pcDNA6/TR

pcDNA6/TR

pcDNA6/TR载体基本信息

pcDNA6/TR载体质粒图谱和多克隆位点信息

pcDNA6/TR载体简介

T-REx四环素诱导系统

A Tetracycline-Regulated Expression System without Viral Transactivators

The T-REx System yields higher levels of induced expression than any other regulated mammalian expression system. It utilizes the complete CMV promoter and adds control elements

from the bacterial tetracycline resistance operon to effectively repress and derepress transcription from one of the strongest mammalian promoter sequences known (1,2).

Specific Activation

The T-REx System uses a repressor mechanism that blocks transcription from the powerful CMV promoter in the absence of tetracycline. Because the T-REx System elements do not use

viral transactivators, you can achieve high-level expression from the complete CMV promoter without secondary, non-specific activation of host genes.

The T-REx Mechanism

The T-REx transcriptional control elements are illustrated in Figure 1. Two tetracycline operator sequences (TetO2) have been inserted between the TATA box of the CMV promoter and the transcriptional start site. The TetO2 sequence itself has no effect on expression. When the tetracycline repressor protein (TR) is present, it effectively binds the TetO2 sites and blocks transcription initiation. Tetracycline added to the culture medium binds to, and changes the conformation of, the TR protein. This change causes the TR protein to release the TetO2 sites, derepressing transcription from the CMV promoter. The result is high-level expression of the gene of interest (Figure 2). Expression levels can be modulated based on the tetracycline concentration and can be induced to levels that are achieved with constitutive CMV expression vectors.

T-REx is a powerful inducible mammalian expression system that allows you to regulate expression from the complete human cytomegalovirus (CMV) enhancer-promoter. T-RExinducible expression vectors offer the following features:

 Complete CMV enhancer-promoter sequence containing two copies of the tetracycline operator TetO2 sequence for high-level regulated expression

 Zeocin or hygromycin resistance gene for effective selection of stable mammalian cell lines

 Large multiple cloning site to simplify cloning

In addition, pcDNA4/TO/myc-His offers a c-myc epitope for rapid detection of the recombinant protein with an Anti-myc Antibody and a polyhistidine (6xHis) sequence for simple purification of the recombinant protein with nickel-chelating resin and detection with Anti- His(C-term) Antibody.

The regulatory vector, pcDNA6/TR, is provided for high-level expression of the tetracycline repressor (TR) protein. This vector expresses the Blasticidin resistance gene for rapid selection of mammalian cell lines that stably express the TR protein.

