出品公司: | EMD Biosciences (Novagen) |
---|---|
别名: | pET50b, pET 50b |
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: | 6733 bp |
5' 测序引物及序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 测序引物及序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
载体标签: | N-Nus, N-His, N-HRV 3C, C-S, C-Thrombin |
载体抗性: | 卡那 |
备注: |
和pET47系列载体一样,但是多出了一个N端Nus标签;载体含有HRV 3C蛋白酶切位点,可以在低温下直接将多余的融合表达标签切除。
|
产品目录号: | 71464-3 |
稳定性: | 瞬时表达 |
组成型: | 组成型 |
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-50b(+)载体 pET50b质粒基本信息
pET-50b(+)质粒图谱 pET50b载体图谱和pET-50b(+)载体序列 pET50b质粒序列多克隆位点信息
pET-50b(+)质粒 pET50b载体简介
pET50b载体携带有N端的Nus标签和His纯化标签,紧跟着这些标签的有一个HRV3C蛋白酶切位点。这个HRV3C蛋白酶切位点能够特异性的高效切割 “LEVLFQ↓GP”氨基酸序列,酶活力在低温下很强。pET-50b(+)载体上还含有一个可选择的C端Thrombin蛋白酶切位点和S标签序列。
载体的单一的多克隆位点见上面的环状质粒图谱。注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面是反向的。
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-50b(+)质粒序列 pET50b载体序列
LOCUS pET-50b(+) 6733 bp DNA circular 11-SEP-2008 DEFINITION pET-50b(+). SOURCE ORGANISM COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTOAUTHORAD1|Novagen Inc.| COMMENT VNTREPLTYPE|Plasmid COMMENT VNTEXTCHREPL|Bacteria FEATURES Location/Qualifiers rep_origin complement(6274..6721) /vntifkey="33" /label=f1\ori misc_RNA complement(4650..5205) /vntifkey="53" /label=RNA\II rep_origin complement(4650..4650) /vntifkey="33" /label=ColE1,\pBR\ori misc_RNA 5090..5197 /vntifkey="53" /label=RNA\I CDS 4028..4219 /vntifkey="4" /label=ROP\gene CDS 2374..3456 /vntifkey="4" /label=lacI /note="lac repressor" promoter complement(1967..1983) /vntifkey="30" /label=T7\promoter protein_bind complement(1942..1962) /vntifkey="31" /label=lacO protein_bind complement(342..359) /vntifkey="31" /label=His-Tag terminator complement(26..73) /vntifkey="43" /label=T7\terminator CDS 5359..6174 /vntifkey="4" /label=Kan misc_feature complement(213..230) /vntifkey="21" /label=Thrombin\site protein_bind complement(168..212) /vntifkey="31" /label=S-Tag misc_binding complement(1866..1883) /vntifkey="20" /label=His-Tag CDS complement(375..1859) /vntifkey="4" /label=Nus-Tag protein_bind complement(310..332) /vntifkey="31" /label=3c primer complement(300..336) /vntifkey="27" /label=S-SacII/Bam\+SanDI\-dcm BASE COUNT 1584 a 1768 c 1759 g 1622 t ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc 181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct 241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc 301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg atggtgatga 361 ccagaaccac tagtcgcttc gtcaccgaac cagcaaatat tacgggcagc cataatcagt 421 gctccggctt tttcgtcggt caacccttcg atatcagcca gatcatcaat gccctgttcg 481 gcgagatctt ccagcgtaca aacgccacgg gcggccagtt tgaatgccaa atcacgatct 541 accccttcaa ggttcagcag atcgtcagcc ggtttgttat caccgaggct ttcttcctgg 601 gcctgtgcaa tggtggccag tgcattttta gcacgctcgc gcagtgcttc aacggtcggc 661 tcatcaaggc cttcgatttc caacagctct ttcatcggca cataggccaa ttcttccagc 721 gtcgagaagc cttcttctac cagaacagtc gcgaagtctt cgtcgatgtc gagatatttg 781 gtgaaggtgt cgatcgctgc gtgcgcttcc gcctgatgct tagcttgcag gtcgtcaacg 841 gtcatcacgt tgagttccca accgctcagt tgcgaagcca gacgcacgtt ctgaccgtta 901 cggccaatcg cctgcgccag attaccggct tcaacggcga tgtccatggt gtgtttatct 961 tcatccacca cgatagaagc aacgtctgcc ggtgccattg cgttaatcac gaactgcgcc 1021 gggttatcat cccacaggac gatatcgata cgctcgccac ccagttcagt agacaccgcc 1081 tgaacacgcg cgccacgcat acctacgcaa gcacctaccg gatcgatacg tttatcgttg 1141 gttttcaccg cgattttcgc acgagaaccc ggatcgcgag ccgctgcttt aatttcaatc 1201 acttcttcgc cgatttctgg cacttcaata cggaacagtt cgatcagcat ttccggcttg 1261 gaacgagtga cgaacagttg cgcgccacgc gcttccgggc gaacggaata gagcacgcca 1321 cgaacgcggt cgccagggcg gaagttttca cgcggcagca tatcttcgcg caggatcacg 1381 gcttcagcgt tgttgcccag atccagagag atgttgtcgc ggtttacttt tttcaccacg 1441 ccggtgatga tttcaccttc gtgttcacgg aactgatcaa ccaccatcgc acgttcggct 1501 tcacgcactt tctgcacgat aacctgtttt gccgtctggg tagtgatacg gtcaaaggta 1561 acagactcaa tctgatcttc aacgtaatcg cccaggttca ggctttcatc ttcataacgt 1621 gcggcttcaa gggtgatttc cttggtcggc tgggtgactt catcaacaac taaccagcga 1681 cggaaagtgt caaaatcacc gcttttgcga tcgatctgta cgcggacgtc gatctcttgt 1741 tcatattttt tctttgttgc tgtcgccagc gcgctttcca atgcttcgaa aatcttctcg 1801 cgaggtagcg ccttttcatt ggatacggct tcaactacag ccaaaatttc tttgttcata 1861 ctagaatggt gatggtggtg atggctgctg cccatatgta tatctccttc ttaaagttaa 1921 acaaaattat ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta 1981 ttaatttcgc gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca 2041 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag 2101 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc 2161 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg 2221 