| 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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