pTAL-Luc

pTAL-Luc is designed for analyzing enhancer sequences by assaying for expression of the firefly luciferase (luc) gene from Photinus pyralis (1). This vector contains a TATA-like promoter (PTAL) region from the Herpes simplex virus thymidine kinase (HSV-TK) promoter. Putative enhancers can be inserted into one of the MCSs. The luciferase coding sequence is followed by the SV40 late polyadenylation signal to ensure proper, efficient processing of the luc transcript in eukaryotic cells. Located upstream of the MCS is a synthetic transcription blocker (TB), which is composed of adjacent polyadenylation and transcription pause sites for reducing background transcription (2). The vector backbone also contains an f1 origin for single-stranded DNA production, a pUC origin of replication, and an ampicillin resistance gene for propagation and selection in E. coli.

载体应用

pTAL-Luc is ideal for use as a negative control or for studying putative enhancers that are inserted upstream of the luciferase reporter gene. Luciferase is a highly sensitive enzymatic reporter that can be assayed by any standard luciferase-detection method, providing quantitative data on induction levels. The pTAL-Luc Vector can be transfected into mammalian cells by any standard method. For selecting stable clones, cotransfect with a vector containing an antibiotic resistance gene, such as neomycin, hygromycin or puromycin.

0001 GGTACCGAGC TCTTACGCGT GCTAGCCCGG GCTCGAGATC TGCCGCCCCG ACTGCATCTG 
0061 CGTGTTCGAA TTCGCCAATG ACAAGACGCT GGGCGGGGTT TGTGTCATCA TAGAACTAAA 
0121 GACATGCAAA TATATTTCTT CCGGGGACAC CGCCAGCAAA CGCGAGCAAC GGGCCACGGG 
0181 GATGAAGCAG AAGCTTGGCA TTCCGGTACT GTTGGTAAAG CCACCATGGA AGACGCCAAA 
0241 AACATAAAGA AAGGCCCGGC GCCATTCTAT CCGCTGGAAG ATGGAACCGC TGGAGAGCAA 
0301 CTGCATAAGG CTATGAAGAG ATACGCCCTG GTTCCTGGAA CAATTGCTTT TACAGATGCA 
0361 CATATCGAGG TGGACATCAC TTACGCTGAG TACTTCGAAA TGTCCGTTCG GTTGGCAGAA 
0421 GCTATGAAAC GATATGGGCT GAATACAAAT CACAGAATCG TCGTATGCAG TGAAAACTCT 
0481 CTTCAATTCT TTATGCCGGT GTTGGGCGCG TTATTTATCG GAGTTGCAGT TGCGCCCGCG 
0541 AACGACATTT ATAATGAACG TGAATTGCTC AACAGTATGG GCATTTCGCA GCCTACCGTG 
0601 GTGTTCGTTT CCAAAAAGGG GTTGCAAAAA ATTTTGAACG TGCAAAAAAA GCTCCCAATC 
0661 ATCCAAAAAA TTATTATCAT GGATTCTAAA ACGGATTACC AGGGATTTCA GTCGATGTAC 
0721 ACGTTCGTCA CATCTCATCT ACCTCCCGGT TTTAATGAAT ACGATTTTGT GCCAGAGTCC 
0781 TTCGATAGGG ACAAGACAAT TGCACTGATC ATGAACTCCT CTGGATCTAC TGGTCTGCCT 
0841 AAAGGTGTCG CTCTGCCTCA TAGAACTGCC TGCGTGAGAT TCTCGCATGC CAGAGATCCT 
0901 ATTTTTGGCA ATCAAATCAT TCCGGATACT GCGATTTTAA GTGTTGTTCC ATTCCATCAC 
0961 GGTTTTGGAA TGTTTACTAC ACTCGGATAT TTGATATGTG GATTTCGAGT CGTCTTAATG 
1021 TATAGATTTG AAGAAGAGCT GTTTCTGAGG AGCCTTCAGG ATTACAAGAT TCAAAGTGCG 
1081 CTGCTGGTGC CAACCCTATT CTCCTTCTTC GCCAAAAGCA CTCTGATTGA CAAATACGAT 
1141 TTATCTAATT TACACGAAAT TGCTTCTGGT GGCGCTCCCC TCTCTAAGGA AGTCGGGGAA 
1201 GCGGTTGCCA AGAGGTTCCA TCTGCCAGGT ATCAGGCAAG GATATGGGCT CACTGAGACT 
1261 ACATCAGCTA TTCTGATTAC ACCCGAGGGG GATGATAAAC CGGGCGCGGT CGGTAAAGTT 
1321 GTTCCATTTT TTGAAGCGAA GGTTGTGGAT CTGGATACCG GGAAAACGCT GGGCGTTAAT 
1381 CAAAGAGGCG AACTGTGTGT GAGAGGTCCT ATGATTATGT CCGGTTATGT AAACAATCCG 
1441 GAAGCGACCA ACGCCTTGAT TGACAAGGAT GGATGGCTAC ATTCTGGAGA CATAGCTTAC 
1501 TGGGACGAAG ACGAACACTT CTTCATCGTT GACCGCCTGA AGTCTCTGAT TAAGTACAAA 
1561 GGCTATCAGG TGGCTCCCGC TGAATTGGAA TCCATCTTGC TCCAACACCC CAACATCTTC 
1621 GACGCAGGTG TCGCAGGTCT TCCCGACGAT GACGCCGGTG AACTTCCCGC CGCCGTTGTT 
1681 GTTTTGGAGC ACGGAAAGAC GATGACGGAA AAAGAGATCG TGGATTACGT CGCCAGTCAA 
1741 GTAACAACCG CGAAAAAGTT GCGCGGAGGA GTTGTGTTTG TGGACGAAGT ACCGAAAGGT 
1801 CTTACCGGAA AACTCGACGC AAGAAAAATC AGAGAGATCC TCATAAAGGC CAAGAAGGGC 
1861 GGAAAGATCG CCGTGTAATT CTAGAGTCGG GGCGGCCGGC CGCTTCGAGC AGACATGATA 
1921 AGATACATTG ATGAGTTTGG ACAAACCACA ACTAGAATGC AGTGAAAAAA ATGCTTTATT 
1981 TGTGAAATTT GTGATGCTAT TGCTTTATTT GTAACCATTA TAAGCTGCAA TAAACAAGTT 
2041 AACAACAACA ATTGCATTCA TTTTATGTTT CAGGTTCAGG GGGAGGTGTG GGAGGTTTTT 
2101 TAAAGCAAGT AAAACCTCTA CAAATGTGGT AAAATCGATA AGGATCCGTC GACCGATGCC 
2161 CTTGAGAGCC TTCAACCCAG TCAGCTCCTT CCGGTGGGCG CGGGGCATGA CTATCGTCGC 
2221 CGCACTTATG ACTGTCTTCT TTATCATGCA ACTCGTAGGA CAGGTGCCGG CAGCGCTCTT 
2281 CCGCTTCCTC GCTCACTGAC TCGCTGCGCT CGGTCGTTCG GCTGCGGCGA GCGGTATCAG 
2341 CTCACTCAAA GGCGGTAATA CGGTTATCCA CAGAATCAGG GGATAACGCA GGAAAGAACA 
2401 TGTGAGCAAA AGGCCAGCAA AAGGCCAGGA ACCGTAAAAA GGCCGCGTTG CTGGCGTTTT 
2461 TCCATAGGCT CCGCCCCCCT GACGAGCATC ACAAAAATCG ACGCTCAAGT CAGAGGTGGC 
2521 GAAACCCGAC AGGACTATAA AGATACCAGG CGTTTCCCCC TGGAAGCTCC CTCGTGCGCT 
2581 CTCCTGTTCC GACCCTGCCG CTTACCGGAT ACCTGTCCGC CTTTCTCCCT TCGGGAAGCG 
2641 TGGCGCTTTC TCATAGCTCA CGCTGTAGGT ATCTCAGTTC GGTGTAGGTC GTTCGCTCCA 
2701 AGCTGGGCTG TGTGCACGAA CCCCCCGTTC AGCCCGACCG CTGCGCCTTA TCCGGTAACT 
2761 ATCGTCTTGA GTCCAACCCG GTAAGACACG ACTTATCGCC ACTGGCAGCA GCCACTGGTA 
