pET-42a(+)
At a Glance
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pET-42a(+), Image provided by Novagen
<protect>
| Type |
Cloning, Expression |
|---|---|
| Origin of Replication |
f1 |
| Copy # |
~40 |
| Markers |
Kanamycin |
| Link to Sequence | |
| Features |
T7 Promoter |
| Accession Number |
|
| edit table |
</protect>
Notes
The pET-42a(+) has the popular GST fusion tag for enhanced production and solubility.
The pET-42 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST•Tag™ protein. The T7 expression region is reversed on the vector map due to the sequencing numbered by the pBR322 convention. 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. When cloning into the PshaI site, the vector encoded sequence can be removed completely followed by the cleaving of the GST fusion protein with Factor Xa.
History
You can help EcoliWiki by writing about the history of this plasmid.
Sources
References
[1] [2] [3] [4] [5] [6] [7] [8] [9] [10]
- ↑ Kulman, JD et al. (2007) Proline-rich Gla protein 2 is a cell-surface vitamin K-dependent protein that binds to the transcriptional coactivator Yes-associated protein. Proc. Natl. Acad. Sci. U.S.A. 104 8767-72 PubMed
- ↑ Beernink, PT et al. (2005) Specificity of protein interactions mediated by BRCT domains of the XRCC1 DNA repair protein. J. Biol. Chem. 280 30206-13 PubMed
- ↑ Chellappan, P et al. (2005) MicroRNA-binding viral protein interferes with Arabidopsis development. Proc. Natl. Acad. Sci. U.S.A. 102 10381-6 PubMed
- ↑ Fotso, P et al. (2005) Cog1p plays a central role in the organization of the yeast conserved oligomeric Golgi complex. J. Biol. Chem. 280 27613-23 PubMed
- ↑ Li, L et al. (2005) Clarification of cinnamoyl co-enzyme A reductase catalysis in monolignol biosynthesis of Aspen. Plant Cell Physiol. 46 1073-82 PubMed
- ↑ Liedtke, CM et al. (2005) Role for protein phosphatase 2A in the regulation of Calu-3 epithelial Na+-K+-2Cl-, type 1 co-transport function. J. Biol. Chem. 280 25491-8 PubMed
- ↑ Madsen, KL et al. (2005) Molecular determinants for the complex binding specificity of the PDZ domain in PICK1. J. Biol. Chem. 280 20539-48 PubMed
- ↑ Srivastava, SK et al. (2005) NAD+-dependent DNA Ligase (Rv3014c) from Mycobacterium tuberculosis. Crystal structure of the adenylation domain and identification of novel inhibitors. J. Biol. Chem. 280 30273-81 PubMed
- ↑ Yang, HY et al. (2005) Kinesin-1 mediates translocation of the meiotic spindle to the oocyte cortex through KCA-1, a novel cargo adapter. J. Cell Biol. 169 447-57 PubMed
- ↑ Zong, X et al. (2005) A novel mechanism of modulation of hyperpolarization-activated cyclic nucleotide-gated channels by Src kinase. J. Biol. Chem. 280 34224-32 PubMed
Sequence
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
241 ggatccccga tatcccatgg acccgcgtcc ctcaataccg gagccaccac cggtacccag
301 atctgggctg tccatgtgct ggcgttcgaa tttagcagca gcggtttctt tcataccaat
361 tgcagtacta ccgcgtggca ccagacccgc ggagtgatgg tgatggtgat gaccagaacc
421 actagttgaa ccatccgatt ttggaggatg gtcgccacca ccaaacgtgg cttgccagcc
481 ctgcaaaggc catgctatat acttgctgga tttcaagtac ttatcaattt gtgggatagc
541 ttcaatacgt tttttaaaac aaactaattt tgggaacgca tccaggcaca ttgggtccat
601 gtataaaaca acatcaagag cgtcatacaa catgaagtca ggatgggtta catgatcacc
661 atttaaatat gttttatgac ataaacgatc ttcgaacatt ttcagcattt caggtagctt
721 gctaagaaaa tcaactttga gagtttcaaa gtctttacta tatgcaattc tcgaaacacc
781 gtatctaata tccaaaaccg ctccttcaag cattgaaatc tctgcacgct cttttggaca
841 accacccaac atgttgtgct tgtcagctat ataacgtatg atggccatag actgtgttaa
901 tttaacatca ccatcaatat aataaggaag attgggaaac tccaaaccca attcaaactt
961 tttgtttcgc catttatcac cttcatcgcg ctcatacaaa tgctcttcat atttttcttc
1021 aagatattcc aaaagaagtc gagtgggttg cacaaggccc ttaattttcc aataacctag
1081 tataggggac atatgtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
1141 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatcg
1201 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
1261 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
1321 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
1381 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
1441 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
1501 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
1561 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
1621 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
1681 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
1741 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
1801 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
1861 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
1921 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
1981 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
2041 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
2101 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
2161 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
2221 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
2281 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
2341 taccgagtcc gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga
2401 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
2461 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
2521 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
2581 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
2641 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
2701 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
2761 tgactagcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
2821 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
2881 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
2941 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
3001 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
3061 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
3121 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
3181 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
3241 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
3301 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
3361 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
3421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
3481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
3541 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
3601 gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
3661 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
3721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
3781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
3841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
3901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
3961 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
4021 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
4081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
4141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
4201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
4261 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
4321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
4381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
4441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
4501 ttggtcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
4561 ccatattcaa cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga
4621 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
4681 attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
4741 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
4801 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
4861 cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
4921 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
4981 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
5041 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
5101 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
5161 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
5221 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
5281 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
5341 gtttcatttg atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg
5401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
5461 ctgaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
5521 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
5581 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
5641 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
5701 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
5761 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
5821 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
5881 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
