pET-44a(+)
pET-44a(+)
编号 | 载体名称 |
北京华越洋生物VECT5250 | pET-44a(+) |
pET44a载体基本信息
别名: | pET44a, pET 44a |
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: | 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 |
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. |
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. 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。
pET44a载体序列
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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其他大肠杆菌表达载体: