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pET-47b(+)

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At a Glance

pET-47b(+), Image provided by Novagen

Type

Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

T7 Promoter
MCS
N Terminal His Tag
C Terminal S Tag
lacI


Accession Number




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

Novagen

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