pET-49b(+)
At a Glance
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PET-49b(+).jpg Image provided by Novagen
<protect>
| Type |
Cloning, Expression |
|---|---|
| Origin of Replication |
f1 |
| Copy # |
~40 |
| Markers |
Ampicillin |
| Link to Sequence | |
| Features |
T7 Promoter |
| Accession Number |
|
| edit table |
</protect>
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
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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 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
