| pPSUMO 大肠杆菌基因表达 SUMO pET-28-SUMO | ||||||||||||||||||||||||
| 发布时间:2018-05-28 12:02:58 | 浏览次数： | ||||||||||||||||||||||||
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标题1测序及更多信息：https://pan.baidu.com/s/1QFW27WjhG3L3BX5mntfjPQ 密码：ntdm采购地址：淘宝店铺地址
载体名称:
pPSUMO
质粒类型:
大肠杆菌基因表达
高拷贝/低拷贝:
高拷贝
启动子:
T7
克隆方法:
多克隆位点，限制性内切酶
载体大小:
5633
bp
5' 测序引物及序列:
T7:
5'-TAATACGACTCACTATAGGG-3'
3' 测序引物及序列:
T7t:
5'-GCTAGTTATTGCTCAGCGG-3'
载体标签:
N-His;
N-SUMO
载体抗性:
Kanamycin 卡那霉素
组成型/诱导型:
诱导型
病毒/非病毒:
非病毒
载体组建方法 This expression vector (termed ppSUMO) was a gift fromP. Pellicini and was originally derived from pET28(Novagen), with the open reading frame for yeast smt3 inserted between the NcoI and BamHI sites. The open reading frame for full-length AAG was PCR-amplified, digested with BamHI and XhoI, and ligated into the ppSUMO vector that had been cut by the same enzymes. The desired construct was verified by sequencing the entire open reading frame of the SUMO fusion. This new expression construct greatly increased the yields of the full-length protein. Baldwin M R, O’Brien P J. Nonspecific DNA binding and coordination of the first two steps of base excision repair[J]. Biochemistry, 2010, 49(36): 7879-7891.
全序列 1 CCCGTCAAGC TCTAAATCGG GGGCTCCCTT TAGGGTTCCG ATTTAGTGCT TTACGGCACC 61 TCGACCCCAA AAAACTTGAT TAGGGTGATG GTTCACGTAG TGGGCCATCG CCCTGATAGA121 CGGTTTTTCG CCCTTTGACG TTGGAGTCCA CGTTCTTTAA TAGTGGACTC TTGTTCCAAA 181 CTGGAACAAC ACTCAACCCT ATCTCGGTCT ATTCTTTTGA TTTATAAGGG ATTTTGCCGA 241 TTTCGGCCTA TTGGTTAAAA AATGAGCTGA TTTAACAAAA ATTTAACGCG AATTTTAACA 301 AAATATTAAC GTTTACAATT TCAGGTGGCA CTTTTCGGGG AAATGTGCGC GGAACCCCTA 361 TTTGTTTATT TTTCTAAATA CATTCAAATA TGTATCCGCT CATGAATTAA TTCTTAGAAA 421 AACTCATCGA GCATCAAATG AAACTGCAAT TTATTCATAT CAGGATTATC AATACCATAT 481 TTTTGAAAAA GCCGTTTCTG TAATGAAGGA GAAAACTCAC CGAGGCAGTT CCATAGGATG 541 GCAAGATCCT GGTATCGGTC TGCGATTCCG ACTCGTCCAA CATCAATACA ACCTATTAAT 601 TTCCCCTCGT CAAAAATAAG GTTATCAAGT GAGAAATCAC CATGAGTGAC GACTGAATCC 661 GGTGAGAATG GCAAAAGTTT ATGCATTTCT TTCCAGACTT GTTCAACAGG CCAGCCATTA 721 CGCTCGTCAT CAAAATCACT CGCATCAACC AAACCGTTAT TCATTCGTGA TTGCGCCTGA 781 GCGAGACGAA ATACGCGATC GCTGTTAAAA GGACAATTAC AAACAGGAAT CGAATGCAAC 841 CGGCGCAGGA ACACTGCCAG CGCATCAACA ATATTTTCAC CTGAATCAGG ATATTCTTCT 901 AATACCTGGA ATGCTGTTTT CCCGGGGATC GCAGTGGTGA GTAACCATGC ATCATCAGGA 961 GTACGGATAA AATGCTTGAT GGTCGGAAGA GGCATAAATT CCGTCAGCCA GTTTAGTCTG 1021 ACCATCTCAT CTGTAACATC ATTGGCAACG CTACCTTTGC CATGTTTCAG AAACAACTCT 1081 GGCGCATCGG GCTTCCCATA CAATCGATAG ATTGTCGCAC CTGATTGCCC GACATTATCG 