来源
2005-4-7 15:20:00
质粒pbgal-Basic信息
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| Restriction Map and Multiple Cloning Site of pbgal-Basic. Unique restriction sites are in bold. |
Note: The vector sequence file has been compiled from information in the sequence database, published literature, and other sources, together with partial sequences obtained by CLONTECH. This vector has not been completely sequenced.
Description
pbgal-Basic is a mammalian reporter vector designed for the analysis of the effects of cis-regulatory elements on gene expression in mammalian cells by assaying for beta-galactosidase. pbgal-Basic lacks eukaryotic promoter and enhancer sequences and has a polyadenylation signal upstream of the MCS to reduce background transcription; hence, it will not express significant levels of b-galactosidase. Therefore, the vector can serve as a negative control in many experiments and can also serve as a promoter-cloning vehicle for strong promoters. A pUC origin of replication and ampicillin resistance gene allow propagation and selection in E. coli. An f1 origin allows for single-stranded DNA production.
Feature Locations
- Multiple cloning site (MCS): 1–52
- b-galactosidase fusion protein
- Start codon (ATG): 94–96; stop codon: 3235–3237
Amino acids from D. melanogaster alcohol dehydrogenase: 94–186
Amino acids from E. coli b-galactosidase (1): 190-3234 - SV40 small t antigen intron: 3853-3918
- SV40 early mRNA polyadenylation signals: 4524–4529 & 4553–4558
- mRNA 3' ends: 4562 & 4574
- pUC plasmid origin of replication: 4908–5551
- Ampicillin resistance (b-lactamase) gene:
- Promoter: –35 region: 6629–6624; –10 region: 6606–6601
Transcription start point: 6594
Ribosome binding site: 6571–6567
b-lactamase coding sequences: start codon (ATG): 6559–6557; stop codon: 5701–5699
beta-lactamase signal peptide: 6559–6491; b-lactamase mature protein: 6490-5702 - f1 single-strand DNA origin (packages the coding strand of the lacZ gene): 6691–7146
- SV40 early mRNA polyadenylation signals: 7367–7372 & 7396–7401
全序列
CCCGGGAGGT ACCGAGCTCT TACGCGTGCT AGCTCGAGAT CTAAGTAAGC TTGGGGGATC GAAAGAGCCT GCTAAAGCAA AAAAGAAGTC ACCATGTCGT TTACTTTGAC CAACAAGAAC GTGATTTTCG TTGCCGGTCT GGGAGGCATT GGTCTGGACA CCAGCAAGGA GCTGCTCAAG CGCGATCCCG TCGTTTTACA ACGTCGTGAC TGGGAAAACC CTGGCGTTAC CCAACTTAAT CGCCTTGCAG CACATCCCCC TTTCGCCAGC TGGCGTAATA GCGAAGAGGC CCGCACCGAT CGCCCTTCCC AACAGTTGCG CAGCCTGAAT GGCGAATGGC GCTTTGCCTG GTTTCCGGCA