pET-47b(+)
At a Glance
<protect>
Type |
Expression |
---|---|
Origin of Replication |
f1 |
Copy # |
~40 |
Markers |
Kanamycin |
Link to Sequence | |
Features |
T7 Promoter
|
Accession Number |
|
edit table |
</protect>
Notes
The pET-47b(+) vector expresses fusion protein with a N-terminal His•Tag® coding sequence that can be cleaved with the human rhinovirus (HRV) 3C protease. This protease cleaves the sequence LEVLFQ↓GP specifically as well as maintinas activity at low temperatures. In addition, pET-47b(+) contains an optional C-Terminal thrombin recognition site, in addition to an STag coding sequence, compatible with purification, detection, and quantification. The plasmid also contains a strong T7lac promoter, an optimized RBS, the coding sequence for the HRV 3C protease cleavage site (LeuGluValLeuPheGlnGlyPro), and a multiple cloning site that contains restriction enzyme sites. Single-stranded sequencing should be performed using the AS S•Tag 18mer Primer
History
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Sources
References
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Sequence
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