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pET-49b(+)

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At a Glance

PET-49b(+).jpg Image provided by Novagen

Type

Cloning, Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Ampicillin

Link to Sequence

Novagen

Features

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

Accession Number




Notes

The pET-49b(+) vector is designed for cloning and high-level expression of target proteins fused with the 220 aa schistosomal glutathione-S-transferase (GST•Tag™) protein that is cleavable with the human rhinovirus (HRV) 3C protease . The GST tag, used as fusion when expressing proteins in E. Coli, has been reported to enhance the production and in certain situations the solubility of its fusion partners. When expressed in a soluble, properly folded form, GST•Tag fusion proteins can be purified with immobilized glutathione. The plasmid contains a strong T7lac promoter, optimized RBS, the coding sequence for the HRV 3C protease cleavage site (LeuGluValLeuPheGlnGlyPro) for N-terminal fusion tag removal and a multiple cloning site that contains restriction enzyme sites. An optional C-terminal S•Tag™ coding sequence is compatible with purification, detection, and quantification. Single-stranded sequencing should be performed using the AS S•Tag 18mer

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 atggtgatga 
     361 ccagaaccac tagatggagt aggagtagga ggattgttat tagaaccgcc accaccacta 
     421 gttgaaccat ccgattttgg aggatggtcg ccaccaccaa acgtggcttg ccagccctgc 
     481 aaaggccatg ctatatactt gctggatttc aagtacttat caatttgtgg gatagcttca 
     541 atacgttttt taaaacaaac taattttggg aacgcatcca ggcacattgg gtccatgtat 
     601 aaaacaacat caagagcgtc atacaacatg aagtcaggat gggttacatg atcaccattt 
     661 aaatatgttt tatgacataa acgatcttcg aacattttca gcatttcagg tagcttgcta 
     721 agaaaatcaa ctttgagagt ttcaaagtct ttactatatg caattctcga aacaccgtat 
     781 ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg cacgctcttt tggacaacca 
     841 cccaacatgt tgtgcttgtc agctatataa cgtatgatgg ccatagactg tgttaattta 
     901 acatcaccat caatataata aggaagattg ggaaactcca aacccaattc aaactttttg 
     961 tttcgccatt tatcaccttc atcgcgctca tacaaatgct cttcatattt ttcttcaaga 
    1021 tattccaaaa gaagtcgagt gggttgcaca aggcccttaa ttttccaata acctagtata 
    1081 ggggacatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt 
    1141 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct 
    1201 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg 
    1261 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga 
    1321 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca 
    1381 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg 
    1441 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa 
    1501 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg 
    1561 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag 
    1621 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg 
    1681 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc 
    1741 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa 
    1801 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg 
    1861 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc 
    1921 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct 
    1981 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt 
    2041 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt 
    2101 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg 
    2161 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc 
    2221 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt 
    2281 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc 
    2341 gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac 
    2401 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa 
    2461 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg 
    2521 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct 
    2581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc 
    2641 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg 
    2701 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac 
    2761 tagcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg 
    2821 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc 
    2881 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac 
    2941 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct 
    3001 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt 
    3061 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg 
    3121 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat 
    3181 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac 
    3241 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa 
    3301 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga 
    3361 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 
    3421 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 
    3481 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga 
    3541 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac 
    3601 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct 
    3661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 
    3721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 
    3781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 
    3841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 
    3901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 
    3961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 
    4021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 
    4081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 
    4141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 
    4201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 
    4261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 
    4321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 
    4381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 
    4441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 
    4501 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat 
    4561 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta 
    4621 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg 
    4681 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat 
    4741 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc 
    4801 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc 
    4861 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg 
    4921 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagt 
    4981 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg 
    5041 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat 
    5101 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac 
    5161 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca 
    5221 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta 
    5281 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt 
    5341 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg 
    5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga 
    5461 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 
    5521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 
    5581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 
    5641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 
    5701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 
    5761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 
    5821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 
    5881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca