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pET47b载体基本信息
| EMD Biosciences (Novagen) |
| pET47b, pET 47b |
| 大肠杆菌蛋白表达 |
| 高 |
| 多克隆位点，限制性内切酶 |
| 5203 bp |
| T7: 5'-TAATACGACTCACTATAGGG-3' |
| T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
| N-His, C-S, C-Thrombin |
| 卡那 |
载体含有HRV 3C蛋白酶切位点，可以在低温条件下将多余的融合表达蛋白切除。载体上还有C端Thrombin蛋白酶切位点可以使用。 |
| 71461-3 |
| 瞬时表达 Transient |
| 组成型 Constitutive |
| 非病毒 |
pET47b载体质粒图谱和多克隆位点信息
pET47b载体简介
pET47b载体携带有N端His纯化标签，紧跟着有一个HRV 3C蛋白酶切位点。这个HRV 3C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。
pET-47b(+)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。
pET47b载体序列
LOCUS pET-47b(+) 5203 bp DNA Circular SYN
DEFINITION pET-47b(+)
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..5203
/organism="pET-47b(+)"
/mol_type="other DNA"
terminator complement(1..120)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
CDS 345..1094
/label="ORF frame 3"
misc_feature complement(374..390)
/label="T7_transl_en_RBS"
misc_feature complement(408..435)
/label="lacO"
promoter complement(435..453)
/label="T7_promoter"
gene 489..752
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(541..560)
/label="pBRrevBam_primer"
misc_feature 835..1926
/label="lacI"
CDS 967..1926
/label="ORF frame 1"
misc_feature 2498..2689
/label="ROP"
misc_feature complement(2683..2705)
/label="pGEX_3_primer"
rep_origin complement(3104..3723)
/label="pBR322_origin"
gene 3829..4644
/label="KanR2"
/gene="KanR2"
CDS 3829..4644
/label="ORF frame 1"
rep_origin complement(4869..5175)
/label="f1_origin"
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 gtggtgatgt
361 gccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
421 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga
481 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
541 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
601 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
661 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
721 gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg
781 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
841 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
901 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
961 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
1021 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
1081 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
1141 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
1201 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
1261 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
1321 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
1381 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
1441 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
1501 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
1561 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
1621 tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc
1681 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
1741 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
1801 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
1861 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
1921 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc
1981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactag
2041 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
2101 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
2161 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
2221 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
2281 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
2341 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
2401 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
2461 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
2521 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
2581 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
2641 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
2701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
2761 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
2821 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
2881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3541 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
3781 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
3841 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
3901 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
3961 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
4021 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
4081 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
4141 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
4201 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagtgat
4261 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
4321 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
4381 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
4441 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
4501 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
4561 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
4621 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
4681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
4741 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
4801 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
4861 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
4921 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
4981 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
5041 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
5101 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
5161 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//
DEFINITION pET-47b(+)
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..5203
/organism="pET-47b(+)"
/mol_type="other DNA"
terminator complement(1..120)
/label="T7_terminator"
misc_feature 69..87
/label="T7_Terminal_primer"
CDS 345..1094
/label="ORF frame 3"
misc_feature complement(374..390)
/label="T7_transl_en_RBS"
misc_feature complement(408..435)
/label="lacO"
promoter complement(435..453)
/label="T7_promoter"
gene 489..752
/label="tet (300 - 563)"
/gene="tet (300 - 563)"
misc_feature complement(541..560)
/label="pBRrevBam_primer"
misc_feature 835..1926
/label="lacI"
CDS 967..1926
/label="ORF frame 1"
misc_feature 2498..2689
/label="ROP"
misc_feature complement(2683..2705)
/label="pGEX_3_primer"
rep_origin complement(3104..3723)
/label="pBR322_origin"
gene 3829..4644
/label="KanR2"
/gene="KanR2"
CDS 3829..4644
/label="ORF frame 1"
rep_origin complement(4869..5175)
/label="f1_origin"
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 gtggtgatgt
361 gccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
421 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga
481 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
541 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
601 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
661 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
721 gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg
781 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
841 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
901 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
961 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
1021 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
1081 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
1141 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
1201 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
1261 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
1321 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
1381 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
1441 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
1501 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
1561 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
1621 tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc
1681 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
1741 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
1801 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
1861 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
1921 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc
1981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactag
2041 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
2101 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
2161 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
2221 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
2281 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
2341 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
2401 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
2461 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
2521 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
2581 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
2641 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
2701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
2761 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
2821 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
2881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3541 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
3781 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
3841 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
3901 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
3961 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
4021 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
4081 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
4141 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
4201 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagtgat
4261 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
4321 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
4381 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
4441 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
4501 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
4561 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
4621 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
4681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
4741 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
4801 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
4861 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
4921 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
4981 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
5041 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
5101 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
5161 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca
//
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