2821 ACAGGATTAG CAGAGCGAGG TATGTAGGCG GTGCTACAGA GTTCTTGAAG TGGTGGCCTA 
2881 ACTACGGCTA CACTAGAAGG ACAGTATTTG GTATCTGCGC TCTGCTGAAG CCAGTTACCT 
2941 TCGGAAAAAG AGTTGGTAGC TCTTGATCCG GCAAACAAAC CACCGCTGGT AGCGGTGGTT 
3001 TTTTTGTTTG CAAGCAGCAG ATTACGCGCA GAAAAAAAGG ATCTCAAGAA GATCCTTTGA 
3061 TCTTTTCTAC GGGGTCTGAC GCTCAGTGGA ACGAAAACTC ACGTTAAGGG ATTTTGGTCA 
3121 TGAGATTATC AAAAAGGATC TTCACCTAGA TCCTTTTAAA TTAAAAATGA AGTTTTAAAT 
3181 CAATCTAAAG TATATATGAG TAAACTTGGT CTGACAGTTA CCAATGCTTA ATCAGTGAGG 
3241 CACCTATCTC AGCGATCTGT CTATTTCGTT CATCCATAGT TGCCTGACTC CCCGTCGTGT 
3301 AGATAACTAC GATACGGGAG GGCTTACCAT CTGGCCCCAG TGCTGCAATG ATACCGCGAG 
3361 ACCCACGCTC ACCGGCTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC 
3421 GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG 
3481 CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA 
3541 TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA 
3601 GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA 
3661 TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA 
3721 ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA 
3781 AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG 
3841 ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG 
3901 GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG 
3961 CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG 
4021 GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC 
4081 TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA 
4141 TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG 
4201 TGCCACCTGA CGCGCCCTGT AGCGGCGCAT TAAGCGCGGC GGGTGTGGTG GTTACGCGCA 
4261 GCGTGACCGC TACACTTGCC AGCGCCCTAG CGCCCGCTCC TTTCGCTTTC TTCCCTTCCT 
4321 TTCTCGCCAC GTTCGCCGGC TTTCCCCGTC AAGCTCTAAA TCGGGGGCTC CCTTTAGGGT 
4381 TCCGATTTAG TGCTTTACGG CACCTCGACC CCAAAAAACT TGATTAGGGT GATGGTTCAC 
4441 GTAGTGGGCC ATCGCCCTGA TAGACGGTTT TTCGCCCTTT GACGTTGGAG TCCACGTTCT 
4501 TTAATAGTGG ACTCTTGTTC CAAACTGGAA CAACACTCAA CCCTATCTCG GTCTATTCTT 
4561 TTGATTTATA AGGGATTTTG CCGATTTCGG CCTATTGGTT AAAAAATGAG CTGATTTAAC 
4621 AAAAATTTAA CGCGAATTTT AACAAAATAT TAACGTTTAC AATTTCCCAT TCGCCATTCA 
4681 GGCTGCGCAA CTGTTGGGAA GGGCGATCGG TGCGGGCCTC TTCGCTATTA CGCCAGCCCA 
4741 AGCTACCATG ATAAGTAAGT AATATTAAGG TACGGGAGGT ACTTGGAGCG GCCGCAATAA 
4801 AATATCTTTA TTTTCATTAC ATCTGTGTGT TGGTTTTTTG TGTGAATCGA TAGTACTAAC 
4861 ATACGCTCTC CATCAAAACA AAACGAAACA AAACAAACTA GCAAAATAGG CTGTCCCCAG 
4921 TGCAAGTGCA GGTGCCAGAA CATTTCTCTA TCGATA