pET-29b(+)
At a Glance
<protect>
Type |
Cloning, Expression |
---|---|
Origin of Replication |
f1 |
Copy # |
~40 |
Markers |
Kanamycin |
Link to Sequence | |
Features |
T7 Promoter |
Accession Number | |
edit table |
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Notes
The pET-29b(+) vector carries an N-terminal S•Tag™/thrombin configuration including an optional C-terminal His•Tag® sequence. The T7 expression region is reversed on the vector map due to the sequencing numbered by the pBR322 convention. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is included. The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, T7 terminator primer should be used for single-stranded sequencing.
History
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Sources
References
[1] [2] [3] [4] [5] [6] [7] [8] [9] [10]
- ↑ Fischbach, MA et al. (2007) Directed evolution can rapidly improve the activity of chimeric assembly-line enzymes. Proc. Natl. Acad. Sci. U.S.A. 104 11951-6 PubMed
- ↑ Hsu, IaW et al. (2005) Phosphorylation of Y14 modulates its interaction with proteins involved in mRNA metabolism and influences its methylation. J. Biol. Chem. 280 34507-12 PubMed
- ↑ Jilaveanu, LB et al. (2005) Dimeric SecA is essential for protein translocation. Proc. Natl. Acad. Sci. U.S.A. 102 7511-6 PubMed
- ↑ Korepanova, A et al. (2005) Cloning and expression of multiple integral membrane proteins from Mycobacterium tuberculosis in Escherichia coli. Protein Sci. 14 148-58 PubMed
- ↑ Müllegger, J et al. (2005) Engineering of a thioglycoligase: randomized mutagenesis of the acid-base residue leads to the identification of improved catalysts. Protein Eng. Des. Sel. 18 33-40 PubMed
- ↑ Nam, JO et al. (2005) Regulation of tumor angiogenesis by fastatin, the fourth FAS1 domain of betaig-h3, via alphavbeta3 integrin. Cancer Res. 65 4153-61 PubMed
- ↑ Zhang, W et al. (2005) Biophysical and kinetic characterization of HemAT, an aerotaxis receptor from Bacillus subtilis. Biophys. J. 88 2801-14 PubMed
- ↑ Zito, CR et al. (2005) Role of a conserved glutamate residue in the Escherichia coli SecA ATPase mechanism. J. Biol. Chem. 280 14611-9 PubMed
- ↑ Kanaya, E et al. (1999) Characterization of the transcriptional activator CBF1 from Arabidopsis thaliana. Evidence for cold denaturation in regions outside of the DNA binding domain. J. Biol. Chem. 274 16068-76 PubMed
- ↑ Huang, B et al. () NMR structure and mutagenesis of the Fas (APO-1/CD95) death domain. Nature 384 638-41 PubMed
Sequence
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt 181 cgacggagct cgaattcgga tccgatatcg ccatggaacc gcgtggcacc agggtaccca 241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcatatgta 301 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa 361 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc 421 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 481 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 541 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 601 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 661 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct 721 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac 781 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg 841 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg 901 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc 961 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga 1021 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg 1081 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga 1141 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg 1201 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc 1261 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg 1321 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata 1381 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca 1441 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc 1501 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc 1561 gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata 1621 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc 1681 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac 1741 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg 1801 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 1861 aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc gatgcccttg 1921 agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca 1981 cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc 2041 attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta 2101 ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc 2161 ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat gatcgtgctc 2221 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca 2281 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca 2341 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc 2401 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct 2461 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc 2521 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca 2581 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga 2641 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc 2701 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat 2761 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc 2821 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 2881 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 2941 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact 3001 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata tgcggtgtga 3061 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct 3121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 3181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 3241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 3301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 3361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 3421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 3481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 3541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 3601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 3661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 3721 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 3781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 3841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 3901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga acaataaaac 3961 tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt 4021 cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 4081 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg 4141 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 4201 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 4261 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca gcattccagg 4321 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc 4381 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 4441 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 4501 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct 4561 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 4621 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc 4681 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc 4741 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg 4801 agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta gaaaaataaa 4861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt aaacgttaat 4921 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 4981 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 5041 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 5101 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 5161 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 5221 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 5281 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 5341 gcgccgctac agggcgcgtc ccattcgcca