pET-27b(+)大肠杆菌表达载体说明

pET-27b(+)

编号 载体名称

北京华越洋生物VECT4180 pET-­‐27b(+)

pET27b载体基本信息

别名: pET27b, p et 27b

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: 5414 b p

5' 测序引物: T7

5' 测序引物序列: 5'-­‐TAATACGACTCACTATAGGG-­‐3'

载体标签: N-­‐pelB, C-­‐His, C-­‐HSV

载体抗性: Kanamycin

备注: PelB sequence for periplasmic localization; same as pET25b but

kanR

稳定性: 瞬时表达 Transient

组成型: 组成型 Constitutive

病毒/非病毒: 非病毒

pET27b载体质粒图谱和多克隆位点信息

pET27b载体简介

The pET-­‐27b(+) vector (Cat. No. 69863-­‐3) carries an N-­‐terminal pelB signal sequence for potential p eriplasmic l ocalization, p lus o ptional C-­‐terminal H SV•Tag® a nd H is•Tag® s equences. This vector differs from pET-­‐25b(+) only by its selectable marker (kanamycin vs. ampicillin resistance).Unique s ites a re s hown o n t he c ircle m ap. N ote t hat t he s equence i s n umbered b y t he pBR322 convention, so the T7 expression region is reversed on the circular 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 T7 terminator primer (Cat. No. 69337-­‐3).