1141 CGAGCCCATT TATACCCATA TAAATCAGCA TCCATGTTGG AATTTAATCG CGGCCTAGAG 1201 CAAGACGTTT CCCGTTGAAT ATGGCTCATA ACACCCCTTG TATTACTGTT TATGTAAGCA 1261 GACAGTTTTA TTGTTCATGA CCAAAATCCC TTAACGTGAG TTTTCGTTCC ACTGAGCGTC 1321 AGACCCCGTA GAAAAGATCA AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG 1381 CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT 1441 ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTCCT 1501 TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT 1561 CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG 1621 GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC 1681 GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA 1741 GCTATGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG 1801 CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA 1861 TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG 1921 GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG 1981 CTGGCCTTTT GCTCACATGT TCTTTCCTGC GTTATCCCCT GATTCTGTGG ATAACCGTAT 2041 TACCGCCTTT GAGTGAGCTG ATACCGCTCG CCGCAGCCGA ACGACCGAGC GCAGCGAGTC 2101 AGTGAGCGAG GAAGCGGAAG AGCGCCTGAT GCGGTATTTT CTCCTTACGC ATCTGTGCGG 2161 TATTTCACAC CGCATATATG GTGCACTCTC AGTACAATCT GCTCTGATGC CGCATAGTTA 2221 AGCCAGTATA CACTCCGCTA TCGCTACGTG ACTGGGTCAT GGCTGCGCCC CGACACCCGC 2281 CAACACCCGC TGACGCGCCC TGACGGGCTT GTCTGCTCCC GGCATCCGCT TACAGACAAG 2341 CTGTGACCGT CTCCGGGAGC TGCATGTGTC AGAGGTTTTC ACCGTCATCA CCGAAACGCG 2401 CGAGGCAGCT GCGGTAAAGC TCATCAGCGT GGTCGTGAAG CGATTCACAG ATGTCTGCCT 2461 GTTCATCCGC GTCCAGCTCG TTGAGTTTCT CCAGAAGCGT TAATGTCTGG CTTCTGATAA 2521 AGCGGGCCAT GTTAAGGGCG GTTTTTTCCT GTTTGGTCAC TGATGCCTCC GTGTAAGGGG 2581 GATTTCTGTT CATGGGGGTA ATGATACCGA TGAAACGAGA GAGGATGCTC ACGATACGGG 2641 TTACTGATGA TGAACATGCC CGGTTACTGG AACGTTGTGA GGGTAAACAA CTGGCGGTAT 2701 GGATGCGGCG GGACCAGAGA AAAATCACTC AGGGTCAATG CCAGCGCTTC GTTAATACAG 2761 ATGTAGGTGT TCCACAGGGT AGCCAGCAGC ATCCTGCGAT GCAGATCCGG AACATAATGG 2821 TGCAGGGCGC TGACTTCCGC GTTTCCAGAC TTTACGAAAC ACGGAAACCG AAGACCATTC 2881 ATGTTGTTGC TCAGGTCGCA GACGTTTTGC AGCAGCAGTC GCTTCACGTT CGCTCGCGTA 2941 TCGGTGATTC ATTCTGCTAA CCAGTAAGGC AACCCCGCCA GCCTAGCCGG GTCCTCAACG 3001 ACAGGAGCAC GATCATGCGC ACCCGTGGGG CCGCCATGCC GGCGATAATG GCCTGCTTCT 3061 CGCCGAAACG TTTGGTGGCG GGACCAGTGA CGAAGGCTTG AGCGAGGGCG TGCAAGATTC 3121 CGAATACCGC AAGCGACAGG CCGATCATCG TCGCGCTCCA GCGAAAGCGG TCCTCGCCGA 3181 AAATGACCCA GAGCGCTGCC GGCACCTGTC CTACGAGTTG CATGATAAAG AAGACAGTCA 3241 TAAGTGCGGC GACGATAGTC ATGCCCCGCG CCCACCGGAA GGAGCTGACT GGGTTGAAGG 3301 CTCTCAAGGG CATCGGTCGA GATCCCGGTG CCTAATGAGT GAGCTAACTT