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pET-27b(+)

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At a Glance

pET-27b(+), Image provided by Novagen

Type

Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Kanamycin

Link to Sequence

Novagen

Features

HSV Tag
His Tag
T7 Promoter
MCS
lacI

Accession Number




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

You can help EcoliWiki by writing about the history of this plasmid.

Sources

Novagen

References

[1] [2] [3] [4] [5] [6] [7]

  1. 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 EcoliWiki page
  2. Flanigan, A & Gardner, JF (2007) Interaction of the Gifsy-1 Xis protein with the Gifsy-1 attP sequence. J. Bacteriol. 189 6303-11 PubMed EcoliWiki page
  3. Johnson, RJ et al. (2007) Genetic selection reveals the role of a buried, conserved polar residue. Protein Sci. 16 1609-16 PubMed EcoliWiki page
  4. 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 EcoliWiki page
  5. 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 EcoliWiki page
  6. 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 EcoliWiki page
  7. Kenny, B et al. (1997) Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91 511-20 PubMed EcoliWiki page

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