pET-47b - 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
XY0477	pET-47b	20μ濒	?请加蚕蚕询价

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20μ濒 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 promoter
复制子:	ColE1 ori，F1 ori
终止子:	T7 terminator
质粒分类:	大肠杆菌载体；笔贰罢系列表达质粒
质粒大小:	5203bp
质粒标签:	N-6×His, C-S, C-Thrombin
原核抗性:	卡那霉素碍补苍
克隆菌株:	大肠杆菌顿贬5α
培养条件:	37℃，有氧，尝叠
表达宿主:	BL21(DE3)
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:	T7:TAATACGACTCACTATAGGG
3'测序引物:	T7-ter:TGCTAGTTATTGCTCAGCGG

质粒介绍

pET47b载体携带有N端His纯化标签，紧跟着有一个HRV 3C蛋白酶切位点。这个HRV 3C蛋白酶切位点能够特异性的高效切割 “LEVLFQ↓GP”氨基酸序列，酶活力在低温下很强。pET-50b(+)载体上还含有一个可选择的C端Thrombin蛋白酶切位点和S标签序列。
pET-47b(+)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终止子序列的作用下终止蛋白翻译。

质粒图谱

质粒序列

LOCUS       Exported                5203 bp ds-DNA     circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
KEYWORDS    pET-47b
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 5203)
TITLE     Direct Submission
http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..5203
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      26..73
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(168..212)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /note="S-Tag"
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(213..230)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /note="thrombin site"
                     /translation="LVPRGS"
     CDS             complement(309..332)
                     /codon_start=1
                     /product="recognition and cleavage site for human
                     rhinovirus 3C and PreScission proteases"
                     /note="HRV 3C site"
                     /translation="LEVLFQGP"
     CDS             complement(342..359)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     protein_bind    410..434
                     /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(435..453)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        766..843
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             844..1926
                     /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             2498..2689
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     rep_origin      complement(3119..3707)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             3829..4644
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(4737..5192)
                     /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 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
      181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
      241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
      301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg gtggtgatgt
      361 gccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
      421 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga
      481 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
      541 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
      601 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
      661 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
      721 gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg
      781 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
      841 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
      901 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
      961 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     1021 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     1081 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     1141 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     1201 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     1261 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     1321 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     1381 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     1441 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     1501 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     1561 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     1621 tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc
     1681 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     1741 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     1801 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     1861 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1921 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc
     1981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactag
     2041 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
     2101 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
     2161 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
     2221 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
     2281 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
     2341 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
     2401 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     2461 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     2521 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     2581 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     2641 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2761 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     2821 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     2881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3541 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3781 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     3841 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
     3901 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
     3961 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     4021 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     4081 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
     4141 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     4201 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagtgat
     4261 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     4321 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     4381 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4441 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     4501 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     4561 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     4621 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
     4681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
     4741 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4801 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4861 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4921 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4981 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5041 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5101 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5161 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//