| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 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.
|
| 产品目录号: | 71144-3 |
| 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 |
| 表达宿主菌2: | BLR(DE3)、Rosetta(DE3)、OrigamiB(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 |
买家导航
pET-44 EK/LIC载体pET44 EK/LIC质粒基本信息
pET-44 EK/LIC质粒图谱pET44 EK/LIC载体图谱和pET-44 EK/LIC载体序列pET44 EK/LIC质粒序列多克隆位点信息
pET-44 EK/LIC质粒pET44 EK/LIC载体简介
pET-44 EK/LIC载体是用来快速定向PCR高通量质粒构建的载体。本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-c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pET44 EK LIC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-44 EK/LIC质粒序列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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pET-44 EK/LIC质粒pET44 EK/LIC载体应用举例
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