LOCUS pLEW82v4 6641 bp DNA circular 23-JUL-2008 FEATURES Location/Qualifiers misc_feature complement(1..713) /function="rRNA Spacer" protein_bind 741..813 /function="Triple Tet Operator" promoter 720..739 /function="T7 Promoter" /note="T7 promoter & initiation region. Promoter-Op-Luc cassette orig from Lew19" 5'UTR 822..877 /function="GPEET upstream" /note="to splice site. Note that this is truncated at the upstream SmaI site compared to the GPEET upstream region in pLew20" 5'UTR 878..908 /function="GPEET 5´ UTR" CDS 983..2632 /function="LUCIFERASE" promoter 2811..2830 /function="10% T7 promoter" 3'UTR 2835..2987 /function="ALD 'short' 3´ UTR" /note="Perfect match with 3480 to 3612 in X52586" /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) " 5'UTR 2988..3102 /function="ACT 5´ UTR" /note="actin-derived sas; used in all HD330, 430 & Lew5 descendants" CDS 3112..3483 /function="BLE" 3'UTR 3538..3825 /function="ACT 3´ UTR" /note="actin 3'UTR- same as what's used in all HD330, 430 and Lew5 descendants" terminator 3832..3964 /function="T7 Term" /note="132bp Pet3a fragment (originally PCR'd), then subcloned from published HD216, bearing T7 terminator" terminator 3965..4097 /function="T7 Term" /note="132 bp Pet3a fragment (originally PCR'd then subcloned from published HD216) bearing T7 terminator" CDS 4786..5643 /function="LACTAMASE" CDS 940..981 /note="Extraneous CDS" conflict 589..594 /note="The Sac II site indicated in the sequence is missing." BASE COUNT 1711 a 1569 c 1697 g 1664 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 TAGGGAAGCG ACGCTTGCCC 481 TAGTCCCACG AGTAACCAAG ACTCCAAAAG CCTTTCTGGC ACAGAGAGCG AGCCGAAATG 541 GAAAAGAGAA ACAATGCCTG CACTAACACT ACTGAGCGAT TCGCCTCGCC GCGGAGGACC 601 GAATACTAAT AACGACACTT GCGGTCAAAA AGTAGAAGAA CAAATGCTCA ACGATGAGTG 661 AATCAGGTTA GGGTAGTTGG AAAATTATAC AGAATGTCTT TGGCAACACA CCGGCTGATT 721 AATACGACTC ACTATAGGGA GATCTCCCTA TCAGTGATAG AGACATCCGC TCCCTATCAG 781 TGATAGAGAC GGTAGCTCCC TATCAGTGAT AGAGATCTCC CGGGCTGCAC GCGCCTTCGA 841 GTTTTTTTTC CTTTTCCCCA TTTTTTTCAA CTTGAAGACT TCAATTACAC CAAAAAGTAA 901 AATTCACAAG CTTGGAATTC CTTTGTGTTA CATTCTTGAA TGTCGCTCGC AGTGACATTA 961 GCATTCCGGT ACTGTTGGTA AAATGGAAGA CGCCAAAAAC ATAAAGAAAG GCCCGGCGCC 1021 ATTCTATCCT CTAGAGGATG GAACCGCTGG AGAGCAACTG CATAAGGCTA TGAAGAGATA 1081 CGCCCTGGTT CCTGGAACAA TTGCTTTTAC AGATGCACAT ATCGAGGTGA ACATCACGTA 1141 CGCGGAATAC TTCGAAATGT CCGTTCGGTT GGCAGAAGCT ATGAAACGAT ATGGGCTGAA 1201 TACAAATCAC AGAATCGTCG TATGCAGTGA AAACTCTCTT CAATTCTTTA TGCCGGTGTT 1261 GGGCGCGTTA TTTATCGGAG TTGCAGTTGC GCCCGCGAAC GACATTTATA ATGAACGTGA 1321 ATTGCTCAAC AGTATGAACA TTTCGCAGCC TACCGTAGTG TTTGTTTCCA AAAAGGGGTT 1381 GCAAAAAATT TTGAACGTGC AAAAAAAATT ACCAATAATC CAGAAAATTA TTATCATGGA 1441 TTCTAAAACG GATTACCAGG GATTTCAGTC GATGTACACG TTCGTCACAT CTCATCTACC 1501 TCCCGGTTTT AATGAATACG ATTTTGTACC AGAGTCCTTT GATCGTGACA AAACAATTGC 1561 ACTGATAATG AATTCCTCTG GATCTACTGG GTTACCTAAG GGTGTGGCCC TTCCGCATAG 1621 AACTGCCTGC GTCAGATTCT CGCATGCCAG AGATCCTATT TTTGGCAATC AAATCATTCC 1681 GGATACTGCG ATTTTAAGTG TTGTTCCATT CCATCACGGT TTTGGAATGT