pcDNA6/TR载体序列

ORIGIN

    1 GACGGATCGG GAGATCTCCC GATCCCCTAT GGTGCACTCT CAGTACAATC TGCTCTGATG

   61 CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCG

  121 CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGC

  181 TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATT

  241 GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATA

  301 TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACC

  361 CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCC

  421 ATTGACGTCA ATGGGTGGAG TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGT

  481 ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATT

  541 ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCA

  601 TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTG

  661 ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACC

  721 AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCG

  781 GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCA

  841 CTGCTTACTG GCTTATCGAA ATTAATACGA CTCACTATAG GGAGACCCAA GCTGGCTAGC

  901 GTTTAAACTT AAGCTTGGTA CCCGGGGATC CTCTAGGGCC TCTGAGCTAT TCCAGAAGTA

  961 GTGAAGAGGC TTTTTTGGAG GCCTAGGCTT TTGCAAAAAG CTCCGGATCG ATCCTGAGAA

 1021 CTTCAGGGTG AGTTTGGGGA CCCTTGATTG TTCTTTCTTT TTCGCTATTG TAAAATTCAT

 1081 GTTATATGGA GGGGGCAAAG TTTTCAGGGT GTTGTTTAGA ATGGGAAGAT GTCCCTTGTA

 1141 TCACCATGGA CCCTCATGAT AATTTTGTTT CTTTCACTTT CTACTCTGTT GACAACCATT

 1201 GTCTCCTCTT ATTTTCTTTT CATTTTCTGT AACTTTTTCG TTAAACTTTA GCTTGCATTT

 1261 GTAACGAATT TTTAAATTCA CTTTTGTTTA TTTGTCAGAT TGTAAGTACT TTCTCTAATC

 1321 ACTTTTTTTT CAAGGCAATC AGGGTATATT ATATTGTACT TCAGCACAGT TTTAGAGAAC

 1381 AATTGTTATA ATTAAATGAT AAGGTAGAAT ATTTCTGCAT ATAAATTCTG GCTGGCGTGG

 1441 AAATATTCTT ATTGGTAGAA ACAACTACAT CCTGGTCATC ATCCTGCCTT TCTCTTTATG

 1501 GTTACAATGA TATACACTGT TTGAGATGAG GATAAAATAC TCTGAGTCCA AACCGGGCCC

 1561 CTCTGCTAAC CATGTTCATG CCTTCTTCTT TTTCCTACAG CTCCTGGGCA ACGTGCTGGT

 1621 TATTGTGCTG TCTCATCATT TTGGCAAAGA ATTGTAATAC GACTCACTAT AGGGCGAATT

 1681 GATATGTCTA GATTAGATAA AAGTAAAGTG ATTAACAGCG CATTAGAGCT GCTTAATGAG

 1741 GTCGGAATCG AAGGTTTAAC AACCCGTAAA CTCGCCCAGA AGCTAGGTGT AGAGCAGCCT

 1801 ACATTGTATT GGCATGTAAA AAATAAGCGG GCTTTGCTCG ACGCCTTAGC CATTGAGATG

 1861 TTAGATAGGC ACCATACTCA CTTTTGCCCT TTAGAAGGGG AAAGCTGGCA AGATTTTTTA

 1921 CGTAATAACG CTAAAAGTTT TAGATGTGCT TTACTAAGTC ATCGCGATGG AGCAAAAGTA

 1981 CATTTAGGTA CACGGCCTAC AGAAAAACAG TATGAAACTC TCGAAAATCA ATTAGCCTTT

 2041 TTATGCCAAC AAGGTTTTTC ACTAGAGAAT GCATTATATG CACTCAGCGC TGTGGGGCAT

 2101 TTTACTTTAG GTTGCGTATT GGAAGATCAA GAGCATCAAG TCGCTAAAGA AGAAAGGGAA

 2161 ACACCTACTA CTGATAGTAT GCCGCCATTA TTACGACAAG CTATCGAATT ATTTGATCAC

 2221 CAAGGTGCAG AGCCAGCCTT CTTATTCGGC CTTGAATTGA TCATATGCGG ATTAGAAAAA

 2281 CAACTTAAAT GTGAAAGTGG GTCCGCGTAC AGCGGATCCC GGGAATTCAG ATCTTATTAA

 2341 AGCAGAACTT GTTTATTGCA GCTTATAATG GTTACAAATA AAGCAATAGC ATCACAAATT

 2401 TCACAAATAA AGCATTTTTT TCACTGCATT CTAGTTGTGG TTTGTCCAAA CTCATCAATG

 2461 TATCTTATCA TGTCTGGTCG AATCCATCAC ACTGGCGGCC GCTCGAGTCT AGAGGGCCCG

 2521 CGGTTCGAAG GTAAGCCTAT CCCTAACCCT CTCCTCGGTC TCGATTCTAC GCGTACCGGT

 2581 CATCATCACC ATCACCATTG AGTTTAAACC CGCTGATCAG CCTCGACTGT GCCTTCTAGT

 2641 TGCCAGCCAT CTGTTGTTTG CCCCTCCCCC GTGCCTTCCT TGACCCTGGA AGGTGCCACT

 2701 CCCACTGTCC TTTCCTAATA AAATGAGGAA ATTGCATCGC ATTGTCTGAG TAGGTGTCAT

 2761 TCTATTCTGG GGGGTGGGGT GGGGCAGGAC AGCAAGGGGG AGGATTGGGA AGACAATAGC

 2821 AGGCATGCTG GGGATGCGGT GGGCTCTATG GCTTCTGAGG CGGAAAGAAC CAGCTGGGGC

 2881 TCTAGGGGGT ATCCCCACGC GCCCTGTAGC GGCGCATTAA GCGCGGCGGG TGTGGTGGTT

 2941 ACGCGCAGCG TGACCGCTAC ACTTGCCAGC GCCCTAGCGC CCGCTCCTTT CGCTTTCTTC

 3001 CCTTCCTTTC TCGCCACGTT CGCCGGCTTT CCCCGTCAAG CTCTAAATCG GGGGCTCCCT

 3061 TTAGGGTTCC GATTTAGTGC TTTACGGCAC CTCGACCCCA AAAAACTTGA TTAGGGTGAT

 3121 GGTTCACGTA GTGGGCCATC GCCCTGATAG ACGGTTTTTC GCCCTTTGAC GTTGGAGTCC

 3181 ACGTTCTTTA ATAGTGGACT CTTGTTCCAA ACTGGAACAA CACTCAACCC TATCTCGGTC

 3241 TATTCTTTTG ATTTATAAGG GATTTTGCCG ATTTCGGCCT ATTGGTTAAA AAATGAGCTG

 3301 ATTTAACAAA AATTTAACGC GAATTAATTC TGTGGAATGT GTGTCAGTTA GGGTGTGGAA

