LOCUS Plew111-BSD-(pFlan1) 4662 bp DNA circular 25-JUL-2008 FEATURES Location/Qualifiers misc_feature complement(join(4655..4662,1..712)) /function="rRNA Spacer" promoter 719..738 /function="T7 Promoter" /note="T7 promoter & initiation region. Bgl II starts @ +4. 1 TetR Operator in Bgl II. Promoter-Op-Luc cassette orig from Lew19" protein_bind 744..762 /function="Tet Operator" misc_feature 769..824 /function="GPEET SAS" /note="to splice site" 5'UTR 825..855 /function="GPEET 5´ UTR" 3'UTR 899..1051 /function="ALD 3´ UTR" 5'UTR 1057..1251 /function="ALD 5' UTR & SAS" /note="5926 to 6120 in X52586" CDS 1255..1653 /function="BSD" /note="Blasticidin S deaminase. Not sure where Fabian got the BSD-UTR cassette from" 3'UTR 1703..1835 /function="ALD 'short' 3´ UTR" /note="truncated aldolase 3'UTR originally derived from pHD330 (about 6x lower BF expression than the full-length ALD UTR in pLew100, but no difference in expression levels in PF, between long and short UTRs) " /note="perfect match with 3480 to 3612 in X52586" terminator 1847..1987 /function="T7 Term" /note="Pet3a fragment (originally PCR'd), then subcloned from published HD216, bearing T7 terminator" terminator 1988..2128 /function="T7 Term" /note="Pet3a fragment (originally PCR'd then subcloned from published HD216) bearing T7 terminator" CDS 2807..3664 /function="LACTAMASE" BASE COUNT 1176 a 1156 c 1210 g 1120 t ORIGIN 1 GAATTCGAGC TCATATAGTT GGTATGTATT CTAATTCCAG ACTACTGGCG TGGATAAACA 61 TGTCCCCTGA TTAAAAGGAA AGATTCCATA GCCCATTAGT GCAAAGATAA TTGGTACACC 121 GTCAAAAACA CATGGGAGAC CACACGATAC GACACTGTGA CCGTAGCATC AATTCCGCAC 181 TCGATATAAC TCTATCGAAG AACTTCCATG GTACAAACTG GTCGCCACGG TCTGCTCCAC 241 ACGGAGATCA TCGTATCATT TTTATCGATA GCGGCCGCTA TCGATGTATG CCTTGGCCCT 301 GATGGCATGC CAATTTCACT ACAACGGACT ATGTGGACCC CGTTATCATG GAAATGCGCT 361 AGTTGGAGGA AGTTAGACCG CGCCGGAAAA GAGAGGGGTA GAGAAAATGA CAACTTGGAA 421 GATATCCACA CACGCACGGT GAAACGTTAG CAACAATTAT TAGGGAAGCA CGCTTGCCCT 481 AGTCCCACGA GTAACCAAGA CTCCAAAAGC CTTTCTGGCA CAGAGAGCGA GCCGAAATGG 541 AAAAGAGAAA CAATGCCTGC ACTAACACTA CTGAGCGATT CGCCTCGCCG CGGAGGACCG 601 AATACTAATA ACGACACTTG CGGTCAAAAA GTAGAAGAAC AAATGCTCAA CGATGAGTGA 661 ATCAGGTTAG GGTAGTTGGA AAATTATACA GAATGTCTTT GGCAACACAC CGGCTGATTA 721 ATACGACTCA CTATAGGGAG ATCTCCCTAT CAGTGATAGA GATCTCCCGG GCTGCACGCG 