| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 别名: | pET31b, pet-31b(+) |
| 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: | 5742 bp |
| 5' 测序引物: | T7: 5'-TAATACGACTCACTATAGGG-3' |
| 3' 测序引物: | T7t: 5'-GCTAGTTATTGCTCAGCGG-3' |
| 载体标签: | C-His, N-KSI |
| 载体抗性: | 氨苄 |
| 备注: | 融合表达有N端酮类固醇异构酶，有助于提高蛋白及小肽的产量; AlwNI单克隆位点可以用来插入单向重复多肽序列。 |
| 产品目录号: | 69952-3 |
| 稳定性: | 瞬时表达 |
| 组成型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3)、C43(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
pET31b载体基本信息
pET31b载体图谱质粒图谱和多克隆位点信息
pET31b载体简介
pET-31b(+)载体(Cat. No. 69952-3)是设计用来高水平表达蛋白质的载体，载体融合表达有125个氨基酸的酮类固醇异构酶(ketosteroid isomerase protein) 。载体上面的AlwN I单克隆位点允许单向插入串联重复的通过蛋氨酸（methionine）分隔开的多肽序列。
pET31b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。
pET31b载体序列
LOCUS pET-31b(+) 5742 bp DNA Circular SYN
DEFINITION pET-31b(+)
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..5742
/organism="pET-31b(+)"
/mol_type="other DNA"
terminator complement(1..129)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
misc_feature complement(553..569)
/label="T7_transl_en_RBS"
misc_feature complement(587..614)
/label="lacO"
promoter complement(614..632)
/label="T7_promoter"
gene 664..927
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(716..735)
/label="pBRrevBam_primer"
CDS 769..1269
/label="ORF frame 1"
misc_feature 1010..2101
/label="lacI"
CDS 1142..2101
/label="ORF frame 2"
gene 2160..2435
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
misc_feature 2910..3101
/label="ROP"
misc_feature complement(3095..3117)
/label="pGEX_3_primer"
gene complement(4284..5144)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(4284..5144)
/label="ORF frame 2"
promoter complement(5186..5214)
/label="AmpR_promoter"
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagcagcatc tggcatgcgt
181 gaatattctt ctcgccaaac aaggcgcgga tgctcaccac cttgccggcg ccattgaagc
241 gaaagtgatc gatgggcgca actacggtct tgcggccctg atactcgaag ctgacggtga
301 aagcgaaggc cgcttcgttg gcgaccgcgc gtacctcctg cgtcagctcc accgccaaag
361 gcagtttgag cgagttggcg taaaactcac gaatcgcagc cgtaccggac ctgggctcgg
421 aacccacggg gtcttccacc gtggcgtcat cggcaaacag cgcgacgatg ccgtccagat
481 cgccggcatt gagcgcagcc acaaagcgct gtaccacggc ggtgatgtgt tctggggtat
541 gcatatgtat atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc
601 cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc
661 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
721 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
781 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
841 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
901 ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa
961 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
1021 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
1081 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
1141 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
1201 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
1261 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
1321 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
1381 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
1441 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
1501 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
1561 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
1621 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
1681 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
1741 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
1801 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
1861 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
1921 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
1981 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
2041 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
2101 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
2161 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
2221 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
2281 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
2341 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
2401 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cccacgggtg cgcatgatcg
2461 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
2521 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
2581 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
2641 cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa
2701 cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt ctctggtccc
2761 gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg gcatgttcat
2821 catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt acccccatga
2881 acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca
2941 tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg
3001 cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca
3061 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
3121 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
3181 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
3241 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatatgcgg
3301 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
3361 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
3421 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
3481 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
3541 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
3601 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
3661 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
3721 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
3781 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
3841 gagtccaacc cggtaagaca cgacttatcg ccactggcag ctggtaacag gattagcaga
3901 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3961 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4021 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
4081 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
4141 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
4201 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
4261 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
4321 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
4381 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
4441 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
4501 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
4561 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
4621 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
4681 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
4741 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
4801 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
4861 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
4921 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
4981 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
5041 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
5101 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
5161 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
5221 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt
5281 gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
5341 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
5401 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
5461 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
5521 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
5581 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
5641 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
5701 gccgcgctta atgcgccgct acagggcgcg tcccattcgc ca
//
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pET31b载体质粒应用举例
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