pET-44 EK/LIC

pET-44 EK/LIC

pET44 EK/LIC载体基本信息

出品公司:	Ybscience
别名:	pET44 EK/LIC, pET 44 EK/LIC
质粒类型:	大肠杆菌蛋白表达
表达水平:	高
克隆方法:	多克隆位点，限制性内切酶
载体大小:	7331 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
备注:	For directional cloning of PCR-amplified DNA; ligation independent cloning; enterokinase cleavage site.
稳定性:	瞬时表达 Transient
表达宿主菌:	BL21(DE3) 、 Rosetta2(DE3)、C43(DE3)
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pET44 EK/LIC载体质粒图谱和多克隆位点信息

 

pET44 EK/LIC载体简介

The pET-44 Ek/LIC Vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides fused with N-terminal His•Tag®,Nus•Tag™ and S•Tag™ sequences. Using specifically designed primers for amplification and the pET-44 Ek/LIC Vector Kit (Cat. No. 71144-3), 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 sequences can be completely removed when cloning into the Ek/LIC site (as shown below left) by cleaving the fusion protein with enterokinase.

pET44 EK/LIC载体序列

LOCUS       pET-44 Ek/LIC    7331 bp     DNA Circular    SYN
DEFINITION  pET-44 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..7331
                     /organism="pET-44 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..2374)
                     /label="ORF frame 2"
    misc_feature    complement(701..715)
                     /label="EK" 
     misc_feature    complement(767..811)
                     /label="S15"  
     CDS             824..2152
                     /label="ORF frame 2"  
     CDS             2345..3103
                     /label="ORF frame 2"   
     misc_feature    complement(2345..2362)
                     /label="6xHIS"
     misc_feature    complement(2383..2399)
                     /label="T7_transl_en_RBS"     
     misc_feature    complement(2417..2444)
                     /label="lacO"       
     promoter        complement(2444..2462)
                     /label="T7_promoter"  
     gene            2498..2761
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"  
     misc_feature    complement(2550..2569)
                     /label="pBRrevBam_primer"    
     misc_feature    2844..3935
                     /label="lacI"
     CDS             2976..3935
                     /label="ORF frame 3"
     misc_feature    4507..4698
                     /label="ROP" 
     misc_feature    complement(4692..4714)
                     /label="pGEX_3_primer"    
     rep_origin      complement(5113..5732)
                     /label="pBR322_origin"   
     promoter        5819..5847
                     /label="AmpR_promoter"
     gene            5890..6750
                     /label="Ampicillin"
                     /gene="Ampicillin"      
     CDS             5890..6750
                     /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 ttgttcatac
 2341 tagaatggtg atggtggtga tggctgctgc ccatatgtat atctccttct taaagttaaa
 2401 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
 2461 taatttcgcg ggatcgagat cgatctcgat cctctacgcc ggacgcatcg tggccggcat
 2521 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
 2581 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
 2641 cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt
 2701 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga
 2761 gcgtcgagat cccggacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
 2821 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
 2881 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
 2941 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
 3001 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
 3061 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
 3121 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
 3181 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
 3241 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
 3301 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
 3361 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
 3421 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
 3481 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
 3541 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
 3601 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg
 3661 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
 3721 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
 3781 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
 3841 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
 3901 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
 3961 ctcactcatt aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc
 4021 tccttccggt gggcgcgggg catgactagc atgatcgtgc tcctgtcgtt gaggacccgg
 4081 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
 4141 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
 4201 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
 4261 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
 4321 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
 4381 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
 4441 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
 4501 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
 4561 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
 4621 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
 4681 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
 4741 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
 4801 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
 4861 cagagcagat tgtactgaga gtgcaccata tatgcggtgt gaaataccgc acagatgcgt
 4921 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
 4981 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
 5041 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
 5101 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
 5161 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
 5221 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
 5281 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
 5341 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
 5401 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
 5461 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
 5521 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
 5581 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
 5641 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
 5701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
 5761 cgaaaactca cgttaaggga ttttggtcat gacctatttg tttatttttc taaatacatt
 5821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaatca tgattgaaaa
 5881 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
 5941 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
 6001 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
 6061 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
 6121 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
 6181 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
 6241 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
 6301 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
 6361 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
 6421 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
 6481 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
 6541 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
 6601 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
 6661 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
 6721 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
 6781 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
 6841 aatctcatgc catcgtgccg ccagaaattg taaacgttaa tattttgtta aaattcgcgt
 6901 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
 6961 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
 7021 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
 7081 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
 7141 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
 7201 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
 7261 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
 7321 cccattcgcc a
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