质粒pbgalControl信息_实验方法_

Description

pbgal-Controlisareportervectorwhichwillexpressb-galactosidaseinmammaliancells.Therefore,thisvectorcanserveasapositivecontrolinexperimentsthatuselacZastheprimaryreporterandcanalsobeusedtonormalizetransfectionefficienciesinexperimentsusingotherreportermolecules.pbgal-ControlcontainstheSV40earlypromoterandenhancersequencesinsertedupstreamanddownstream,respectively,ofthelacZgene.ThepromoterfragmentcontainsanSV40originofreplicationwhichisactiveincellsexpressingtheSV40T-antigen.ApUCoriginofreplicationandampicillinresistancegeneallowpropagationandselectioninE.coli.Anf1originallowsforsingle-strandedDNAproduction.

FeatureLocations

• Multiplecloningsite(MCS):1–42
• SV40earlypromoter
  Promoterinsert:38–23921-bprepeats:69–89,90–110&112–132Earlypromoterelement:145–151;majortranscriptionstartpoints:141,179,185&190
• SV40originofreplication:89–224
• b-galactosidasefusionprotein
  Startcodon(ATG):286–288;stopcodon:3427–3429AminoacidsfromD.melanogasteralcoholdehydrogenase:286–378AminoacidsfromE.colib-galactosidase(1):382–3426
• SV40smalltantigenintron:4045–4110
• SV40earlymRNApolyadenylationsignals:4716–4721&4745–4750
  mRNA3ends:4754&4766
• SV40enhancer:
  Enhancerinsert:4816–507272-bptandemrepeats:4844–4915&4916–498721-bprepeats:4991–5011,5012–5032&5034–5054
• pUCplasmidoriginofreplication:4908–5551
• Ampicillinresistance(b-lactamase)gene:
  Promoter:–35region:6629–6624;–10region:6606–6601Transcriptionstartpoint:6594Ribosomebindingsite:6571–6567b-lactamasecodingsequences:startcodon(ATG):6559–6557;stopcodon:5701–5699b-lactamasesignalpeptide:6559–6491;b-lactamasematureprotein:6490–5702
• f1single-strandDNAorigin(packagesthecodingstrandofthelacZgene):6691–7146
• SV40earlymRNApolyadenylationsignals:7367–7372&7396–7401

全序列

CCCGGGAGGT ACCGAGCTCT TACGCGTGCT AGCTCGAGAT CTGCATCTCA ATTAGTCAGCAACCATAGTC CCGCCCCTAA CTCCGCCCAT CCCGCCCCTA ACTCCGCCCA GTTCCGCCCATTCTCCGCCC CATGGCTGAC TAATTTTTTT TATTTATGCA GAGGCCGAGG CCGCCTCGGCCTCTGAGCTA TTCCAGAAGT AGTGAGGAGG CTTTTTTGGA GGCCTAGGCT TTTGCAAAAAGCTTGGGGGA TCGAAAGAGC CTGCTAAAGC AAAAAAGAAG TCACCATGTC GTTTACTTTGACCAACAAGA ACGTGATTTT CGTTGCCGGT CTGGGAGGCA TTGGTCTGGA CACCAGCAAGGAGCTGCTCA AGCGCGATCC CGTCGTTTTA CAACGTCGTG ACTGGGAAAA CCCTGGCGTTACCCAACTTA ATCGCCTTGC AGCACATCCC CCTTTCGCCA GCTGGCGTAA TAGCGAAGAGGCCCGCACCG ATCGCCCTTC CCAACAGTTG CGCAGCCTGA ATGGCGAATG GCGCTTTGCCTGGTTTCCGG CACCAGAAGC GGTGCCGGAA AGCTGGCTGG AGTGCGATCT TCCTGAGGCCGATACTGTCG TCGTCCCCTC AAACTGGCAG ATGCACGGTT ACGATGCGCC CATCTACACCAACGTAACCT ATCCCATTAC GGTCAATCCG CCGTTTGTTC CCACGGAGAA TCCGACGGGTTGTTACTCGC TCACATTTAA TGTTGATGAA AGCTGGCTAC AGGAAGGCCA