| 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
//
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