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pET-11b

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At a Glance

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pET-11b, Image provided by Agilent-Genomics

Type

Expression

Origin of Replication

pBR322

Copy #
Markers

Ampicillin

Link to Sequence

Novagen

Features

T7 promoter
lac operator
ribosome binding site
Nde I cloning site
BamH I cloning site
lacI repressor ORF

Accession Number




Notes

In the pET system the sequence of interest is transformed into E. coli after being transcribed downstream from the T7 promoter and the gene 10 ribosome binding site. One can use IPTG induction of a chromosomally integrated cassette or infection with a polymerase-expressing bacteriophage lamba CE6 to achieve protien expression. After induction to E. coli the very active polymerase cause a high nimber of transcipts. Due to this phenomenon and high efficiency of translation, the target protein may become the majority of the cellular protein in only a matter of hours. [1]

History

The pET-11 plasmids are the improved form of the pET-3 plasmids. The lac operator has been placed in between the T7 promoter and the translation initiation sequences to produce a higher level of regulation and control. IPTG has to be present or the plasmid results in repression. These plasmids also include the lacIq gene to provide adequate levels of LacI repressor protiens. This shuts off the T7 polymerase gene expression along with the T7 promoter transcription.[2]

Sources

Agilent Genomics

Novagen

References

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See Help:References for how to manage references in EcoliWiki.

  1. Bonin, C. "PNAS,Vol 94". 1996. Page 2085-9090.
  2. Bonin, C. "PNAS,Vol 94". 1996. Page 2085-9090

Sequence

THE SEQUENCE PROVIDED IS NOT COMPATIBLE WITH RESTRICTION MAPS OR VECTOR INFORMATION OBTAINED PRIOR TO DECEMBER, 2001.

