pET-30a - 91蜜桃视频

产物信息
基本信息
质粒介绍
质粒图谱
质粒序列

产物信息

产物货号	产物名称	产物规格	优惠价
XY0031	pET-30a	5μ驳	?请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产物的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子:	T7/lac
复制子:	ColE1 ori，F1 ori
终止子:	T7 terminator
质粒分类:	大肠系列质粒；大肠表达质粒；辫贰罢系列质粒
质粒大小:	5422bp
质粒标签:	N-6×His, N-S, N-Thrombin, N-EK, C-6×His
原核抗性:	卡那霉素碍补苍（50μ驳/尘濒）
克隆菌株:	顿贬5α等大肠杆菌
培养条件:	37℃，有氧，尝叠
表达宿主:	叠尝21(顿贰3)等大肠杆菌
培养条件:	37℃，有氧，尝叠
诱导方式:	滨笔罢骋或乳糖及其类似物
5'测序引物:	罢7（罢础础罢础颁骋础颁罢颁础颁罢础罢础骋骋骋）
3'测序引物:	罢7-迟别谤（罢骋颁罢础骋罢罢础罢罢骋颁罢颁础骋颁骋骋）

质粒介绍

pET-30a质粒是一种原核表达载体，C端含有一个6×His affinity标签，N端含有一个thrombin位点、6×His affinity标签、EK标签和S标签。该质粒含有多个常用的酶切位点，便于不同基因克隆。表达由宿主细胞提供的T7 RNA聚合酶诱导，目的基因被克隆到质粒载体上，受噬菌体强转录及翻译信号控制。
pET系统是有史以来在大肠杆菌中表达重组蛋白的功能*强大的系统，也是现今原核表达方面使用*广泛的系统。该系列质粒能很容易的通过降低诱导物的浓度来削弱蛋白表达。在非诱导条件下，可以使目的基因处于完全沉默状态而不转录。

The pET-30a-c(+) vectors carry an N-terminal His? Tag®/ thrombin/ S? Tag?/ enterokinase configuration plus an optional C-terminal His? Tag sequence. 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 .

质粒图谱

质粒序列

LOCUS       Exported                5422 bp ds-DNA     circular SYN 05-8-2015
DEFINITION synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 2
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 5422)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2015-8-5 from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5422
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          355..377
                     /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(219..233)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /note="enterokinase site"
                     /translation="DDDDK"
     CDS             complement(249..293)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(300..317)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(327..344)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     RBS             355..377
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    392..416
                     /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(417..435)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        748..825
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             826..1908
                     /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             2717..2908
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3010..3152
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3338..3926)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             4048..4863
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(4956..5411)
                     /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 gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgatatca gccatggcct tgtcgtcgtc gtcggtaccc
      241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatacca
      301 gaaccgcgtg gcaccagacc agaagaatga tgatgatgat ggtgcatatg tatatctcct
      361 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
      421 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
      481 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
      541 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
      601 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
      661 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
      721 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
      781 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
      841 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
      901 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
      961 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     1021 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     1081 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     1141 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     1201 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     1261 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     1321 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     1381 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     1441 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     1501 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     1561 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     1621 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     1681 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     1741 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     1801 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1861 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1921 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     1981 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2041 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     2101 cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg tattcggaat
     2161 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     2221 gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc tcctgtcgtt
     2281 gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg
     2341 cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat
     2401 ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat
     2461 tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt
     2521 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     2581 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     2641 tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc
     2701 ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat
     2761 cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc
     2821 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     2881 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2941 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3001 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     3061 tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt gaaataccgc
     3121 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     3181 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3241 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3301 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3361 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3421 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3481 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3541 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3601 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3661 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3721 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3781 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3841 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3901 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3961 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa actgtctgct
     4021 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctct
     4081 aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat
     4141 aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag
     4201 ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga
     4261 ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct
     4321 gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa
     4381 gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg
     4441 cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag
     4501 gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat
     4561 ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat
     4621 tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta
     4681 ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc
     4741 ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat
     4801 ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc
     4861 taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4921 ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta atattttgtt
     4981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     5041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     5101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     5161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     5221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     5281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     5341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     5401 acagggcgcg tcccattcgc ca
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