出品公司: | EMD Biosciences(Novagen) |
---|---|
别名: | pET33b, pET 33b |
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: | 5383 bp |
5' 测序引物: | T7 |
5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
载体标签: | His (中间和C端), N-Thrombin |
载体抗性: | Kanamycin |
备注: |
Production of target proteins suitable for site-specific 32P-labeling; Nterm thrombin cleavage site; Nterm enterokinase cleavage site
|
产品目录号: | 69054-3 |
稳定性: | 瞬时表达 Transient |
组成型: | 组成型 Constitutive |
病毒/非病毒: | 非病毒 |
表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(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
pET-33b(+)载体 pET33b质粒基本信息
pET-33b(+)质粒图谱 pET33b载体图谱和pET-33b(+)载体序列 pET33b质粒序列多克隆位点信息
Feature Name | Start | End | |
---|---|---|---|
T7_terminator | 129 | 1 | |
T7_Terminal_primer | 69 | 87 | |
T7_leader | 238 | 206 | |
Xpress_fwd_primer | 239 | 221 | |
T7_transl_en_RBS | 337 | 321 | |
lacO | 382 | 355 | |
T7_promoter | 400 | 382 | |
tet (300 - 563) | 432 | 695 | |
pBRrevBam_primer | 503 | 484 | |
lacI | 778 | 1869 | |
tet (576 - 851) | 1928 | 2203 | |
ROP | 2678 | 2869 | |
pGEX_3_primer | 2885 | 2863 | |
pBR322_origin | 3903 | 3284 | |
KanR2 | 4009 | 4824 |
ORF | Start | End | |
---|---|---|---|
ORF frame 3 | 537 | 1037 | |
ORF frame 1 | 910 | 1869 | |
ORF frame 1 | 4009 | 4824 |
Enzyme Name | Cut |
---|---|
XhoI | 158 |
NotI | 166 |
EagI | 166 |
HindIII | 173 |
SalI | 179 |
SacI | 190 |
EcoRI | 192 |
BamHI | 198 |
NheI | 230 |
NdeI | 237 |
NcoI | 310 |
XbaI | 349 |
BglII | 415 |
BclI | 1151 |
ApaI | 1348 |
EcoRV | 1587 |
HpaI | 1643 |
FspI | 2219 |
NruI | 4097 |
ClaI | 4131 |
XmaI | 4312 |
SmaI | 4314 |
pET-33b(+)质粒 pET33b载体简介
pET-33b(+)载体(目录号69054-3)来源于pET-28b(+),携带一段编码蛋白激酶A(PKA)位点RRASV的15bp序列,位于Thrombin和Nde I位点(1)之间。pET-33b(+)中表达的蛋白质可以通过镍纯化柱(通过N-或C-末端His•Tag序列)轻松纯化,并用32P-或33P-gATP以及心肌中的cAMP依赖性蛋白激酶的催化亚单位进行有效标记。标记蛋白质可以作为蛋白质相互作用研究中的直接探针。独特的位点显示在载体图谱上。请注意,序列是按照pBR322正向方向编号的,因此T7表达区域在图谱上是反向的。T7 RNA聚合酶转录的编码链的克隆/表达区域如下所示。f1起源是定向的,因此辅助噬菌体感染将产生包含与编码链对应的单链DNA的病毒粒子。因此,应使用T7终止子引物(目录号69337-3)进行单链测序。
pET-33b(+)质粒序列 pET33b载体序列
LOCUS pET-33b(+) 5383 bp DNA Circular SYN DEFINITION pET-33b(+) 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..5383 /organism="pET-33b(+)" /mol_type="other DNA" terminator complement(1..129) /label="T7_terminator" misc_feature 69..87 /label="T7_Terminal_primer" misc_feature complement(206..238) /label="T7_leader" misc_feature complement(221..239) /label="Xpress_fwd_primer" misc_feature complement(321..337) /label="T7_transl_en_RBS" misc_feature complement(355..382) /label="lacO" promoter complement(382..400) /label="T7_promoter" gene 432..695 /label="tet (300 - 563)" /gene="tet (300 - 563)" misc_feature complement(484..503) /label="pBRrevBam_primer" CDS 537..1037 /label="ORF frame 3" /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV ANCRGD*" misc_feature 778..1869 /label="lacI" /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV ANCRGD*" CDS 910..1869 /label="ORF frame 1" /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*" gene 1928..2203 /label="tet (576 - 851)" /gene="tet (576 - 851)" /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*" misc_feature 2678..2869 /label="ROP" /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*" misc_feature complement(2863..2885) /label="pGEX_3_primer" /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*" rep_origin complement(3284..3903) /label="pBR322_origin" /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*" gene 4009..4824 /label="KanR2" /gene="KanR2" /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*" CDS 4009..4824 /label="ORF frame 1" /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 gagtgcggcc gcaagcttgt 181 cgacggagct cgaattcgga tcccgaccca tttgctgtcc accagtcatg ctagccatat 241 gaacagatgc acgacggctg ccgcgcggca ccaggccgct gctgtgatga tgatgatgat 301 ggctgctgcc catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa 361 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct 421 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg 481 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga 541 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca 601 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg 661 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa 721 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg 781 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag 841 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg 901 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc 961 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa 1021 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg 1081 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc 1141 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct 1201 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt 1261 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt 1321 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg 1381 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc 1441 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt 1501 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc 1561 gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac 1621 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa 1681 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg 1741 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct 1801 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc 1861 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg 1921 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac 1981 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc 2041 agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct 2101 gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc 2161 caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccc cacgggtgcg 2221 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta 2281 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc 2341 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg 2401 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc 2461 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct 2521 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc 2581 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac 2641 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc 2701 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga 2761 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct 2821 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct 2881 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 2941 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag 3001 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 3061 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt 3121 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 3181 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 3241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 3301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 3361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 3421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 3481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 3541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 3601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 3661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 3721 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 3781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 3841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 3901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 3961 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt 4021 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat 4081 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat 4141 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat 4201 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc 4261 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa 4321 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg 4381 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat 4441 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt 4501 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa 4561 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt 4621 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac 4681 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag 4741 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat 4801 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat 4861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat 4921 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 4981 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 5041 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 5101 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 5161 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 5221 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 5281 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 5341 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca //
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pET-33b(+)质粒 pET33b载体应用举例
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