781 CCTTCGAGTT TTTTTTCCTT TTCCCCATTT TTTTCAACTT GAAGACTTCA ATTACACCAA 841 AAAGTAAAAT TCACAAGCTT CGTCGCGACC AGTGTGCTGG CTGCTGGCCA GCACACTGGA 901 TCCTGCCCAT TTAGTTGGCT TTTCCCTTGT CTCGTGTCTT TTCCGTGGAA AGGTTCCCGG 961 AGTAATCTGA TGGCACAGCA GGGAGGTGCG CCTGCAGGTT GGTTAGGAAG GGGGGATGAT 1021 GTAAAAGAAG AAAATGGGGG GATTCGAGCC CcTCGAGGGT GCTCAAGCTG TGTAGCGCAC 1081 GCGTTTCCTT ACATATTTCT CTAACAGGCA CGGAAGCCTA ACAAATACAC TTGGCTTATT 1141 TTTTTGCCCC CTCATGTCTT GTACAAATAT TTGCGATAGC TTAGCTATCA GCCACATTAA 1201 TCAAACAAGT ATACCAACAA GCCCGAAAAC AGAAACTCAA CTGCAACGAA GCTCATGGCC 1261 AAGCCTTTGT CTCAAGAAGA ATCCACCCTC ATTGAAAGAG CAACGGCTAC AATCAACAGC 1321 ATCCCCATCT CTGAAGACTA CAGCGTCGCC AGCGCAGCTC TCTCTAGCGA CGGCCGCATC 1381 TTCACTGGTG TCAATGTATA TCATTTTACT GGGGGACCTT GTGCAGAACT CGTGGTGCTG 1441 GGCACTGCTG CTGCTGCGGC AGCTGGCAAC CTGACTTGTA TCGTCGCGAT CGGAAATGAG 1501 AACAGGGGCA TCTTGAGCCC CTGCGGACGG TGCCGACAGG TGCTTCTCGA TCTGCATCCT 1561 GGGATCAAAG CCATAGTGAA GGACAGTGAT GGACAGCCGA CGGCAGTTGG GATTCGTGAA 1621 TTGCTGCCCT CTGGTTATGT GTGGGAGGGC TAAGCACTTC GTGGCCGAGG AGCAGGACTG 1681 AGAATTGATC CTGCCCATTT AGTTAGTTGG CTTTTCCCTT GTCTCGTGTC TTTTCCGTGG 1741 AAAGGTTCCC GGAGTAATCT GATGGCACAG CAGGGAGGTG CGCCTGCAGG TTGGTTAGGA 1801 AGGGGGGATG ATGTAAAAGA AGAAAATGGG GGGATTCGAG CCCcctCTGC AGGCTGCTAA 1861 CAAAGCCCGA AAGGAAGCTG AGTTGGCTGC TGCCACCGCT GAGCAATAAC TAGCATAACC 1921 CCTTGGGGCC TCTAAACGGG TCTTGAGGGG TTTTTTGCTG AAAGGAGGAA CTATATCCGG 1981 ATAGGGCTGC TAACAAAGCC CGAAAGGAAG CTGAGTTGGC TGCTGCCACC GCTGAGCAAT 2041 AACTAGCATA ACCCCTTGGG GCCTCTAAAC GGGTCTTGAG GGGTTTTTTG CTGAAAGGAG 2101 GAACTATATC CGGATAGGGC CCCTGCAGGC ATGCAAGCTA GCCAGATCCT CTACGCCGGA 2161 CGCATCGTGG CCGGCATCAC CGGCGCCACA GGTGCGGTTG CTGGCGCCTA TATCGCCGAC 2221 ATCACCGATG GGGAAGATCG GGCTCGCCAC TTCGGGCTCA TGAGCGCTTG TTTCGGCGTG 2281 GGTATGGTGG CAGGCCCCGT GGCCGGGGGA CTGTTGGGCG CCATCTCCTT GCACCATTCC 2341 TTGCGGCGGC GGTGCTCAAC GGCCTCAACC TACTACTGGG CTGCTTCCTA ATGCAGGAGT 2401 CGCATAAGGG AGAGCGTCGA TATGGTGCAC TCTCAGTACA ATCTGCTCTG ATGCCGCATA 2461 GTTAAGCCAG CCCCGACACC CGCCAACACC CGCTGACGCG CCCTGACGGG CTTGTCTGCT 2521 CCCGGCATCC GCTTACAGAC AAGCTGTGAC CGTCTCCGGG AGCTGCATGT GTCAGAGGTT 2581 TTCACCGTCA TCACCGAAAC GCGCGAGACG AAAGGGCCTC GTGATACGCC TATTTTTATA 2641 GGTTAATGTC ATGATAATAA TGGTTTCTTA GACGTCAGGT GGCACTTTTC GGGGAAATGT 2701 GCGCGGAACC CCTATTTGTT