ACATTAATTG 3361 CGTTGCGCTC ACTGCCCGCT TTCCAGTCGG GAAACCTGTC GTGCCAGCTG CATTAATGAA 3421 TCGGCCAACG CGCGGGGAGA GGCGGTTTGC GTATTGGGCG CCAGGGTGGT TTTTCTTTTC 3481 ACCAGTGAGA CGGGCAACAG CTGATTGCCC TTCACCGCCT GGCCCTGAGA GAGTTGCAGC 3541 AAGCGGTCCA CGCTGGTTTG CCCCAGCAGG CGAAAATCCT GTTTGATGGT GGTTAACGGC 3601 GGGATATAAC ATGAGCTGTC TTCGGTATCG TCGTATCCCA CTACCGAGAT ATCCGCACCA 3661 ACGCGCAGCC CGGACTCGGT AATGGCGCGC ATTGCGCCCA GCGCCATCTG ATCGTTGGCA 3721 ACCAGCATCG CAGTGGGAAC GATGCCCTCA TTCAGCATTT GCATGGTTTG TTGAAAACCG 3781 GACATGGCAC TCCAGTCGCC TTCCCGTTCC GCTATCGGCT GAATTTGATT GCGAGTGAGA 3841 TATTTATGCC AGCCAGCCAG ACGCAGACGC GCCGAGACAG AACTTAATGG GCCCGCTAAC 3901 AGCGCGATTT GCTGGTGACC CAATGCGACC AGATGCTCCA CGCCCAGTCG CGTACCGTCT 3961 TCATGGGAGA AAATAATACT GTTGATGGGT GTCTGGTCAG AGACATCAAG AAATAACGCC 4021 GGAACATTAG TGCAGGCAGC TTCCACAGCA ATGGCATCCT GGTCATCCAG CGGATAGTTA 4081 ATGATCAGCC CACTGACGCG TTGCGCGAGA AGATTGTGCA CCGCCGCTTT ACAGGCTTCG 4141 ACGCCGCTTC GTTCTACCAT CGACACCACC ACGCTGGCAC CCAGTTGATC GGCGCGAGAT 4201 TTAATCGCCG CGACAATTTG CGACGGCGCG TGCAGGGCCA GACTGGAGGT GGCAACGCCA 4261 ATCAGCAACG ACTGTTTGCC CGCCAGTTGT TGTGCCACGC GGTTGGGAAT GTAATTCAGC 4321 TCCGCCATCG CCGCTTCCAC TTTTTCCCGC GTTTTCGCAG AAACGTGGCT GGCCTGGTTC 4381 ACCACGCGGG AAACGGTCTG ATAAGAGACA CCGGCATACT CTGCGACATC GTATAACGTT 4441 ACTGGTTTCA CATTCACCAC CCTGAATTGA CTCTCTTCCG GGCGCTATCA TGCCATACCG 4501 CGAAAGGTTT TGCGCCATTC GATGGTGTCC GGGATCTCGA CGCTCTCCCT TATGCGACTC 4561 CTGCATTAGG AAGCAGCCCA GTAGTAGGTT GAGGCCGTTG AGCACCGCCG CCGCAAGGAA 4621 TGGTGCATGC AAGGAGATGG CGCCCAACAG TCCCCCGGCC ACGGGGCCTG CCACCATACC 4681 CACGCCGAAA CAAGCGCTCA TGAGCCCGAA GTGGCGAGCC CGATCTTCCC CATCGGTGAT 4741 GTCGGCGATA TAGGCGCCAG CAACCGCACC TGTGGCGCCG GTGATGCCGG CCACGATGCG 4801 TCCGGCGTAG AGGATCGAGA TCTCGATCCC GCGAAATTAA TACGACTCAC TATAGGGGAA 4861 TTGTGAGCGG ATAACAATTC CCCTCTAGAA ATAATTTTGT TTAACTTTAA GAAGGAGATA 4921 TACCATGGGC AGCAGCCATC ATCATCATCA TCACAGCAGC GGCCTGGTGC CGCGCGGCAG 4981 CCATATGGCT AGCATGTCGG ACTCAGAAGT CAATCAAGAA GCTAAGCCAG AGGTCAAGCC 5041 AGAAGTCAAG CCTGAGACTC ACATCAATTT AAAGGTGTCC GATGGATCTT CAGAGATCTT 5101 CTTCAAGATC AAAAAGACCA CTCCTTTAAG AAGGCTGATG GAAGCGTTCG CTAAAAGACA 5161 GGGTAAGGAA ATGGACTCCT TAAGATTCTT GTACGACGGT ATTAGAATCC AAGCTGATCA 5221 GACCCCTGAA GATTTGGACA TGGAGGATAA CGATATTATT GAGGCTCACA GAGAACAGAT 5281 TGGTGGATCC GAATTCGAGC TCCGTCGACA AGCTTGCGGC CGCACTCGAG CACCACCACC 5341 ACCACCACTG AGATCCGGCT GCTAACAAAG CCCGAAAGGA AGCTGAGTTG GCTGCTGCCA 5401 CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTC
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