| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 别名: | pET27b, pet 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 bp |
| 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 |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、BL21(DE3)pLysS |
| 备注: |
含有pelB信号肽,能够将目的蛋白定位于细胞周质腔,除抗性不同外,其它方面均与pET-25b相同。 |
| 产品目录号: | ZY69863-3 |
| 稳定性: | 瞬时表达 Transient |
| 组成型: | 组成型 Constitutive |
| 病毒/非病毒: | 非病毒 |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
pET-27b(+)
- 库存:
20
- 供应商:
泽叶生物
pET27b载体基本信息
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 periplasmic localization, plus optional C-terminal HSV•Tag® and His•Tag® sequences. This vector differs from pET-25b(+) only by its selectable marker (kanamycin vs. ampicillin resistance).Unique sites are shown on the circle map. Note that the sequence is numbered by the 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
DEFINITION pET-27b(+)
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..5414
/organism="pET-27b(+)"
/mol_type="other DNA"
terminator complement(1..129)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
misc_feature complement(352..368)
/label="T7_transl_en_RBS"
misc_feature complement(386..413)
/label="lacO"
promoter complement(413..431)
/label="T7_promoter"
gene 463..726
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(515..534)
/label="pBRrevBam_primer"
CDS 568..1068
/label="ORF frame 1"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
misc_feature 809..1900
/label="lacI"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
CDS 941..1900
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 1959..2234
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 2709..2900
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(2894..2916)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(3315..3934)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 4040..4855
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 4040..4855
/label="ORF frame 2"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
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
2881 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
3001 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
3061 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
3121 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
3481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
3961 aacgaaaact cacgttaagg gattttggtc atgaacaata aaactgtctg cttacataaa
4021 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
4081 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
4141 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
4201 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
4261 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
4321 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
4381 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
4441 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
4501 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
4561 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
4621 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
4681 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
4741 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
4801 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
4861 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
4921 gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg
4981 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
5041 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
5101 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
5161 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
5221 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
5281 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
5341 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
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文献和实验表达载体的构建是生物技术和所需蛋白质产生的基本工具,本文总结了pET-32α(+)载体技术的构建,该技术通常用于研究实验室。该方法包括获得用于构建载体的外源DNA片段,将DNA片段亚克隆到pET-32α(+)表达载体中,在大肠杆菌 BL21(DE3)中进行蛋白质表达以及在大肠杆菌中在天然条件下进行蛋白质纯化。裂解液。介绍作为分子生物学实验室中非常有用的实践,重组DNA技术(或基因克隆)是指将DNA片段从一种生物体转移到自我复制的遗传元件如表达载体。然后插入的DNA可以在外源宿主细胞中繁殖
相关专题 1)噬菌体DE3溶源化的菌株如BL21(DE3)是最常用的表达菌株,噬菌体DE3是λ噬菌体的衍生株,构建好的表达载体 可以直接转入表达菌株中,诱导蛋白的表达方式与lac启动子一样都是IPTG诱导。 2)表达菌株BL21(DE3)上带有lacI基因,T7 RNA 聚合酶编码基因以及lacUV5启动子,lacUV5启动子可以启动T7 RNA 聚合酶的表达; 3)表达质粒 如pET-28质粒带有T7启动子和编码阻遏
Expression System、HisPatch ThioFusion System、pBAD Inducible Bacterial System、pL System和T7-based Systems。这四个系列的载体可谓把原核表达的发展史中出现过的几种启动子都囊括进去了,包括T7、pL、pBAD和trc启动子,同时几种经典的调控方式也都出现了,包括有乳糖操纵子、色氨酸操纵子和阿拉伯糖操纵子。 冠军系列 Champion™ pET Expression System之所以取
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