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pET系列质粒

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:


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 CCCTGATAGA
121      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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