CCAGAAGCGG TGCCGGAAAG CTGGCTGGAG TGCGATCTTC CTGAGGCCGA TACTGTCGTC GTCCCCTCAA ACTGGCAGAT GCACGGTTAC GATGCGCCCA TCTACACCAA CGTAACCTAT CCCATTACGG TCAATCCGCC GTTTGTTCCC ACGGAGAATC CGACGGGTTG TTACTCGCTC ACATTTAATG TTGATGAAAG CTGGCTACAG GAAGGCCAGA CGCGAATTAT TTTTGATGGC GTTAACTCGG CGTTTCATCT GTGGTGCAAC GGGCGCTGGG TCGGTTACGG CCAGGACAGT CGTTTGCCGT CTGAATTTGA CCTGAGCGCA TTTTTACGCG CCGGAGAAAA CCGCCTCGCG GTGATGGTGC TGCGTTGGAG TGACGGCAGT TATCTGGAAG ATCAGGATAT GTGGCGGATG AGCGGCATTT TCCGTGACGT CTCGTTGCTG CATAAACCGA CTACACAAAT CAGCGATTTC CATGTTGCCA CTCGCTTTAA TGATGATTTC AGCCGCGCTG TACTGGAGGC TGAAGTTCAG ATGTGCGGCG AGTTGCGTGA CTACCTACGG GTAACAGTTT CTTTATGGCA GGGTGAAACG CAGGTCGCCA GCGGCACCGC GCCTTTCGGC GGTGAAATTA TCGATGAGCG TGGTGGTTAT GCCGATCGCG TCACACTACG TCTGAACGTC GAAAACCCGA AACTGTGGAG CGCCGAAATC CCGAATCTCT ATCGTGCGGT GGTTGAACTG CACACCGCCG ACGGCACGCT GATTGAAGCA GAAGCCTGCG ATGTCGGTTT CCGCGAGGTG CGGATTGAAA ATGGTCTGCT GCTGCTGAAC GGCAAGCCGT TGCTGATTCG AGGCGTTAAC CGTCACGAGC ATCATCCTCT GCATGGTCAG GTCATGGATG AGCAGACGAT GGTGCAGGAT ATCCTGCTGA TGAAGCAGAA CAACTTTAAC GCCGTGCGCT GTTCGCATTA TCCGAACCAT CCGCTGTGGT ACACGCTGTG CGACCGCTAC GGCCTGTATG TGGTGGATGA AGCCAATATT GAAACCCACG GCATGGTGCC AATGAATCGT CTGACCGATG ATCCGCGCTG GCTACCGGCG ATGAGCGAAC GCGTAACGCG AATGGTGCAG CGCGATCGTA ATCACCCGAG TGTGATCATC TGGTCGCTGG GGAATGAATC AGGCCACGGC GCTAATCACG ACGCGCTGTA TCGCTGGATC AAATCTGTCG ATCCTTCCCG CCCGGTGCAG TATGAAGGCG GCGGAGCCGA CACCACGGCC ACCGATATTA TTTGCCCGAT GTACGCGCGC GTGGATGAAG ACCAGCCCTT CCCGGCTGTG CCGAAATGGT CCATCAAAAA ATGGCTTTCG CTACCTGGAG AGACGCGCCC GCTGATCCTT TGCGAATACG CCCACGCGAT GGGTAACAGT CTTGGCGGTT TCGCTAAATA CTGGCAGGCG TTTCGTCAGT ATCCCCGTTT ACAGGGCGGC TTCGTCTGGG ACTGGGTGGA TCAGTCGCTG ATTAAATATG ATGAAAACGG CAACCCGTGG TCGGCTTACG GCGGTGATTT TGGCGATACG CCGAACGATC GCCAGTTCTG TATGAACGGT CTGGTCTTTG CCGACCGCAC GCCGCATCCA GCGCTGACGG AAGCAAAACA CCAGCAGCAG TTTTTCCAGT TCCGTTTATC CGGGCAAACC ATCGAAGTGA CCAGCGAATA CCTGTTCCGT CATAGCGATA ACGAGCTCCT GCACTGGATG GTGGCGCTGG ATGGTAAGCC GCTGGCAAGC GGTGAAGTGC CTCTGGATGT CGCTCCACAA GGTAAACAGT TGATTGAACT GCCTGAACTA CCGCAGCCGG AGAGCGCCGG GCAACTCTGG CTCACAGTAC GCGTAGTGCA ACCGAACGCG ACCGCATGGT CAGAAGCCGG GCACATCAGC GCCTGGCAGC AGTGGCGTCT GGCGGAAAAC CTCAGTGTGA CGCTCCCCGC CGCGTCCCAC GCCATCCCGC ATCTGACCAC CAGCGAAATG GATTTTTGCA TCGAGCTGGG TAATAAGCGT TGGCAATTTA ACCGCCAGTC AGGCTTTCTT TCACAGATGT GGATTGGCGA TAAAAAACAA CTGCTGACGC CGCTGCGCGA TCAGTTCACC CGTGCACCGC TGGATAACGA CATTGGCGTA AGTGAAGCGA CCCGCATTGA CCCTAACGCC TGGGTCGAAC