pET27b载体序列

ORIGIN

1 ATCCGGATAT AGTTCCTCCT TTCAGCAAAA AACCCCTCAA GACCCGTTTA GAGGCCCCAA

61 GGGGTTATGC TAGTTATTGC TCAGCGGTGG CAGCAGCCAA CTCAGCTTCC TTTCGGGCTT

121 TGTTAGCAGC CGGATCTCAG TGGTGGTGGT GGTGGTGCTC GACATCCTCG GGGTCTTCCG

181 GGGCGAGTTC TGGCTGGCTA GCCCGTTTGA TCTCGAGTGC GGCCGCAAGC TTGTCGACGG

241 AGCTCGAATT CGGATCCGAA TTAATTCCGA TATCCATGGC CATCGCCGGC TGGGCAGCGA 301 GGAGCAGCAG ACCAGCAGCA GCGGTCGGCA GCAGGTATTT CATATGTATA TCTCCTTCTT 361 AAAGTTAAAC AAAATTATTT CTAGAGGGGA ATTGTTATCC GCTCACAATT CCCCTATAGT 421 GAGTCGTATT AATTTCGCGG GATCGAGATC TCGATCCTCT ACGCCGGACG CATCGTGGCC 481 GGCATCACCG GCGCCACAGG TGCGGTTGCT GGCGCCTATA TCGCCGACAT CACCGATGGG 541 GAAGATCGGG CTCGCCACTT CGGGCTCATG AGCGCTTGTT TCGGCGTGGG TATGGTGGCA 601 GGCCCCGTGG CCGGGGGACT GTTGGGCGCC ATCTCCTTGC ATGCACCATT CCTTGCGGCG 661 GCGGTGCTCA ACGGCCTCAA CCTACTACTG GGCTGCTTCC TAATGCAGGA GTCGCATAAG 721 GGAGAGCGTC GAGATCCCGG ACACCATCGA ATGGCGCAAA ACCTTTCGCG GTATGGCATG 781 ATAGCGCCCG GAAGAGAGTC AATTCAGGGT GGTGAATGTG AAACCAGTAA CGTTATACGA 841 TGTCGCAGAG TATGCCGGTG TCTCTTATCA GACCGTTTCC CGCGTGGTGA ACCAGGCCAG 901 CCACGTTTCT GCGAAAACGC GGGAAAAAGT GGAAGCGGCG ATGGCGGAGC TGAATTACAT 961 TCCCAACCGC GTGGCACAAC AACTGGCGGG CAAACAGTCG TTGCTGATTG GCGTTGCCAC 1021 CTCCAGTCTG GCCCTGCACG CGCCGTCGCA AATTGTCGCG GCGATTAAAT CTCGCGCCGA 1081 TCAACTGGGT GCCAGCGTGG TGGTGTCGAT GGTAGAACGA AGCGGCGTCG AAGCCTGTAA 1141 AGCGGCGGTG CACAATCTTC TCGCGCAACG CGTCAGTGGG CTGATCATTA ACTATCCGCT 1201 GGATGACCAG GATGCCATTG CTGTGGAAGC TGCCTGCACT AATGTTCCGG CGTTATTTCT 1261 TGATGTCTCT GACCAGACAC CCATCAACAG TATTATTTTC TCCCATGAAG ACGGTACGCG 1321 ACTGGGCGTG GAGCATCTGG TCGCATTGGG TCACCAGCAA ATCGCGCTGT TAGCGGGCCC 1381 ATTAAGTTCT GTCTCGGCGC GTCTGCGTCT GGCTGGCTGG CATAAATATC TCACTCGCAA 1441 TCAAATTCAG CCGATAGCGG AACGGGAAGG CGACTGGAGT GCCATGTCCG GTTTTCAACA 1501 AACCATGCAA ATGCTGAATG AGGGCATCGT TCCCACTGCG ATGCTGGTTG CCAACGATCA 1561 GATGGCGCTG GGCGCAATGC GCGCCATTAC CGAGTCCGGG CTGCGCGTTG GTGCGGATAT 1621 CTCGGTAGTG GGATACGACG ATACCGAAGA CAGCTCATGT TATATCCCGC CGTTAACCAC 1681 CATCAAACAG GATTTTCGCC TGCTGGGGCA AACCAGCGTG GACCGCTTGC TGCAACTCTC 1741 TCAGGGCCAG GCGGTGAAGG GCAATCAGCT GTTGCCCGTC TCACTGGTGA AAAGAAAAAC 1801 CACCCTGGCG CCCAATACGC AAACCGCCTC TCCCCGCGCG TTGGCCGATT CATTAATGCA 1861 GCTGGCACGA CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGCAACGCA ATTAATGTAA 1921 GTTAGCTCAC TCATTAGGCA CCGGGATCTC GACCGATGCC CTTGAGAGCC TTCAACCCAG 1981 TCAGCTCCTT CCGGTGGGCG CGGGGCATGA CTATCGTCGC CGCACTTATG ACTGTCTTCT 2041 TTATCATGCA ACTCGTAGGA CAGGTGCCGG CAGCGCTCTG GGTCATTTTC GGCGAGGACC 2101 GCTTTCGCTG GAGCGCGACG ATGATCGGCC TGTCGCTTGC GGTATTCGGA ATCTTGCACG 2161 CCCTCGCTCA AGCCTTCGTC ACTGGTCCCG CCACCAAACG TTTCGGCGAG AAGCAGGCCA 2221 TTATCGCCGG CATGGCGGCC CCACGGGTGC GCATGATCGT GCTCCTGTCG TTGAGGACCC 2281 GGCTAGGCTG GCGGGGTTGC CTTACTGGTT AGCAGAATGA ATCACCGATA CGCGAGCGAA 2341 CGTGAAGCGA CTGCTGCTGC AAAACGTCTG CGACCTGAGC AACAACATGA ATGGTCTTCG 2401 GTTTCCGTGT TTCGTAAAGT CTGGAAACGC GGAAGTCAGC GCCCTGCACC ATTATGTTCC 2461 GGATCTGCAT CGCAGGATGC TGCTGGCTAC CCTGTGGAAC ACCTACATCT GTATTAACGA 2521 AGCGCTGGCA TTGACCCTGA GTGATTTTTC TCTGGTCCCG CCGCATCCAT ACCGCCAGTT 2581 GTTTACCCTC ACAACGTTCC AGTAACCGGG CATGTTCATC ATCAGTAACC CGTATCGTGA 2641 GCATCCTCTC TCGTTTCATC GGTATCATTA CCCCCATGAA CAGAAATCCC CCTTACACGG 2701 AGGCATCAGT GACCAAACAG GAAAAAACCG CCCTTAACAT GGCCCGCTTT ATCAGAAGCC 2761 AGACATTAAC GCTTCTGGAG AAACTCAACG AGCTGGACGC GGATGAACAG GCAGACATCT 2821 GTGAATCGCT TCACGACCAC GCTGATGAGC TTTACCGCAG CTGCCTCGCG CGTTTCGGTG