pBUF has a 2-kb Ubiquitin promoter, which drives a moderate level of expression in all tissues; heat-shock causes about 3X increased transcription (Lee et al., MCB 8:4727, 1988). The ATG corresponding to the initial codon is followed by sequences encoding the FLAG epitope. Coding sequences can be cloned in-frame using the sites in the polylinker. Alternatively, one can use the NcoI site that overlaps the ATG, to produce a non-tagged polypeptide. Note: The anti-FLAG Ab's (M2 and M5) recognize endogenous Drosophila proteins with Mr ca. 50 kD.
pBUF Features |
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1145-1276 |
exon 1 (non-coding) |
2032 |
start of exon 2 |
2043-2045 |
initial ATG |
2043-2069 |
FLAG tag |
pBUF Sequence
GGTACCGGGCCCCCCTCGAGGTCGACTCTAGAGGATCTTGTCGCCGGAACGCAGCGACAGAGAT TCCAATGTGTCCGTATCTTTCAGGCTTTTGCCCTTCAGTTCCAGACGAAGCGACTGGCGATTCG CGTGTGGGGTCTGCTTCAGGGTCTTGTGAATTAGGGCGCGCAGATCGCCGATGGGCGTGGCGCC GGAGGGCACCTTCACCTTGCCGTACGGCTTGCTGTTCTTCGCGTTCAAAATCTCCAGCTCCATT TTGCTTTCGGTGCGCTTGCAATCAGTACTGTCCAAAATCGAAAATCGCCGAACCGTAGTGTGAC CGTGCGGGGCTCTGCGAAAATAAACTTTTTTAGGTATATGGCCACACACGGGGAAAGCACAGTG GATTATATGTTTTAATATTATAATATGCAGGTTTTCATTACTTATCCAGATGTAAGCCCACTTA AAGCGATTTAACAATTATTTGCCGAAAGAGTATAAACAAATTTCACTTAAAAATGGATTAAGAA AAGCTTGTGTAAGATTATGCGCAGCGTTGCCAGATAGCTCCATTTAAAACACTTCAAAAACAAT AAGTTTTGAAAATATATACATAAATAGCAGTCGTTGCCGCAACGCTCAACACATCACACTTTTA AAACACCCTTTACCTACACAGAATTACTTTTTAAATTTCCAGTCAAGCTGCGAGTTTCAAAATT ATAGCCGGTAGAGAAGACAGTGCTATTTCAAAAGCAAACTAACAAGGGTCTTAAATTCCAAAAC ACCAATCCTAACAAGCCTTGGACTTTTGTAAGTTTAGATCAAAGGTGGCATTGCATTCAATGTC ATGGTAAGAAGTAGGTCGTCTAGGTAGAAATCCTCATTCAGCCGGTCAAGTCAGTACGAGAAAG GTCTCAATTTGAAATTGTCTTAAAAATATTTTATTGTTTTGTACTGTGGTGAGTTTAAACGAAA AACACAAAAAAAAAGTGATACACAGAAATCATAAAAAATTTTAATACAAGGTATTCGTACGTAT CAAAAACATTTCGGCACAATTTTTTTTCTCTGTACTAAAGTGTTACGAACACTACGGTATTTTT TAGTGATTTTCAACGGACACCGAAGGTATATAAACAGCGTTCGCGAACGGTCGCCTTCAAAACC AATTGACATTTGCAGCAGCAAGTACAAGCAGAAAGTAAAGCGCAATCAGCGAAAAATTTATACT TAATTGTTGGTGATTAAAGTACAATTAAAAGAACATTCTCGAAAGTCACAAGAAACGTAAGTTT TTAACTCGCTGTTACCAATTAGTAATAAGAGCAACAAGACGTTGAGTAATTTCAAGAAAAACTG CATTTCAAGGTCTTTGTTCGGCCATTTTTTTTTTATTCAACGCTCTACGTAATTACAAAATAAG AAATTGGCAGCCACGCATCTTGTTTTCCCAATCAATTGGCATCAAAACGCAAACAAATCTATAA ATAAAACTTGCGTGTTGATTTTCGCCAAGATTTATTGGCAAATTGTGAAATTCGCAGTGACGCA TTTGAAAATTCGAGAAATCACGAACGCACTCGAGCATTTGTGTGCATGTTATTAGTTAGTTAGT TCTTTGCTTAATTGAAGTATTTTACCAACGAAATCCACTTATTTTTAGCTGAAATAGAGTAGGT TGCTTGAAACGAAAGCCACGTCTGGAAAATTTCTTATTGCTTAGTAGTTGTGACGTCACCATAT ACACACAAAATAATGTGTATGCATGCGTTTCAGCTGTGTATATATACATGCACACACTCGCATT ATGAAAACGATGACGAGCAACGGAACAGGTTTCTCAACTACCTTTGTTCCTGTTTCTTCGCTTT CCTTTGTTCCAATATTCGTAGAGGGTTAATAGGGGTTTCTCAACAAAGTTGGCGTCGATAAATA AGTTTCCCATTTTTATTCCCCAGCCAGGAAGTTAGTTTCAATAGTTTTGTAATTTCAACGAAAC TCATTTGATTTCGTACTAATTTTCCACATCTCTATTTTGACCCGCAGAATAATCCACCATGGAC TACAAGGACGACGATGACAAGGCCTCATCGATAAGCTTGATATCGAATTCCTGCAGCCCGGGCA GGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCAATTCGCCCTATAGTGAGTC GTATTACAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAA CTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCG ATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGCGACGCGCCCTGTAGCGGCGC ATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCG CCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTC TAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACT TGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACG TTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCT CGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCT GATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCCCAGGTGGCA CTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTA TCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGT ATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTC ACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACAT CGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATG ATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGC AACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAAC ACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACA ACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAA CGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGC GAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAG GACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGA GCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTT ATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTG CCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTT AAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAA ATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTT CTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGC GGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGA GCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTG TAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAA GTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGA ACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTAC AGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTAT AGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGC GGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTT TGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAG TGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGG AAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCA CGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACT CATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCG GATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTCGGAATTAACCCTCA CTAAAGGGAACAAAAGCTG
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