GACGCGAATTATTTTTGATG GCGTTAACTC GGCGTTTCAT CTGTGGTGCA ACGGGCGCTG GGTCGGTTACGGCCAGGACA GTCGTTTGCC GTCTGAATTT GACCTGAGCG CATTTTTACG CGCCGGAGAAAACCGCCTCG CGGTGATGGT GCTGCGTTGG AGTGACGGCA GTTATCTGGA AGATCAGGATATGTGGCGGA TGAGCGGCAT TTTCCGTGAC GTCTCGTTGC TGCATAAACC GACTACACAAATCAGCGATT TCCATGTTGC CACTCGCTTT AATGATGATT TCAGCCGCGC TGTACTGGAGGCTGAAGTTC AGATGTGCGG CGAGTTGCGT GACTACCTAC GGGTAACAGT TTCTTTATGGCAGGGTGAAA CGCAGGTCGC CAGCGGCACC GCGCCTTTCG GCGGTGAAAT TATCGATGAGCGTGGTGGTT ATGCCGATCG CGTCACACTA CGTCTGAACG TCGAAAACCC GAAACTGTGGAGCGCCGAAA TCCCGAATCT CTATCGTGCG GTGGTTGAAC TGCACACCGC CGACGGCACGCTGATTGAAG CAGAAGCCTG CGATGTCGGT TTCCGCGAGG TGCGGATTGA AAATGGTCTGCTGCTGCTGA ACGGCAAGCC GTTGCTGATT CGAGGCGTTA ACCGTCACGA GCATCATCCTCTGCATGGTC AGGTCATGGA TGAGCAGACG ATGGTGCAGG ATATCCTGCT GATGAAGCAGAACAACTTTA ACGCCGTGCG CTGTTCGCAT TATCCGAACC ATCCGCTGTG GTACACGCTGTGCGACCGCT ACGGCCTGTA TGTGGTGGAT GAAGCCAATA TTGAAACCCA CGGCATGGTGCCAATGAATC GTCTGACCGA TGATCCGCGC TGGCTACCGG CGATGAGCGA ACGCGTAACGCGAATGGTGC AGCGCGATCG TAATCACCCG AGTGTGATCA TCTGGTCGCT GGGGAATGAATCAGGCCACG GCGCTAATCA CGACGCGCTG TATCGCTGGA TCAAATCTGT CGATCCTTCCCGCCCGGTGC AGTATGAAGG CGGCGGAGCC GACACCACGG CCACCGATAT TATTTGCCCGATGTACGCGC GCGTGGATGA AGACCAGCCC TTCCCGGCTG TGCCGAAATG GTCCATCAAAAAATGGCTTT CGCTACCTGG AGAGACGCGC CCGCTGATCC TTTGCGAATA CGCCCACGCGATGGGTAACA GTCTTGGCGG TTTCGCTAAA TACTGGCAGG CGTTTCGTCA GTATCCCCGTTTACAGGGCG GCTTCGTCTG GGACTGGGTG GATCAGTCGC TGATTAAATA TGATGAAAACGGCAACCCGT GGTCGGCTTA CGGCGGTGAT TTTGGCGATA CGCCGAACGA TCGCCAGTTCTGTATGAACG GTCTGGTCTT TGCCGACCGC ACGCCGCATC CAGCGCTGAC GGAAGCAAAACACCAGCAGC AGTTTTTCCA GTTCCGTTTA TCCGGGCAAA CCATCGAAGT GACCAGCGAATACCTGTTCC GTCATAGCGA TAACGAGCTC CTGCACTGGA TGGTGGCGCT GGATGGTAAGCCGCTGGCAA GCGGTGAAGT GCCTCTGGAT GTCGCTCCAC AAGGTAAACA GTTGATTGAACTGCCTGAAC TACCGCAGCC GGAGAGCGCC GGGCAACTCT GGCTCACAGT ACGCGTAGTGCAACCGAACG CGACCGCATG GTCAGAAGCC GGGCACATCA GCGCCTGGCA GCAGTGGCGTCTGGCGGAAA ACCTCAGTGT GACGCTCCCC GCCGCGTCCC ACGCCATCCC GCATCTGACCACCAGCGAAA TGGATTTTTG CATCGAGCTG GGTAATAAGC GTTGGCAATT TAACCGCCAGTCAGGCTTTC TTTCACAGAT GTGGATTGGC GATAAAAAAC AACTGCTGAC GCCGCTGCGCGATCAGTTCA CCCGTGCACC GCTGGATAAC GACATTGGCG TAAGTGAAGC GACCCGCATTGACCCTAACG CCTGGGTCGA ACGCTGGAAG GCGGCGGGCC ATTACCAGGC CGAAGCAGCGTTGTTGCAGT GCACGGCAGA TACACTTGCT GATGCGGTGC TGATTACGAC CGCTCACGCGTGGCAGCATC AGGGGAAAAC CTTATTTATC AGCCGGAAAA CCTACCGGAT TGATGGTAGTGGTCAAATGG CGATTACCGT TGATGTTGAA GTGGCGAGCG ATACACCGCA TCCGGCGCGGATTGGCCTGA ACTGCCAGCT GGCGCAGGTA GCAGAGCGGG TAAACTGGCT CGGATTAGGGCCGCAAGAAA ACTATCCCGA CCGCCTTACT GCCGCCTGTT TTGACCGCTG GGATCTGCCATTGTCAGACA TGTATACCCC GTACGTCTTC CCGAGCGAAA ACGGTCTGCG