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產(chǎn)品名稱：pET-31b
產(chǎn)品型號(hào)：
產(chǎn)品展商：XYbscience
產(chǎn)品文檔：無(wú)相關(guān)文檔

簡(jiǎn)單介紹

pET-31b是一種原核表達(dá)載體，C端含有一個(gè)6×His標(biāo)簽，表達(dá)由宿主細(xì)胞提供的T7 RNA聚合酶誘導(dǎo)，目的基因被克隆到質(zhì)粒載體上，受噬菌體強(qiáng)轉(zhuǎn)錄及翻譯信號(hào)控制。 pET系統(tǒng)是有史以來(lái)在大腸桿菌中表達(dá)重組蛋白的功能*強(qiáng)???的系統(tǒng)，也是現(xiàn)今原核表達(dá)方面使用*廣泛的系統(tǒng)。pET-31b該系列質(zhì)粒能很容易的通過(guò)降低誘導(dǎo)物的濃度來(lái)削弱蛋白表達(dá)。在非誘導(dǎo)條件下，可以使目的基因處于完全沉默狀態(tài)而不轉(zhuǎn)錄。

產(chǎn)品描述

pET-31b

產(chǎn)品信息
基本信息
質(zhì)粒簡(jiǎn)介
質(zhì)粒圖譜
質(zhì)粒序列

產(chǎn)品信息

產(chǎn)品貨號(hào)	產(chǎn)品名稱	產(chǎn)品規(guī)格	優(yōu)惠價(jià)
XY0032	pET-31b	5μg	￥請(qǐng)?jiān)儍r(jià)

使用說(shuō)明

信裕質(zhì)粒平臺(tái)的各批次質(zhì)粒菌株發(fā)貨前均經(jīng)過(guò)嚴(yán)格的多重驗(yàn)證，如存在質(zhì)量問(wèn)題，請(qǐng)?jiān)谑盏疆a(chǎn)品的三個(gè)月內(nèi)通知我司。收到質(zhì)粒后請(qǐng)短暫離心，加入20μl ddH2O溶解質(zhì)粒，取2μl轉(zhuǎn)化至對(duì)應(yīng)感受態(tài)中，挑取單克隆重新提取質(zhì)粒后使用。如需獲取其他詳細(xì)信息請(qǐng)登錄我司官網(wǎng)查詢。

基本信息

啟動(dòng)子:	T7/lac
復(fù)制子:	ColE1 ori，F(xiàn)1 ori
終止子:	T7 terminator
質(zhì)粒分類:	大腸系列質(zhì)粒；大腸表達(dá)質(zhì)粒；pET系列質(zhì)粒
質(zhì)粒大小:	5742bp
質(zhì)粒標(biāo)簽:	C-6×His
原核抗性:	氨芐青霉素Amp（100μg/ml）
克隆菌株:	DH5α等大腸桿菌
培養(yǎng)條件:	37℃，有氧，LB
表達(dá)宿主:	BL21(DE3)等大腸桿菌
培養(yǎng)條件:	37℃，有氧，LB
誘導(dǎo)方式:	IPTG或乳糖及其類似物
5'測(cè)序引物:	T7（TAATACGACTCACTATAGGG）
3'測(cè)序引物:	T7-ter（TGCTAGTTATTGCTCAGCGG）
備注:	表達(dá)酮固醇異構(gòu)酶

質(zhì)粒簡(jiǎn)介

pET-31b質(zhì)粒是一種原核表達(dá)載體，C端含有一個(gè)6×His標(biāo)簽，表達(dá)由宿主細(xì)胞提供的T7 RNA聚合酶誘導(dǎo)，目的基因被克隆到質(zhì)粒載體上，受噬菌體強(qiáng)轉(zhuǎn)錄及翻譯信號(hào)控制。
pET系統(tǒng)是有史以來(lái)在大腸桿菌中表達(dá)重組蛋白的功能*強(qiáng)大的系統(tǒng)，也是現(xiàn)今原核表達(dá)方面使用*廣泛的系統(tǒng)。該系列質(zhì)粒能很容易的通過(guò)降低誘導(dǎo)物的濃度來(lái)削弱蛋白表達(dá)。在非誘導(dǎo)條件下，可以使目的基因處于完全沉默狀態(tài)而不轉(zhuǎn)錄。
The pET-31b(+) vector is designed for cloning and high-level expression of peptide sequences fused with the 125aa ketosteroid isomerase protein (1). The unique AlwN I cloning site allows the unidirectional insertion of of tandemly repeated peptide coding regions separated by methionine codons. 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 .

質(zhì)粒圖譜

質(zhì)粒序列

LOCUS       Exported                5742 bp ds-DNA     circular SYN 05-8-2015
DEFINITION synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 7
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 5742)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2015-8-5 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5742
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          552..574
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     CDS             complement(170..544)
                     /codon_start=1
                     /product="ketosteroid isomerase"
                     /note="KSI"
                     /translation="MHTPEHITAVVQRFVAALNAGDLDGIVALFADDATVEDPVGSEPR
                     SGTAAIREFYANSLKLPLAVELTQEVRAVANEAAFAFTVSFEYQGRKTVVAPIDHFRFN
                     GAGKVVSIRALFGEKNIHACQ"
     RBS             552..574
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    589..613
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(614..632)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        941..1018
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             1019..2101
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             2910..3101
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3203..3345
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(4284..5144)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5145..5249)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      complement(5276..5731)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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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產(chǎn)品目錄

產(chǎn)品資料

pET-31b

pET-31b

產(chǎn)品信息

使用說(shuō)明

基本信息

質(zhì)粒簡(jiǎn)介

質(zhì)粒圖譜

質(zhì)粒序列