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag 2281 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag 2341 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc 2401 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac 2461 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc 2521 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc 2581 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa 2641 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg 2701 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat 2761 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat 2821 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg 2881 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta 2941 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa 3001 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc 3061 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg 3121 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg 3181 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc 3241 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag 3301 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc 3361 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg 3421 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta 3481 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag 3541 ctccttccgg tgggcgcggg gcatgactag catgatcgtg ctcctgtcgt tgaggacccg 3601 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac 3661 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg 3721 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg 3781 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa 3841 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg 3901 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag 3961 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga 4021 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca 4081 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg 4141 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga 4201 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc 4261 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg 4321 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca 4381 tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg 4441 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct 4501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 4561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 4621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 4681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 4741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 4801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 4861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 4921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 4981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 5041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg 5101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 5161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 5221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 5281 acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac 5341 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga 5401 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg 5461 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg 5521 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg 5581 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca 5641 tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct 5701 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt 5761 cctgtttgta attgtccttt taacagtgat cgcgtatttc gtctcgctca ggcgcaatca 5821 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct 5881 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc 5941 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt 6001 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac 6061 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat 6121 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta 6181 attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 6241 cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc 6301 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 6361 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 6421 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 6481 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 6541 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 6601 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 6661 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 6721 gtcccattcg cca //
pET-50b(+)载体图谱 pET50b质粒图谱pdf版和pET-50b(+)载体序列 pET50b质粒序列等相关资料下载
pET-50b(+)质粒 pET50b载体应用举例
- 上一篇:pET-49b(+)
- 下一篇:pETDuet-1