| 出品公司: | Novangen |
|---|---|
| 载体名称: | pET-52b, pET52b |
| 质粒类型: | 大肠杆菌表达载体；蛋白纯化 |
| 高拷贝/低拷贝: | 低拷贝 |
| 克隆方法: | 多克隆位点；限制性内切酶 |
| 启动子: | T7 |
| 载体大小: | 5227 bp |
| 5' 测序引物及序列: | T7 Fwd: 5'd[TAATACGACTCACTATAGGG]3' |
| 3' 测序引物及序列: | -- |
| 载体标签: | His Tag (C-末端)；Strep TagII (N-末端) |
| 载体抗性: | 氨苄青霉素（Ampicillin） |
| 备注: |
Strep TagII 下游含HRV 3C识别位点； His Tag 上游含Thrombin识别位点 |
| 产品目录号: | 71554-3 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 诱导型（IPTG) |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3)、C43(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
pET52b载体基本信息
pET52b载体图谱质粒图谱和多克隆位点信息
pET52b载体简介
pET-52b载体是带有N-Strep和HRV3C蛋白酶位点，可以在大肠杆菌中高表达目的蛋白。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。
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(Cat. No. 69337-3)的作用下终止蛋白翻译。 pET-52b载体上面含有一个HRV3C蛋白酶切位点，当将目的基因使用载体上面的PshAI限制性内切酶位点插入进去时，可以通过HRV3C蛋白酶将载体上的融合的所有氨基酸序列包括GST标签序列，全部切除掉。
pET52b载体序列
LOCUS pET-52b 5227 bp DNA circular 1-SEP-2005 DEFINITION pET-52b(+) or pET52b vector sequence. SOURCE ORGANISM COMMENT VNTNAME|pET-52b(+)| COMMENT VNTREPLTYPE|Plasmid COMMENT VNTEXTCHREPL|Bacteria FEATURES Location/Qualifiers misc_RNA complement(1656..1763) /vntifkey="53" /label=RNA\I misc_RNA 1648..2203 /vntifkey="53" /label=RNA\II CDS complement(2634..2825) /vntifkey="4" /label=ROP RBS 4944..4949 /vntifkey="32" /label=RBS-1 primer 4824..4839 /vntifkey="27" /label=pET\Upstream\Primer promoter 4870..4886 /vntifkey="30" /label=T7\promoter CDS complement(3397..4479) /vntifkey="4" /label=lac\I rep_origin 2203..2203 /vntifkey="33" /label=ColE1\pBR322\origin CDS complement(582..1442) /vntifkey="4" /label=bla\(amp)\sequence rep_origin 13..460 /vntifkey="33" /label=f1\origin CDS 4890..4911 /vntifkey="4" /label=lac\operator terminator 5155..5202 /vntifkey="43" /label=T7\Terminator -35_signal 1616..1621 /vntifkey="48" /label=-35\RNA\II -10_signal 1639..1644 /vntifkey="47" /label=-10\RNA\II misc_RNA 1652..2203 /vntifkey="53" /label=RNA\II -35_signal complement(1793..1798) /vntifkey="48" /label=-35\RNA\I -10_signal complement(1769..1774) /vntifkey="47" /label=-10\RNA\I misc_RNA complement(1626..1762) /vntifkey="53" /label=RNA\I terminator 1607..1625 /vntifkey="43" /label=RNA\I\Terminator protein_bind 4966..4989 /vntifkey="31" /label=Strep-Tag\II protein_bind 4996..5018 /vntifkey="31" /label=3c misc_feature 5065..5082 /vntifkey="21" /label=Thrombin\site misc_binding 5089..5119 /vntifkey="20" /label=10x\His-Tag primer 4990..5082 /vntifkey="27" /label=S-3cLIC\MCS primer complement(4996..5088) /vntifkey="27" /label=AS-3cLIC\MCS primer 4956..4989 /vntifkey="27" /label=S-NTerm\Strep primer complement(4960..4995) /vntifkey="27" /label=AS-NTerm\Strep BASE COUNT 1207 a 1401 c 1355 g 1264 t 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 caagctggag ccacccgcag 4981 ttcgaaaagg gtgcacttga agtcctcttt cagggacccg ggtaccagga tcctgtacaa 5041 gtcgacgcgg ccgcagagct cgctctggtg ccacgcggta gttccgctca tcaccaccat 5101 catcaccatc accaccacta attaacctag gctgctgcca ccgctgagca ataactagca 5161 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata 5221 tccggat //
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pET52b载体质粒应用举例
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