出品公司: | EMD Biosciences (Novagen) |
---|---|
別名: | pET45b, pET 45b |
質粒類型: | 大腸桿菌蛋白表達 |
表達水平: | 高 |
克隆方法: | 多克隆位點,限制性內切酶 |
載體大小: | 5260 bp |
5' 測序引物: | T7 |
5' 測序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 測序引物: | T7t |
3' 測序引物序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
載體標簽: | N-His,N-EK, C-S |
載體抗性: | Ampicillin |
備注: |
N-term HisTag with minimal extraneous sequences.
|
產品目錄號: | 71327-3 |
穩定性: | 瞬時表達 Transient |
組成型: | 組成型 Constitutive |
病毒/非病毒: | 非病毒 |
表達宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3)、C43(DE3) |
pET表達手冊: |
pET表達手冊英文版 pET表達手冊中文版 |
pET載體匯總: |
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 |
買家導航
pET-45b(+)載體 pET45b質粒基本信息
pET-45b(+)質粒圖譜 pET45b載體圖譜和pET-45b(+)載體序列 pET45b質粒序列多克隆位點信息
Feature Name | Start | End | |
---|---|---|---|
Ampicillin | 1442 | 582 | |
AmpR_promoter | 1513 | 1485 | |
pBR322_origin | 1600 | 2219 | |
pGEX_3_primer | 2618 | 2640 | |
ROP | 2825 | 2634 | |
lacI | 4488 | 3397 | |
tet (887 - 636) | 4831 | 4580 | |
pBRrevBam_primer | 4763 | 4782 | |
T7_promoter | 4870 | 4888 | |
lacO | 4888 | 4915 | |
T7_transl_en_RBS | 4933 | 4949 | |
EK | 4999 | 5013 | |
T7_Terminal_primer | 5192 | 5174 | |
T7_terminator | 5193 | 5228 |
ORF | Start | End | |
---|---|---|---|
ORF frame 3 | 1442 | 582 | |
ORF frame 2 | 4356 | 3397 | |
ORF frame 1 | 4729 | 4229 |
Enzyme Name | Cut |
---|---|
FspI | 877 |
HpaI | 3622 |
ApaI | 3921 |
BclI | 4110 |
ClaI | 4849 |
XbaI | 4916 |
NcoI | 4955 |
PmlI | 4980 |
KpnI | 4988 |
AgeI | 4987 |
BstBI | 4993 |
BamHI | 5019 |
SacI | 5036 |
AscI | 5047 |
PstI | 5057 |
SalI | 5059 |
HindIII | 5065 |
NotI | 5072 |
EagI | 5072 |
XhoI | 5080 |
PacI | 5155 |
pET-45b(+)質粒 pET45b載體簡介
pET-45b載體是帶有N-His蛋白標簽,EK蛋白酶位點和C-S標簽,可以在大腸桿菌中高表達目的蛋白。獨特的酶切位點在載體圖譜中被標注出來。需要注意的是,這些載體序列是依據載體pBR322的方向來進行編號的,所以T7標簽表達區域在載體圖譜上是反向的。
T7 RNA聚合酶啟動的克隆和表達區域在質粒圖譜中也被標注了出來。質粒的F1復制子是被定向的,所以在T7噬菌體聚合酶的作用下,包含有蛋白編碼序列的病毒 粒子能夠產生,并啟動蛋白表達,同時蛋白表達將被T7終止子序列(Cat. No. 69337-3)的作用下終止蛋白翻譯。 pET-45b載體上面含有一個EK蛋白酶切位點,當將目的基因使用載體上面的PshAI限制性內切酶位點插入進去時,可以通過EK蛋白酶將載體上的融合的所有氨基酸序列包括標簽序列,全部切除掉。
pET-45b(+)質粒序列 pET45b載體序列
LOCUS pET-45b(+) 5260 bp DNA Circular SYN DEFINITION pET-45b(+) ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. COMMENT This file is created by Vector NTI //gt12365.cn/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|gt12365.cn| COMMENT VNTOAUTHORAD1|Novagen Inc.| FEATURES Location/Qualifiers source 1..5260 /organism="pET-45b(+)" /mol_type="other DNA" gene complement(582..1442) /label="Ampicillin" /gene="Ampicillin" CDS complement(582..1442) /label="ORF frame 3" promoter complement(1485..1513) /label="AmpR_promoter" rep_origin 1600..2219 /label="pBR322_origin" misc_feature 2618..2640 /label="pGEX_3_primer" misc_feature complement(2634..2825) /label="ROP" misc_feature complement(3397..4488) /label="lacI" CDS complement(3397..4356) /label="ORF frame 2" CDS complement(4229..4729) /label="ORF frame 1" gene complement(4580..4831) /label="tet (887 - 636)" /gene="tet (887 - 636)" misc_feature 4763..4782 /label="pBRrevBam_primer" promoter 4870..4888 /label="T7_promoter" misc_feature 4888..4915 /label="lacO" misc_feature 4933..4949 /label="T7_transl_en_RBS" misc_feature 4999..5013 /label="EK" misc_feature complement(5174..5192) /label="T7_Terminal_primer" terminator 5193..5228 /label="T7_terminator" ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat 481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg 1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac 1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt 2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac 2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg 2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg 2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga 2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg 2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa 2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt 2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt 3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct 3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac 3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag 3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc 3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc 3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc 3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt 3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg 3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc 3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat 3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt 3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc 3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc 3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag 3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt 4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat 4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag 4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac 4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg 4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg 4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt 4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc 4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact 4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg 4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga 4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc 4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt 4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg 4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc 4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag 4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac 4981 gtgggtaccg gttcgaatga tgacgacgac aagagtccgg atcccaattg ggagctcgtg 5041 tacacggcgc gcctgcaggt cgacaagctt gcggccgcac tcgagtctgg taaagaaacc 5101 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc 5161 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc 5221 ttgaggggtt ttttgctgaa aggaggaact atatccggat //
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pET-45b(+)質粒 pET45b載體應用舉例
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