pET-27b(+)
At a Glance
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pET-27b(+), Image provided by Novagen
<protect>
| Type |
Expression |
|---|---|
| Origin of Replication |
f1 |
| Copy # |
~40 |
| Markers |
Kanamycin |
| Link to Sequence | |
| Features |
HSV Tag |
| Accession Number |
|
| edit table |
</protect>
Notes
The pET-27b(+) vector carries an N-terminal pelB signal sequence for potential periplasmic localization, in addition to an optional C-terminal HSV•Tag and His•Tag sequence. This vector in comparison to pET-25b(+) differs only by its selectable marker (kanamycin vs. ampicillin resistance). The T7 expression region is reversed on the circular map due to the sequence numbering 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, the T7 terminator primers should be used when performing single-stranded sequencing.
History
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Sources
References
- ↑ Cellitti, J et al. (2007) Exploring subdomain cooperativity in T4 lysozyme I: structural and energetic studies of a circular permutant and protein fragment. Protein Sci. 16 842-51 PubMed
- ↑ Flanigan, A & Gardner, JF (2007) Interaction of the Gifsy-1 Xis protein with the Gifsy-1 attP sequence. J. Bacteriol. 189 6303-11 PubMed
- ↑ Johnson, RJ et al. (2007) Genetic selection reveals the role of a buried, conserved polar residue. Protein Sci. 16 1609-16 PubMed
- ↑ Liu, D et al. (2005) Cyclic changes in the affinity of protein-DNA interactions drive the progression and regulate the outcome of the Tn10 transposition reaction. Nucleic Acids Res. 33 1982-92 PubMed
- ↑ Dai, KS & Liew, CC (2001) A novel human striated muscle RING zinc finger protein, SMRZ, interacts with SMT3b via its RING domain. J. Biol. Chem. 276 23992-9 PubMed
- ↑ Brandt, JJ et al. (2000) Analysis of three overexpression systems for VanX, the Zinc(II) dipeptidase required for high-level vancomycin resistance in bacteria. Protein Expr. Purif. 20 300-7 PubMed
- ↑ Kenny, B et al. (1997) Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91 511-20 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 atgaacaata aaactgtctg cttacataaa
4021 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct ctaggccgcg
4081 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
4141 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
4201 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
4261 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
4321 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
4381 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
4441 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
4501 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
4561 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
4621 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
4681 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
4741 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
4801 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaagaatt
4861 aattcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
4921 gcacatttcc ccgaaaagtg ccacctgaaa ttgtaaacgt taatattttg ttaaaattcg
4981 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
5041 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
5101 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
5161 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
5221 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
5281 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
5341 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
5401 cgtcccattc gcca