CTGCGGGACGCGCGAATTGA ATTATGGCCC ACACCAGTGG CGCGGCGACT TCCAGTTCAA CATCAGCCGCTACAGTCAAC AGCAACTGAT GGAAACCAGC CATCGCCATC TGCTGCACGC GGAAGAAGGCACATGGCTGA ATATCGACGG TTTCCATATG GGGATTGGTG GCGACGACTC CTGGAGCCCGTCAGTATCGG CGGAATTACA GCTGAGCGCC GGTCGCTACC ATTACCAGTT GGTCTGGTGTCAAAAATAAT AATAACCGGG CAGGCCATGT CTGCCCGTAT TTCGCGTAAG GAAATCCATTATGTACTATT TATCAGGTGG CCCCCGCTGA ATTGGAATCG ATATTGTTAC AACACCCCAACATCTTCGAC GCGGGCGTGG CAGGTCTTCC CGACGATGAC GCCGGTGAAC TTCCCGCCGCCGTTGTTGTT TTGGAGCACG GAAAGACGAT GACGGAAAAA GAGATCGTGG ATTACGTCGCCAGTCAAGTA ACAACCGCGA AAAAGTTGCG CGGAGGAGTT GTGTTTGTGG ACGAAGTACCGAAAGGTCTT ACCGGAAAAC TCGACGCAAG AAAAATCAGA GAGATCCTCA TAAAGGCCAAGAAGGGCGGA AAGTCCAAAT TGTAAAATGT AACTGTATTC AGCGATGACG AAATTCTTAGCTATTGTAAT ACTGCGATGA GTGGCAGGGC GGGGCGTAAT TTTTTTAAGG CAGTTATTGGTGCCCTTAAA CGCCTGGTGC TACGCCTGAA TAAGTGATAA TAAGCGGATG AATGGCAGAAATTCGCCGGA TCTTTGTGAA GGAACCTTAC TTCTGTGGTG TGACATAATT GGACAAACTACCTACAGAGA TTTAAAGCTC TAAGGTAAAT ATAAAATTTT TAAGTGTATA ATGTGTTAAACTACTGATTC TAATTGTTTG TGTATTTTAG ATTCCAACCT ATGGAACTGA TGAATGGGAGCAGTGGTGGA ATGCCTTTAA TGAGGAAAAC CTGTTTTGCT CAGAAGAAAT GCCATCTAGTGATGATGAGG CTACTGCTGA CTCTCAACAT TCTACTCCTC CAAAAAAGAA GAGAAAGGTAGAAGACCCCA AGGACTTTCC TTCAGAATTG CTAAGTTTTT TGAGTCATGC TGTGTTTAGTAATAGAACTC TTGCTTGCTT TGCTATTTAC ACCACAAAGG AAAAAGCTGC ACTGCTATACAAGAAAATTA TGGAAAAATA TTCTGTAACC TTTATAAGTA GGCATAACAG TTATAATCATAACATACTGT TTTTTCTTAC TCCACACAGG CATAGAGTGT CTGCTATTAA TAACTATGCTCAAAAATTGT GTACCTTTAG CTTTTTAATT TGTAAAGGGG TTAATAAGGA ATATTTGATGTATAGTGCCT TGACTAGAGA TCATAATCAG CCATACCACA TTTGTAGAGG TTTTACTTGCTTTAAAAAAC CTCCCACACC TCCCCCTGAA CCTGAAACAT AAAATGAATG CAATTGTTGTTGTTAACTTG TTTATTGCAG CTTATAATGG TTACAAATAA AGCAATAGCA TCACAAATTTCACAAATAAA GCATTTTTTT CACTGCATTC TAGTTGTGGT TTGTCCAAAC TCATCAATGTATCTTATCAT GTCTGGATCT GCTGTGGAAT GTGTGTCAGT TAGGGTGTGG AAAGTCCCCAGGCTCCCCAG CAGGCAGAAG TATGCAAAGC ATGCATCTCA ATTAGTCAGC AACCAGGTGTGGAAAGTCCC CAGGCTCCCC AGCAGGCAGA AGTATGCAAA GCATGCATCT CAATTAGTCAGCAACCATAG TCCCGCCCCT AACTCCGCCC ATCCCGCCCC TAACTCCGCC CAGTTCCGCCCATTCTCCGC CCCATGGTTC AGATCCTCTA GGGATCCGTC GACCGATGCC CTTGAGAGCCTTCAACCCAG TCAGCTCCTT CCGGTGGGCG CGGGGCATGA CTATCGTCGC CGCACTTATGACTGTCTTCT TTATCATGCA ACTCGTAGGA CAGGTGCCGG CAGCGCTCTT CCGCTTCCTCGCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG CTCACTCAAAGGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA TGTGAGCAAAAGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT TCCATAGGCTCCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC GAAACCCGACAGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT CTCCTGTTCCGACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG TGGCGCTTTCTCATAGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA AGCTGGGCTGTGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT ATCGTCTTGAGTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA ACAGGATTAGCAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA ACTACGGCTACACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT TCGGAAAAAGAGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT TTTTTGTTTGCAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA TCTTTTCTACGGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA TGAGATTATCAAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT CAATCTAAAGTATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG CACCTATCTCAGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT AGATAACTACGATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG ACCCACGCTCACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC GCAGAAGTGGTCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG CTAGAGTAAGTAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA TCGTGGTGTCACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA GGCGAGTTACATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA TCGTTGTCAGAAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA ATTCTCTTACTGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA AGTCATTCTGAGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG ATAATACCGCGCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG GGCGAAAACTCTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG CACCCAACTGATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG GAAGGCAAAATGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC TCTTCCTTTTTCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATGTATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG TGCCACCTGACGCGCCCTGT AGCGGCGCAT TAAGCGCGGC GGGTGTGGTG GTTACGCGCA GCGTGACCGCTACACTTGCC AGCGCCCTAG CGCCCGCTCC TTTCGCTTTC TTCCCTTCCT TTCTCGCCACGTTCGCCGGC TTTCCCCGTC AAGCTCTAAA TCGGGGGCTC CCTTTAGGGT TCCGATTTAGTGCTTTACGG CACCTCGACC CCAAAAAACT TGATTAGGGT GATGGTTCAC GTAGTGGGCCATCGCCCTGA TAGACGGTTT TTCGCCCTTT GACGTTGGAG TCCACGTTCT TTAATAGTGGACTCTTGTTC CAAACTGGAA CAACACTCAA CCCTATCTCG GTCTATTCTT TTGATTTATAAGGGATTTTG CCGATTTCGG CCTATTGGTT AAAAAATGAG CTGATTTAAC AAAAATTTAACGCGAATTTT AACAAAATAT TAACGTTTAC AATTTCCCAT TCGCCATTCA GGCTACGCAACTGTTGGGAA GGGCGATCGG TGCGGGCCTC TTCGCTATTA CGCCAGCCCA AGCTACCATGATAAGTAAGT AATATTAAGG TACGTGGAGG TTTTACTTGC TTTAAAAAAC CTCCCACACCTCCCCCTGAA CCTGAAACAT AAAATGAATG CAATTGTTGT TGTTAACTTG TTTATTGCAGCTTATAATGG TTACAAATAA AGCAATAGCA TCACAAATTT CACAAATAAA GCATTTTTTTCACTGCATTC TAGTTGTGGT TTGTCCAAAC TCATCAATGT ATCTTATGGT ACTGTAACTGAGCTAACATA A