   1  TAATACGACT CACTATAGGG GAATTGTGAG CGGATAACAA TTCCCCTCTA  
  51  GAAATAATTT TGTTTAACTT TAAGAAGGAG ATATACATAT GGCTAGCATG  
 101  ACTGGTGGAC AGCAAATGGG TCGGGATCCG GCTGCTAACA AAGCCCGAAA  
 151  GGAAGCTGAG TTGGCTGCTG CCACCGCTGA GCAATAACTA GCATAACCCC  
 201  TTGGGGCCTC TAAACGGGTC TTGAGGGGTT TTTTGCTGAA AGGAGGAACT  
 251  ATATCCGGAT ATCCCGCAAG AGGCCCGGCA GTACCGGCAT AACCAAGCCT  
 301  ATGCCTACAG CATCCAGGGT GACGGTGCCG AGGATGACGA TGAGCGCATT  
 351  GTTAGATTTC ATACACGGTG CCTGACTGCG TTAGCAATTT AACTGTGATA  
 401  AACTACCGCA TTAAAGCTTA TCGATGATAA GCTGTCAAAC ATGAGAATTC  
 451  TTGAAGACGA AAGGGCCTCG TGATACGCCT ATTTTTATAG GTTAATGTCA  
 501  TGATAATAAT GGTTTCTTAG ACGTCAGGTG GCACTTTTCG GGGAAATGTG  
 551  CGCGGAACCC CTATTTGTTT ATTTTTCTAA ATACATTCAA ATATGTATCC  
 601  GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT TGAAAAAGGA  
 651  AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG  
 701  GCATTTTGCC TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA  
 751  AGATGCTGAA GATCAGTTGG GTGCACGAGT GGGTTACATC GAACTGGATC  
 801  TCAACAGCGG TAAGATCCTT GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA  
 851  ATGATGAGCA CTTTTAAAGT TCTGCTATGT GGCGCGGTAT TATCCCGTGT  
 901  TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT TCTCAGAATG  
 951  ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG  
1001  ACAGTAAGAG AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC  
1051  GGCCAACTTA CTTCTGACAA CGATCGGAGG ACCGAAGGAG CTAACCGCTT  
1101  TTTTGCACAA CATGGGGGAT CATGTAACTC GCCTTGATCG TTGGGAACCG  
1151  GAGCTGAATG AAGCCATACC AAACGACGAG CGTGACACCA CGATGCCTGC  
1201  AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA CTACTTACTC  
1251  TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA  
1301  GGACCACTTC TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA  
1351  ATCTGGAGCC GGTGAGCGTG GGTCTCGCGG TATCATTGCA GCACTGGGGC  
1401  CAGATGGTAA GCCCTCCCGT ATCGTAGTTA TCTACACGAC GGGGAGTCAG  
1451  GCAACTATGG ATGAACGAAA TAGACAGATC GCTGAGATAG GTGCCTCACT  
1501  GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT ATACTTTAGA  
1551  TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT  
1601  TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG  
1651  AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT  
1701  TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG  
1751  GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC  
1801  TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT  
1851  AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT  
1901  CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT  
1951  TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG  
2001  GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC  
2051  ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC  
2101  CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG  
2151  GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT  
2201  CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC  
2251  GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC  
2301  GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATGTTCTT TCCTGCGTTA  
2351  TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC  
2401  CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG  
2451  CGGAAGAGCG CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT  
2501  TCACACCGCA TATATGGTGC ACTCTCAGTA CAATCTGCTC TGATGCCGCA  
2551  TAGTTAAGCC AGTATACACT CCGCTATCGC TACGTGACTG GGTCATGGCT  
2601  GCGCCCCGAC ACCCGCCAAC ACCCGCTGAC GCGCCCTGAC GGGCTTGTCT  
2651  GCTCCCGGCA TCCGCTTACA GACAAGCTGT GACCGTCTCC GGGAGCTGCA  
2701  TGTGTCAGAG GTTTTCACCG TCATCACCGA AACGCGCGAG GCAGCTGCGG  
2751  TAAAGCTCAT CAGCGTGGTC GTGAAGCGAT TCACAGATGT CTGCCTGTTC  
