pET-25b(+)
At a Glance
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pET-25b(+), Image made with PlasMapper
<protect>
| Type |
Expression |
|---|---|
| Origin of Replication |
f1, pBR322 |
| Copy # |
~40 |
| Markers |
Ampicillin |
| Link to Sequence | |
| Features |
T7 Promoter |
| Accession Number |
|
| edit table |
</protect>
Notes
The pET-25b(+) vecotor has potential periplasmic localiztion. For this reason the plasmid carries an N-terminal perlB signal sequence. there are alos optional C-terminal HSV tags and His tag sequences. Helper phage are able to infect and produce virions which contain single stranded DNA that corresponds with the coding strand. This happens because the f1 origin is oriented in a specific region to allow this. Single-stranded sequencing should be performed by the t& terminator primer.
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]
- ↑ Bidwell, GL 3rd & Raucher, D (2005) Application of thermally responsive polypeptides directed against c-Myc transcriptional function for cancer therapy. Mol. Cancer Ther. 4 1076-85 PubMed
- ↑ Kostlánová, N et al. (2005) The fucose-binding lectin from Ralstonia solanacearum. A new type of beta-propeller architecture formed by oligomerization and interacting with fucoside, fucosyllactose, and plant xyloglucan. J. Biol. Chem. 280 27839-49 PubMed
- ↑ Zhou, H et al. (2005) Contributions of disulfide bonds in a nested pattern to the structure, stability, and biological functions of endostatin. J. Biol. Chem. 280 11303-12 PubMed
- ↑ Trabbic-Carlson, K et al. (2004) Effect of protein fusion on the transition temperature of an environmentally responsive elastin-like polypeptide: a role for surface hydrophobicity? Protein Eng. Des. Sel. 17 57-66 PubMed
- ↑ Deckert, PM et al. (2003) A33scFv-cytosine deaminase: a recombinant protein construct for antibody-directed enzyme-prodrug therapy. Br. J. Cancer 88 937-9 PubMed
- ↑ Cai, S & Exton, JH (2001) Determination of interaction sites of phospholipase D1 for RhoA. Biochem. J. 355 779-85 PubMed
- ↑ Cooper, KW & Baneyx, F (2001) Escherichia coli FtsH (HflB) degrades a membrane-associated TolAI-II-beta-lactamase fusion protein under highly denaturing conditions. Protein Expr. Purif. 21 323-32 PubMed
- ↑ West, MW et al. (1999) De novo amyloid proteins from designed combinatorial libraries. Proc. Natl. Acad. Sci. U.S.A. 96 11211-6 PubMed
- ↑ Murley, LL & Grindley, ND (1998) Architecture of the gamma delta resolvase synaptosome: oriented heterodimers identity interactions essential for synapsis and recombination. Cell 95 553-62 PubMed
- ↑ Zhou, P et al. (1998) Cells transfected with a non-neutralizing antibody gene are resistant to HIV infection: targeting the endoplasmic reticulum and trans-Golgi network. J. Immunol. 160 1489-96 PubMed
Sequence
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gacatcctcg gggtcttccg
181 gggcgagttc tggctggcta gcccgtttga tctcgagtgc ggccgcaagc ttgtcgacgg
241 agctcgaatt cggatccgaa ttaattccga tatccatggc catcgccggc tgggcagcga
301 ggagcagcag accagcagca gcggtcggca gcaggtattt catatgtata tctccttctt
361 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
421 gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc
481 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
541 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
601 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
661 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
721 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
781 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
841 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
901 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
961 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
1021 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
1081 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
1141 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
1201 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
1261 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
1321 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
1381 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
1441 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
1501 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
1561 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
1621 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
1681 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
1741 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
1801 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
1861 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
1921 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag
1981 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
2041 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
2101 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
2161 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
2221 ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc
2281 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
2341 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
2401 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
2461 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
2521 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
2581 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
2641 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
2701 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
2761 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
2821 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
2881 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
3001 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
3061 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
3121 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
3481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
3961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
4021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
4081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
4141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
4201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
4261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
4321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
4381 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
4441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
4501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
4561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
4621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
4681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
4741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
4801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
4861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
4921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
4981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
5041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt
5101 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
5161 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
5221 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
5281 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
5341 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
5401 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
5461 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
5521 ccgctacagg gcgcgtccca ttcgcca
