| 出品公司: | EMD Biosciences (Novagen) |
|---|---|
| 别名: | pET24b, pet 24b |
| 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: | 5309 bp |
| 5' 测序引物: | T7 |
| 5' 测序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 载体标签: | N-T7, C-His |
| 载体抗性: | Kanamycin |
| 备注: |
Same as pET21a,b,c,d(+) but kanamycin resistance; a,b,c,d vary by MCS; The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand.
|
| 产品目录号: | 69750-3 |
| 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-24b(+)载体 pET24b质粒基本信息
pET-24b(+)质粒图谱 pET24b载体图谱和pET-24b(+)载体序列 pET24b质粒序列多克隆位点信息
The maps for pET-24b(+), pET-24c(+) and pET-24d(+) are the same as pET-24a(+) (shown) with the following exceptions: pET-24b(+) is a 5309bp plasmid; subtract 1bp from each site beyond BamH I at 198. pET-24c(+) is a 5308bp plasmid; subtract 2bp from each site beyond BamH I at 198. pET-24d(+) is a 5307bp plasmid; the BamH I site is in the same reading frame as in pET- 24c(+). An Nco I site is substituted for the Nde I site with a net 1bp deletion at position 238 of pET-24c(+). As a result, Nco I cuts pET24d(+) at 234, and Nhe I cuts at 229. For the rest of the sites, subtract 3bp from each site beyond position 239 in pET-24a(+). Nde I does not cut pET-24d(+). Note also that Sty I is not unique in pET-24d(+).
| 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 | 263 | 247 | |
| lacO | 308 | 281 | |
| T7_promoter | 326 | 308 | |
| tet (300 - 563) | 358 | 621 | |
| pBRrevBam_primer | 429 | 410 | |
| lacI | 704 | 1795 | |
| tet (576 - 851) | 1854 | 2129 | |
| ROP | 2604 | 2795 | |
| pGEX_3_primer | 2811 | 2789 | |
| pBR322_origin | 3829 | 3210 | |
| KanR2 | 3935 | 4750 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 1 | 463 | 963 | |
| ORF frame 2 | 836 | 1795 | |
| ORF frame 2 | 3935 | 4750 |
| Enzyme Name | Cut |
|---|---|
| XhoI | 158 |
| NotI | 166 |
| EagI | 166 |
| HindIII | 173 |
| SalI | 179 |
| SacI | 190 |
| EcoRI | 192 |
| BamHI | 198 |
| NheI | 230 |
| NdeI | 237 |
| XbaI | 275 |
| BglII | 341 |
| BclI | 1077 |
| ApaI | 1274 |
| EcoRV | 1513 |
| HpaI | 1569 |
| FspI | 2145 |
| NruI | 4023 |
| ClaI | 4057 |
| XmaI | 4238 |
| SmaI | 4240 |
pET-24b(+)质粒 pET24b载体简介
pET-24a-d系列载体带有N-T7标签和C-His标签。pET-24a-d系列载体与pET-21a-d系列载体相比，只有抗性不同。独特的酶切位点在载体图谱中被标注出来。需要注意的是，这些载体序列是依据载体pBR322的方向来进行编号的，所以T7标签表达区域在载体图谱上是反向的。
pET-24b(+)质粒序列 pET24b载体序列
LOCUS pET-24b (+) 5309 bp DNA circular SYN
DEFINITION pET-24b (+)
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..5309
/organism="pET-24 b (+)"
/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(247..263)
/label="T7_transl_en_RBS"
misc_feature complement(281..308)
/label="lacO"
promoter complement(308..326)
/label="T7_promoter"
gene 358..621
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(410..429)
/label="pBRrevBam_primer"
CDS 463..963
/label="ORF frame 1"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
misc_feature 704..1795
/label="lacI"
/translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
ANCRGD*"
CDS 836..1795
/label="ORF frame 2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 1854..2129
/label="tet (576 - 851)"
/gene="tet (576 - 851)"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature 2604..2795
/label="ROP"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
misc_feature complement(2789..2811)
/label="pGEX_3_primer"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
rep_origin complement(3210..3829)
/label="pBR322_origin"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
gene 3935..4750
/label="KanR2"
/gene="KanR2"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
CDS 3935..4750
/label="ORF frame 2"
/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 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
301 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
361 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
421 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
481 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
541 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
601 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
661 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
721 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
781 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
841 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
901 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
961 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
1021 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
1081 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
1141 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
1201 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
1261 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
1321 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
1381 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
1441 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
1501 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
1561 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
1621 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
1681 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
1741 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
1801 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
1861 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
1921 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
1981 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
2041 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
2101 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
2161 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
2221 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
2281 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
2341 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
2401 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
2461 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
2521 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
2581 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
2641 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
2701 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
2761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
2821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
2881 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
2941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
3001 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
3061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
3121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3661 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
3901 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
3961 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
4021 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
4081 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
4141 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
4201 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
4261 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
4321 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
4381 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
4441 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
4501 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
4561 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
4621 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
4681 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
4741 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
4801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
4861 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
4921 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
4981 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
5041 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
5101 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
5161 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
5221 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
5281 cgccgctaca gggcgcgtcc cattcgcca
//
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pET-24b(+)质粒 pET24b载体应用举例
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