GCTGGAAGGC GGCGGGCCAT TACCAGGCCG AAGCAGCGTT GTTGCAGTGC ACGGCAGATA CACTTGCTGA TGCGGTGCTG ATTACGACCG CTCACGCGTG GCAGCATCAG GGGAAAACCT TATTTATCAG CCGGAAAACC TACCGGATTG ATGGTAGTGG TCAAATGGCG ATTACCGTTG ATGTTGAAGT GGCGAGCGAT ACACCGCATC CGGCGCGGAT TGGCCTGAAC TGCCAGCTGG CGCAGGTAGC AGAGCGGGTA AACTGGCTCG GATTAGGGCC GCAAGAAAAC TATCCCGACC GCCTTACTGC CGCCTGTTTT GACCGCTGGG ATCTGCCATT GTCAGACATG TATACCCCGT ACGTCTTCCC GAGCGAAAAC GGTCTGCGCT GCGGGACGCG CGAATTGAAT TATGGCCCAC ACCAGTGGCG CGGCGACTTC CAGTTCAACA TCAGCCGCTA CAGTCAACAG CAACTGATGG AAACCAGCCA TCGCCATCTG CTGCACGCGG AAGAAGGCAC ATGGCTGAAT ATCGACGGTT TCCATATGGG GATTGGTGGC GACGACTCCT GGAGCCCGTC AGTATCGGCG GAATTACAGC TGAGCGCCGG TCGCTACCAT TACCAGTTGG TCTGGTGTCA AAAATAATAA TAACCGGGCA GGCCATGTCT GCCCGTATTT CGCGTAAGGA AATCCATTAT GTACTATTTA TCAGGTGGCC CCCGCTGAAT TGGAATCGAT ATTGTTACAA CACCCCAACA TCTTCGACGC GGGCGTGGCA GGTCTTCCCG ACGATGACGC CGGTGAACTT CCCGCCGCCG TTGTTGTTTT GGAGCACGGA AAGACGATGA CGGAAAAAGA GATCGTGGAT TACGTCGCCA GTCAAGTAAC AACCGCGAAA AAGTTGCGCG GAGGAGTTGT GTTTGTGGAC GAAGTACCGA AAGGTCTTAC CGGAAAACTC GACGCAAGAA AAATCAGAGA GATCCTCATA AAGGCCAAGA AGGGCGGAAA GTCCAAATTG TAAAATGTAA CTGTATTCAG CGATGACGAA ATTCTTAGCT ATTGTAATAC TGCGATGAGT GGCAGGGCGG GGCGTAATTT TTTTAAGGCA GTTATTGGTG CCCTTAAACG CCTGGTGCTA CGCCTGAATA AGTGATAATA AGCGGATGAA TGGCAGAAAT TCGCCGGATC TTTGTGAAGG AACCTTACTT CTGTGGTGTG ACATAATTGG ACAAACTACC TACAGAGATT TAAAGCTCTA AGGTAAATAT AAAATTTTTA AGTGTATAAT GTGTTAAACT ACTGATTCTA ATTGTTTGTG TATTTTAGAT TCCAACCTAT GGAACTGATG AATGGGAGCA GTGGTGGAAT GCCTTTAATG AGGAAAACCT GTTTTGCTCA GAAGAAATGC CATCTAGTGA TGATGAGGCT ACTGCTGACT CTCAACATTC TACTCCTCCA AAAAAGAAGA GAAAGGTAGA AGACCCCAAG GACTTTCCTT CAGAATTGCT AAGTTTTTTG AGTCATGCTG TGTTTAGTAA TAGAACTCTT GCTTGCTTTG CTATTTACAC CACAAAGGAA AAAGCTGCAC TGCTATACAA GAAAATTATG GAAAAATATT CTGTAACCTT TATAAGTAGG CATAACAGTT ATAATCATAA CATACTGTTT TTTCTTACTC CACACAGGCA TAGAGTGTCT GCTATTAATA ACTATGCTCA AAAATTGTGT ACCTTTAGCT TTTTAATTTG TAAAGGGGTT AATAAGGAAT ATTTGATGTA TAGTGCCTTG ACTAGAGATC ATAATCAGCC ATACCACATT TGTAGAGGTT TTACTTGCTT TAAAAAACCT CCCACACCTC CCCCTGAACC TGAAACATAA AATGAATGCA ATTGTTGTTG TTAACTTGTT TATTGCAGCT TATAATGGTT ACAAATAAAG CAATAGCATC ACAAATTTCA CAAATAAAGC ATTTTTTTCA CTGCATTCTA GTTGTGGTTT GTCCAAACTC ATCAATGTAT CTTATCATGT CTGGATCCGT CGACCGATGC CCTTGAGAGC CTTCAACCCA GTCAGCTCCT TCCGGTGGGC GCGGGGCATG ACTATCGTCG CCGCACTTAT GACTGTCTTC TTTATCATGC AACTCGTAGG ACAGGTGCCG GCAGCGCTCT TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC GGCTGCGGCG