2801  ATCCGCGTCC AGCTCGTTGA GTTTCTCCAG AAGCGTTAAT GTCTGGCTTC  
2851  TGATAAAGCG GGCCATGTTA AGGGCGGTTT TTTCCTGTTT GGTCACTGAT  
2901  GCCTCCGTGT AAGGGGGATT TCTGTTCATG GGGGTAATGA TACCGATGAA  
2951  ACGAGAGAGG ATGCTCACGA TACGGGTTAC TGATGATGAA CATGCCCGGT  
3001  TACTGGAACG TTGTGAGGGT AAACAACTGG CGGTATGGAT GCGGCGGGAC  
3051  CAGAGAAAAA TCACTCAGGG TCAATGCCAG CGCTTCGTTA ATACAGATGT  
3101  AGGTGTTCCA CAGGGTAGCC AGCAGCATCC TGCGATGCAG ATCCGGAACA  
3151  TAATGGTGCA GGGCGCTGAC TTCCGCGTTT CCAGACTTTA CGAAACACGG  
3201  AAACCGAAGA CCATTCATGT TGTTGCTCAG GTCGCAGACG TTTTGCAGCA  
3251  GCAGTCGCTT CACGTTCGCT CGCGTATCGG TGATTCATTC TGCTAACCAG  
3301  TAAGGCAACC CCGCCAGCCT AGCCGGGTCC TCAACGACAG GAGCACGATC  
3351  ATGCGCACCC GTGGCCAGGA CCCAACGCTG CCCGAGATGC GCCGCGTGCG  
3401  GCTGCTGGAG ATGGCGGACG CGATGGATAT GTTCTGCCAA GGGTTGGTTT  
3451  GCGCATTCAC AGTTCTCCGC AAGAATTGAT TGGCTCCAAT TCTTGGAGTG  
3501  GTGAATCCGT TAGCGAGGTG CCGCCGGCTT CCATTCAGGT CGAGGTGGCC  
3551  CGGCTCCATG CACCGCGACG CAACGCGGGG AGGCAGACAA GGTATAGGGC  
3601  GGCGCCTACA ATCCATGCCA ACCCGTTCCA TGTGCTCGCC GAGGCGGCAT  
3651  AAATCGCCGT GACGATCAGC GGTCCAGTGA TCGAAGTTAG GCTGGTAAGA  
3701  GCCGCGAGCG ATCCTTGAAG CTGTCCCTGA TGGTCGTCAT CTACCTGCCT  
3751  GGACAGCATG GCCTGCAACG CGGGCATCCC GATGCCGCCG GAAGCGAGAA  
3801  GAATCATAAT GGGGAAGGCC ATCCAGCCTC GCGTCGCGAA CGCCAGCAAG  
3851  ACGTAGCCCA GCGCGTCGGC CGCCATGCCG GCGATAATGG CCTGCTTCTC  
3901  GCCGAAACGT TTGGTGGCGG GACCAGTGAC GAAGGCTTGA GCGAGGGCGT  
3951  GCAAGATTCC GAATACCGCA AGCGACAGGC CGATCATCGT CGCGCTCCAG  
4001  CGAAAGCGGT CCTCGCCGAA AATGACCCAG AGCGCTGCCG GCACCTGTCC  
4051  TACGAGTTGC ATGATAAAGA AGACAGTCAT AAGTGCGGCG ACGATAGTCA  
4101  TGCCCCGCGC CCACCGGAAG GAGCTGACTG GGTTGAAGGC TCTCAAGGGC  
4151  ATCGGTCGAG ATCCCGGTGC CTAATGAGTG AGCTAACTTA CATTAATTGC  
4201  GTTGCGCTCA CTGCCCGCTT TCCAGTCGGG AAACCTGTCG TGCCAGCTGC  
4251  ATTAATGAAT CGGCCAACGC GCGGGGAGAG GCGGTTTGCG TATTGGGCGC  
4301  CAGGGTGGTT TTTCTTTTCA CCAGTGAGAC GGGCAACAGC TGATTGCCCT  
4351  TCACCGCCTG GCCCTGAGAG AGTTGCAGCA AGCGGTCCAC GCTGGTTTGC  
4401  CCCAGCAGGC GAAAATCCTG TTTGATGGTG GTTAACGGCG GGATATAACA  
4451  TGAGCTGTCT TCGGTATCGT CGTATCCCAC TACCGAGATA TCCGCACCAA  
4501  CGCGCAGCCC GGACTCGGTA ATGGCGCGCA TTGCGCCCAG CGCCATCTGA  
4551  TCGTTGGCAA CCAGCATCGC AGTGGGAACG ATGCCCTCAT TCAGCATTTG  
4601  CATGGTTTGT TGAAAACCGG ACATGGCACT CCAGTCGCCT TCCCGTTCCG  
4651  CTATCGGCTG AATTTGATTG CGAGTGAGAT ATTTATGCCA GCCAGCCAGA  
4701  CGCAGACGCG CCGAGACAGA ACTTAATGGG CCCGCTAACA GCGCGATTTG  
4751  CTGGTGACCC AATGCGACCA GATGCTCCAC GCCCAGTCGC GTACCGTCTT  
4801  CATGGGAGAA AATAATACTG TTGATGGGTG TCTGGTCAGA GACATCAAGA  
4851  AATAACGCCG GAACATTAGT GCAGGCAGCT TCCACAGCAA TGGCATCCTG  
4901  GTCATCCAGC GGATAGTTAA TGATCAGCCC ACTGACGCGT TGCGCGAGAA  
4951  GATTGTGCAC CGCCGCTTTA CAGGCTTCGA CGCCGCTTCG TTCTACCATC  
5001  GACACCACCA CGCTGGCACC CAGTTGATCG GCGCGAGATT TAATCGCCGC  
5051  GACAATTTGC GACGGCGCGT GCAGGGCCAG ACTGGAGGTG GCAACGCCAA  
5101  TCAGCAACGA CTGTTTGCCC GCCAGTTGTT GTGCCACGCG GTTGGGAATG  
5151  TAATTCAGCT CCGCCATCGC CGCTTCCACT TTTTCCCGCG TTTTCGCAGA  
5201  AACGTGGCTG GCCTGGTTCA CCACGCGGGA AACGGTCTGA TAAGAGACAC  
5251  CGGCATACTC TGCGACATCG TATAACGTTA CTGGTTTCAC ATTCACCACC  
5301  CTGAATTGAC TCTCTTCCGG GCGCTATCAT GCCATACCGC GAAAGGTTTT  
5351  GCGCCATTCG ATGGTGTCCG GGATCTCGAC GCTCTCCCTT ATGCGACTCC  
5401  TGCATTAGGA AGCAGCCCAG TAGTAGGTTG AGGCCGTTGA GCACCGCCGC  
5451  CGCAAGGAAT GGTGCATGCA AGGAGATGGC GCCCAACAGT CCCCCGGCCA  
5501  CGGGGCCTGC CACCATACCC ACGCCGAAAC AAGCGCTCAT GAGCCCGAAG  
5551  TGGCGAGCCC GATCTTCCCC ATCGGTGATG TCGGCGATAT AGGCGCCAGC  
5601  AACCGCACCT GTGGCGCCGG TGATGCCGGC CACGATGCGT CCGGCGTAGA  
5651  GGATCGAGAT CTCGATCCCG CGAAAT