/LIC载体序列

LOCUS       pET-43.1 Ek/LIC    7295 bp     DNA Circular    SYN
DEFINITION  pET-43.1 Ek/LIC
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..7295
                     /organism="pET-43.1 Ek/LIC"
                     /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(509..2338)
                     /label="ORF frame 2"
      misc_feature    complement(701..715)
                     /label="EK"
      misc_feature    complement(767..811)
                     /label="S15"
     misc_feature    complement(821..838)
                     /label="6xHIS"
     CDS             824..2152
                     /label="ORF frame 2" 
     CDS             2339..3067
                     /label="ORF frame 2" 
     misc_feature    complement(2347..2363)
                     /label="T7_transl_en_RBS"  
     misc_feature    complement(2381..2408)
                     /label="lacO"   
     promoter        complement(2408..2426)
                     /label="T7_promoter"
     gene            2462..2725
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(2514..2533)
                     /label="pBRrevBam_primer"
     misc_feature    2808..3899
                     /label="lacI"   
     CDS             2940..3899
                     /label="ORF frame 3"
     misc_feature    4471..4662
                     /label="ROP" 
     misc_feature    complement(4656..4678)
                     /label="pGEX_3_primer" 
     rep_origin      complement(5077..5696)
                     /label="pBR322_origin" 
     promoter        5783..5811
                     /label="AmpR_promoter" 
     gene            5854..6714
                     /label="Ampicillin"
                     /gene="Ampicillin"  
     CDS             5854..6714
                     /label="ORF frame 1"
      
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 ctcagatatc gttgaggaga agcccggact cttgtcgtcg tcatcaatcg
  721 taccagaacc cgcgctgccc cgggggacca gaccagttgg cggtggcgag tccatgtgct
  781 ggcgttcaaa tttcgcagca gcggtttctt tacccgcgga gtgatggtga tggtgatgac
  841 cagaaccact agtcgcttcg tcaccgaacc agcaaatatt acgggcagcc ataatcagtg
  901 ctccggcttt ttcgtcggtc aacccttcga tatcagccag atcatcaatg ccctgttcgg
  961 cgagatcttc cagcgtacaa acgccacggg cggccagttt gaatgccaaa tcacgatcta
 1021 ccccttcaag gttcagcaga tcgtcagccg gtttgttatc accgaggctt tcttcctggg
 1081 cctgtgcaat ggtggccagt gcatttttag cacgctcgcg cagtgcttca acggtcggct
 1141 catcaaggcc ttcgatttcc aacagctctt tcatcggcac ataggccaat tcttccagcg
 1201 tcgagaagcc ttcttctacc agaacagtcg cgaagtcttc gtcgatgtcg agatatttgg
 1261 tgaaggtgtc gatcgctgcg tgcgcttccg cctgatgctt agcttgcagg tcgtcaacgg
 1321 tcatcacgtt gagttcccaa ccgctcagtt gcgaagccag acgcacgttc tgaccgttac
 1381 ggccaatcgc ctgcgccaga ttaccggctt caacggcgat gtccatggtg tgtttatctt
 1441 catccaccac gatagaagca acgtctgccg gtgccattgc gttaatcacg aactgcgccg
 1501 ggttatcatc ccacaggacg atatcgatac gctcgccacc cagttcagta gacaccgcct
 1561 gaacacgcgc gccacgcata cctacgcaag cacctaccgg atcgatacgt ttatcgttgg
 1621 ttttcaccgc gattttcgca cgagaacccg gatcgcgagc cgctgcttta atttcaatca
 1681 cttcttcgcc gatttctggc acttcaatac ggaacagttc gatcagcatt tccggcttgg
 1741 aacgagtgac gaacagttgc gcgccacgcg cttccgggcg aacggaatag agcacgccac
 1801 gaacgcggtc gccagggcgg aagttttcac gcggcagcat atcttcgcgc aggatcacgg
 1861 cttcagcgtt gttgcccaga tccagagaga tgttgtcgcg gtttactttt ttcaccacgc
 1921 cggtgatgat ttcaccttcg tgttcacgga actgatcaac caccatcgca cgttcggctt
 1981 cacgcacttt ctgcacgata acctgttttg ccgtctgggt agtgatacgg tcaaaggtaa
 2041 cagactcaat ctgatcttca acgtaatcgc ccaggttcag gctttcatct tcataacgtg
 2101 cggcttcaag ggtgatttcc ttggtcggct gggtgacttc atcaacaact aaccagcgac
 2161 ggaaagtgtc aaaatcaccg cttttgcgat cgatctgtac gcggacgtcg atctcttgtt
 2221 catatttttt ctttgttgct gtcgccagcg cgctttccaa tgcttcgaaa atcttctcgc
 2281 gaggtagcgc cttttcattg gatacggctt caactacagc caaaatttct ttgttcatat
 2341 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
 2401 caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta
 2461 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
 2521 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
 2581 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
 2641 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
 2701 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
 2761 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
 2821 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
 2881 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
 2941 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
 3001 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
 3061 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
 3121 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
 3181 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
 3241 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
 3301 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
 3361 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
 3421 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
 3481 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
 3541 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
 3601 tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
 3661 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
 3721 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
 3781 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
 3841 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
 3901 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
 3961 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tagcatgatc
 4021 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
 4081 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
 4141 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca
 4201 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
 4261 acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc
 4321 cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca
 4381 tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg
 4441 aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac
 4501 atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac
 4561 gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc
 4621 agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
 4681 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
 4741 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
 4801 tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatatgcg
 4861 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
 4921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
 4981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
 5041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
 5101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
 5161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
 5221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
 5281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
 5341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
 5401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
 5461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
 5521 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
 5581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
 5641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
 5701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaccta
 5761 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
 5821 aaatgcttca atcatgattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
 5881 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
 5941 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
 6001 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
 6061 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
 6121 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
 6181 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
 6241 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
 6301 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
 6361 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
 6421 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
 6481 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
 6541 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
 6601 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
 6661 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
 6721 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
 6781 atctaggtga agatcctttt tgataatctc atgccatcgt gccgccagaa attgtaaacg
 6841 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
 6901 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
 6961 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
 7021 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
 7081 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
 7141 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
 7201 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
 7261 ttaatgcgcc gctacagggc gcgtcccatt cgcca
//

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pET43.1 EK/LIC载体应用举例

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