TATTTTTCTA AATACATTCA AATATGTATC CGCTCATGAG 2761 ACAATAACCC TGATAAATGC TTCAATAATA TTGAAAAAGG AAGAGTATGA GTATTCAACA 2821 TTTCCGTGTC GCCCTTATTC CCTTTTTTGC GGCATTTTGC CTTCCTGTTT TTGCTCACCC 2881 AGAAACGCTG GTGAAAGTAA AAGATGCTGA AGATCAGTTG GGTGCACGAG TGGGTTACAT 2941 CGAACTGGAT CTCAACAGCG GTAAGATCCT TGAGAGTTTT CGCCCCGAAG AACGTTTTCC 3001 AATGATGAGC ACTTTTAAAG TTCTGCTATG TGGCGCGGTA TTATCCCGTA TTGACGCCGG 3061 GCAAGAGCAA CTCGGTCGCC GCATACACTA TTCTCAGAAT GACTTGGTTG AGTACTCACC 3121 AGTCACAGAA AAGCATCTTA CGGATGGCAT GACAGTAAGA GAATTATGCA GTGCTGCCAT 3181 AACCATGAGT GATAACACTG CGGCCAACTT ACTTCTGACA ACGATCGGAG GACCGAAGGA 3241 GCTAACCGCT TTTTTGCACA ACATGGGGGA TCATGTAACT CGCCTTGATC GTTGGGAACC 3301 GGAGCTGAAT GAAGCCATAC CAAACGACGA GCGTGACACC ACGATGCCTG TAGCAATGGC 3361 AACAACGTTG CGCAAACTAT TAACTGGCGA ACTACTTACT CTAGCTTCCC GGCAACAATT 3421 AATAGACTGG ATGGAGGCGG ATAAAGTTGC AGGACCACTT CTGCGCTCGG CCCTTCCGGC 3481 TGGCTGGTTT ATTGCTGATA AATCTGGAGC CGGTGAGCGT GGGTCTCGCG GTATCATTGC 3541 AGCACTGGGG CCAGATGGTA AGCCCTCCCG TATCGTAGTT ATCTACACGA CGGGGAGTCA 3601 GGCAACTATG GATGAACGAA ATAGACAGAT CGCTGAGATA GGTGCCTCAC TGATTAAGCA 3661 TTGGTAACTG TCAGACCAAG TTTACTCATA TATACTTTAG ATTGATTTAA AACTTCATTT 3721 TTAATTTAAA AGGATCTAGG TGAAGATCCT TTTTGATAAT CTCATGACCA AAATCCCTTA 3781 ACGTGAGTTT TCGTTCCACT GAGCGTCAGA CCCCGTAGAA AAGATCAAAG GATCTTCTTG 3841 AGATCCTTTT TTTCTGCGCG TAATCTGCTG CTTGCAAACA AAAAAACCAC CGCTACCAGC 3901 GGTGGTTTGT TTGCCGGATC AAGAGCTACC AACTCTTTTT CCGAAGGTAA CTGGCTTCAG 3961 CAGAGCGCAG ATACCAAATA CTGTCCTTCT AGTGTAGCCG TAGTTAGGCC ACCACTTCAA 4021 GAACTCTGTA GCACCGCCTA CATACCTCGC TCTGCTAATC CTGTTACCAG TGGCTGCTGC 4081 CAGTGGCGAT AAGTCGTGTC TTACCGGGTT GGACTCAAGA CGATAGTTAC CGGATAAGGC 4141 GCAGCGGTCG GGCTGAACGG GGGGTTCGTG CACACAGCCC AGCTTGGAGC GAACGACCTA 4201 CACCGAACTG AGATACCTAC AGCGTGAGCT ATGAGAAAGC GCCACGCTTC CCGAAGGGAG 4261 AAAGGCGGAC AGGTATCCGG TAAGCGGCAG GGTCGGAACA GGAGAGCGCA CGAGGGAGCT 4321 TCCAGGGGGA AACGCCTGGT ATCTTTATAG TCCTGTCGGG TTTCGCCACC TCTGACTTGA 4381 GCGTCGATTT TTGTGATGCT CGTCAGGGGG GCGGAGCCTA TGGAAAAACG CCAGCAACGC 4441 GGCCTTTTTA CGGTTCCTGG CCTTTTGCTG GCCTTTTGCT CACATGTTCT TTCCTGCGTT 4501 ATCCCCTGAT TCTGTGGATA ACCGTATTAC CGCCTTTGAG TGAGCTGATA CCGCTCGCCG 4561 CAGCCGAACG ACCGAGCGCA GCGAGTCAGT GAGCGAGGAA GCGGAAGAGC GCCCAATACG 4621 CAAACCGCCT CTCCCCGCGC GTTGGCCGAT TCATTAATGC AG //