 3361 AGTCCCCAGG CTCCCCAGCA GGCAGAAGTA TGCAAAGCAT GCATCTCAAT TAGTCAGCAA

 3421 CCAGGTGTGG AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC ATGCATCTCA

 3481 ATTAGTCAGC AACCATAGTC CCGCCCCTAA CTCCGCCCAT CCCGCCCCTA ACTCCGCCCA

 3541 GTTCCGCCCA TTCTCCGCCC CATGGCTGAC TAATTTTTTT TATTTATGCA GAGGCCGAGG

 3601 CCGCCTCTGC CTCTGAGCTA TTCCAGAAGT AGTGAGGAGG CTTTTTTGGA GGCCTAGGCT

 3661 TTTGCAAAAA GCTCCCGGGA GCTTGTATAT CCATTTTCGG ATCTGATCAG CACGTGTTGA

 3721 CAATTAATCA TCGGCATAGT ATATCGGCAT AGTATAATAC GACAAGGTGA GGAACTAAAC

 3781 CATGGCCAAG CCTTTGTCTC AAGAAGAATC CACCCTCATT GAAAGAGCAA CGGCTACAAT

 3841 CAACAGCATC CCCATCTCTG AAGACTACAG CGTCGCCAGC GCAGCTCTCT CTAGCGACGG

 3901 CCGCATCTTC ACTGGTGTCA ATGTATATCA TTTTACTGGG GGACCTTGTG CAGAACTCGT

 3961 GGTGCTGGGC ACTGCTGCTG CTGCGGCAGC TGGCAACCTG ACTTGTATCG TCGCGATCGG

 4021 AAATGAGAAC AGGGGCATCT TGAGCCCCTG CGGACGGTGC CGACAGGTGC TTCTCGATCT

 4081 GCATCCTGGG ATCAAAGCCA TAGTGAAGGA CAGTGATGGA CAGCCGACGG CAGTTGGGAT

 4141 TCGTGAATTG CTGCCCTCTG GTTATGTGTG GGAGGGCTAA GCACTTCGTG GCCGAGGAGC

 4201 AGGACTGACA CGTGCTACGA GATTTCGATT CCACCGCCGC CTTCTATGAA AGGTTGGGCT

 4261 TCGGAATCGT TTTCCGGGAC GCCGGCTGGA TGATCCTCCA GCGCGGGGAT CTCATGCTGG

 4321 AGTTCTTCGC CCACCCCAAC TTGTTTATTG CAGCTTATAA TGGTTACAAA TAAAGCAATA

 4381 GCATCACAAA TTTCACAAAT AAAGCATTTT TTTCACTGCA TTCTAGTTGT GGTTTGTCCA

 4441 AACTCATCAA TGTATCTTAT CATGTCTGTA TACCGTCGAC CTCTAGCTAG AGCTTGGCGT

 4501 AATCATGGTC ATAGCTGTTT CCTGTGTGAA ATTGTTATCC GCTCACAATT CCACACAACA

 4561 TACGAGCCGG AAGCATAAAG TGTAAAGCCT GGGGTGCCTA ATGAGTGAGC TAACTCACAT

 4621 TAATTGCGTT GCGCTCACTG CCCGCTTTCC AGTCGGGAAA CCTGTCGTGC CAGCTGCATT

 4681 AATGAATCGG CCAACGCGCG GGGAGAGGCG GTTTGCGTAT TGGGCGCTCT TCCGCTTCCT

 4741 CGCTCACTGA CTCGCTGCGC TCGGTCGTTC GGCTGCGGCG AGCGGTATCA GCTCACTCAA

 4801 AGGCGGTAAT ACGGTTATCC ACAGAATCAG GGGATAACGC AGGAAAGAAC ATGTGAGCAA

 4861 AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT GCTGGCGTTT TTCCATAGGC

 4921 TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG CGAAACCCGA

 4981 CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC

 5041 CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT

 5101 CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT

 5161 GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC TATCGTCTTG

 5221 AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT AACAGGATTA

 5281 GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT AACTACGGCT

 5341 ACACTAGAAG AACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC TTCGGAAAAA

 5401 GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTTTT TTTGTTTGCA

 5461 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC TTTTCTACGG

 5521 GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA

 5581 AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA

 5641 TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG

 5701 CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA

 5761 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC CCACGCTCAC

 5821 CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC

 5881 CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA

 5941 GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC GTGGTGTCAC

 6001 GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT

 6061 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC GTTGTCAGAA

 6121 GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC ACTGCATAAT TCTCTTACTG

 6181 TCATGCCATC CGTAAGATGC TTTTCTGTGA CTGGTGAGTA CTCAACCAAG TCATTCTGAG

 6241 AATAGTGTAT GCGGCGACCG AGTTGCTCTT GCCCGGCGTC AATACGGGAT AATACCGCGC

 6301 CACATAGCAG AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT

 6361 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA CCCAACTGAT

 6421 CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC AAAAACAGGA AGGCAAAATG

 6481 CCGCAAAAAA GGGAATAAGG GCGACACGGA AATGTTGAAT ACTCATACTC TTCCTTTTTC

 6541 AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA

 6601 TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG

 6661 TC

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