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pET-45b(+)

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At a Glance

pET-45b(+), Image provided by Novagen

Type

Expression

Origin of Replication

f1
pBR322

Copy #

~40

Markers

Ampicillin

Link to Sequence

Novagen

Features

N Terminal His Tag
MCS
C Terminal S Tag
lacI
T7 Promoter

Accession Number




Notes

The pET-45b(+) plasmid has the capability to express fusion proteins with an N-terminal His•Tag coding sequence that results in native protein after the purification and cleavage process. The plasmid contains a strong T7lac promoter, an amino-terminal His•Tag coding sequence, and multiple cloning site (MCS) regions which are designed to allow the generation of target proteins with minimal vector-encoded fusion. For inserts that are blunt and in the appropriate reading range, gives direct fusion to the His•Tag sequence in the Pml I cloning site. If a removable amino-terminal His•Tag sequence is required, the MCS includes a PshA I cloning site (GACNNNNGTC). This cloning site overlaps the cleavage site for the enterokinase (EK) protease (AspAspAspAspLys). Cloning that is designed insets into this site recreating the full EK site and allows all amino-terminal vector-encoded sequences to be removed by EK digestion. The optional C-terminal S•Tag coding sequence is compatible with purification, detection, and quantification

History

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Sources

Novagen

References

[1] [2] [3]

  1. Iyer, RS et al. (1991) Quantitation of proteinuria using protein-creatinine ratio in random urine samples. Indian Pediatr 28 463-7 PubMed EcoliWiki page
  2. Sun, G et al. (2007) Structural and functional analyses of the major outer membrane protein of Chlamydia trachomatis. J. Bacteriol. 189 6222-35 PubMed EcoliWiki page
  3. Dai, J et al. (2005) The kinase haspin is required for mitotic histone H3 Thr 3 phosphorylation and normal metaphase chromosome alignment. Genes Dev. 19 472-88 PubMed EcoliWiki page

Sequence

       1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 
      61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 
     121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 
     181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 
     241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 
     301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 
     361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 
     421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttctg gcggcacgat 
     481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 
     541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 
     601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 
     661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 
     721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 
     781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 
     841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 
     901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 
     961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 
    1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 
    1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 
    1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 
    1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 
    1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 
    1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 
    1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 
    1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg 
    1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac 
    1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 
    1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 
    1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 
    1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 
    1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 
    1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 
    1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 
    1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa 
    2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 
    2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 
    2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 
    2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 
    2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 
    2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt 
    2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac 
    2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg 
    2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg 
    2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg 
    2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg 
    2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga 
    2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg 
    2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa 
    2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt 
    2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt 
    3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct 
    3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac 
    3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag 
    3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc 
    3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc 
    3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc 
    3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 
    3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 
    3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt 
    3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg 
    3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc 
    3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat 
    3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt 
    3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc 
    3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc 
    3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag 
    3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt 
    4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat 
    4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag 
    4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac 
    4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg 
    4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg 
    4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt 
    4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc 
    4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact 
    4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg 
    4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga 
    4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc 
    4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt 
    4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg 
    4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc 
    4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag 
    4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac 
    4981 gtgggtaccg gttcgaatga tgacgacgac aagagtccgg atcccaattg ggagctcgtg 
    5041 tacacggcgc gcctgcaggt cgacaagctt gcggccgcac tcgagtctgg taaagaaacc 
    5101 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc 
    5161 taggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc 
    5221 ttgaggggtt ttttgctgaa aggaggaact atatccggat