AGCGGTATCA GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG GGGATAACGC AGGAAAGAAC ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA AGATCCTTTG ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA ATTAAAAATG AAGTTTTAAA TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT ACCAATGCTT AATCAGTGAG GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG TTGCCTGACT CCCCGTCGTG TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA GTGCTGCAAT GATACCGCGA GACCCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC AGCCAGCCGG AAGGGCCGAG CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT CTATTAATTG TTGCCGGGAA GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG TTGTTGCCAT TGCTACAGGC ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA GCTCCGGTTC CCAACGATCA AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG TTAGCTCCTT CGGTCCTCCG ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA TGGTTATGGC AGCACTGCAT AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG TGACTGGTGA GTACTCAACC AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT CTTGCCCGGC GTCAATACGG GATAATACCG CGCCACATAG CAGAACTTTA AAAGTGCTCA TCATTGGAAA ACGTTCTTCG GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA GTTCGATGTA ACCCACTCGT GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG TTTCTGGGTG AGCAAAAACA GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC GGAAATGTTG AATACTCATA CTCTTCCTTT TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC ATATTTGAAT GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGCACATT TCCCCGAAAA GTGCCACCTG ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG CGGGTGTGGT GGTTACGCGC AGCGTGACCG CTACACTTGC CAGCGCCCTA GCGCCCGCTC CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA ATCGGGGGCT CCCTTTAGGG TTCCGATTTA GTGCTTTACG GCACCTCGAC CCCAAAAAAC TTGATTAGGG TGATGGTTCA CGTAGTGGGC CATCGCCCTG ATAGACGGTT TTTCGCCCTT TGACGTTGGA GTCCACGTTC TTTAATAGTG GACTCTTGTT CCAAACTGGA ACAACACTCA ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGGGATTTT GCCGATTTCG GCCTATTGGT TAAAAAATGA GCTGATTTAA CAAAAATTTA ACGCGAATTT TAACAAAATA TTAACGTTTA CAATTTCCCA TTCGCCATTC AGGCTACGCA ACTGTTGGGA AGGGCGATCG GTGCGGGCCT CTTCGCTATT ACGCCAGCCC AAGCTACCAT GATAAGTAAG TAATATTAAG GTACGTGGAG GTTTTACTTG CTTTAAAAAA CCTCCCACAC CTCCCCCTGA ACCTGAAACA TAAAATGAAT GCAATTGTTG TTGTTAACTT GTTTATTGCA GCTTATAATG GTTACAAATA AAGCAATAGC ATCACAAATT TCACAAATAA AGCATTTTTT TCACTGCATT CTAGTTGTGG TTTGTCCAAA CTCATCAATG TATCTTATGG TACTGTAACT GAGCTAACAT AA