TTACTACACT 1741 CGGATATTTG ATATGTGGAT TTCGAGTCGT CTTAATGTAT AGATTTGAAG AAGAGCTGTT 1801 TTTACGATCC CTTCAGGATT ACAAAATTCA AAGTGCGTTG CTAGTACCAA CCCTATTTTC 1861 ATTCTTCGCC AAAAGCACTC TGATTGACAA ATACGATTTA TCTAATTTAC ACGAAATTGC 1921 TTCTGGGGGC GCACCTCTTT CGAAAGAAGT CGGGGAAGCG GTTGCAAAAC GCTTCCATCT 1981 TCCAGGGATA CGACAAGGAT ATGGGCTCAC TGAGACTACA TCAGCTATTC TGATTACACC 2041 CGAGGGGGAT GATAAACCGG GCGCGGTCGG TAAAGTTGTT CCATTTTTTG AAGCGAAGGT 2101 TGTGGATCTG GATACCGGGA AAACGCTGGG CGTTAATCAG AGAGGCGAAT TATGTGTCAG 2161 AGGACCTATG ATTATGTCCG GTTATGTAAA CAATCCGGAA GCGACCAACG CCTTGATTGA 2221 CAAGGATGGA TGGCTACATT CTGGAGACAT AGCTTACTGG GACGAAGACG AACACTTCTT 2281 CATAGTTGAC CGCTTGAAGT CTTTAATTAA ATACAAAGGA TATCAGGTGG CCCCCGCTGA 2341 ATTGGAATCG ATATTGTTAC AACACCCCAA CATCTTCGAC GCGGGCGTGG CAGGTCTTCC 2401 CGACGATGAC GCCGGTGAAC TTCCCGCCGC CGTTGTTGTT TTGGAGCACG GAAAGACGAT 2461 GACGGAAAAA GAGATCGTGG ATTACGTCGC CAGTCAAGTA ACAACCGCGA AAAAGTTGCG 2521 CGGAGGAGTT GTGTTTGTGG ACGAAGTACC GAAAGGTCTT ACCGGAAAAC TCGACGCAAG 2581 AAAAATCAGA GAGATCCTCA TAAAGGCCAA GAAGGGCGGA AAGTCCAAAT TGTAAAATGT 2641 AACTGTATTC AGCGATGACG AAATTCTTAG CTATTGTAAT ATTATATGCA AATTGATGAA 2701 TGGTAATTTT GTAATTGTGG GTCACTGTAC TATTTTAACG AATAATAAAA TCAGGTATAG 2761 GTAACTAAAA AGGAATTCGA GCTCGGATCC AAGCTTCTAG ACTCGAGCAT TAATACGTCT 2821 CACTATAGGG AGAGGATCCT GCCCATTTAG TTGGCTTTTC CCTTGTCTCG TGTCTTTTCC 2881 GTGGAAAGGT TCCCGGAGTA ATCTGATGGC ACAGCAGGGA GGTGCGCCTG CAGGTTGGTT 2941 AGGAAGGGGG GATGATGTAA AAGAAGAAAA TGGGGGGATT CGAGCCCGGG CACAGCAAGG 3001 TCTTCTGAAA TTCATGTTTT TTTTTTTTTT ACTCTGCATT GCAGTCTCCG CTCTTATTTA 3061 GTTTTGCTTT ACGTAAGGTC TCGTTGCTGC CATAAAATAA GCTATATCAC CATGGCCAAG 3121 TTGACCAGTG CCGTTCCGGT GCTCACCGCG CGCGACGTCG CCGGAGCGGT CGAGTTCTGG 3181 ACCGACCGGC TCGGGTTCTC CCGGGACTTC GTGGAGGACG ACTTCGCCGG TGTGGTCCGG 3241 GACGACGTGA CCCTGTTCAT CAGCGCGGTC CAGGACCAGG TGGTGCCGGA CAACACCCTG 3301 GCCTGGGTGT GGGTGCGCGG CCTGGACGAG CTGTACGCCG AGTGGTCGGA GGTCGTGTCC 3361 ACGAACTTCC GGGACGCCTC CGGGCCGGCC ATGACCGAGA TCGGCGAGCA GCCGTGGGGG 3421 CGGGAGTTCG CCCTGCGCGA CCCGGCCGGC AACTGCGTGC ACTTCGTGGC CGAGGAGCAG 3481 GACTGACCGA CGCCGACCAA CACCGCCGGT CCGACGGCGG CCCACGGGTC CCAGGCCTCG 3541 GAGATCCTAA CACCGGGTTG TGTGGCCAAA ATTGTTCTGT AGTCGCTGTG AGTTGACACG 3601 GCTAGTGCTT ATGATTTTCC TCGCGTGTGG TGCCTGTACT CAGCCCTATG CCTTATTTGC 3661 AACACATTTA CGTACAGCGC ACAAGAGAAG AGAAGATCAC TTGAAGATAA TAAATATAGG 3721 GTTGTAGGCA TCTTGTTTAA CTCAAATTTT CTCGTCTTGG TGTGTCGACA TGATTGAAAT 3781 AGTGCCACCA GTTGTGTTTG ATGCGTTTGT TATCTATGCA GTATTCTGCA GGCTGCTAAC 3841 AAAGCCCGAA AGGAAGCTGA GTTGGCTGCT GCCACCGCTG AGCAATAACT AGCATAACCC 3901 CTTGGGGCCT CTAAACGGGT CTTGAGGGGT TTTTTGCTGA AAGGAGGAAC TATATCCGGA 3961 TAGGGCTGCT AACAAAGCCC GAAAGGAAGC TGAGTTGGCT GCTGCCACCG CTGAGCAATA 4021 ACTAGCATAA CCCCTTGGGG CCTCTAAACG GGTCTTGAGG GGTTTTTTGC TGAAAGGAGG 4081 AACTATATCC GGATAGGGCC CCTGCAGGCA TGCAAGCTAG CCAGATCCTC TACGCCGGAC 4141 GCATCGTGGC CGGCATCACC GGCGCCACAG GTGCGGTTGC TGGCGCCTAT ATCGCCGACA 4201 TCACCGATGG GGAAGATCGG GCTCGCCACT TCGGGCTCAT GAGCGCTTGT TTCGGCGTGG 4261 GTATGGTGGC AGGCCCCGTG GCCGGGGGAC TGTTGGGCGC CATCTCCTTG CACCATTCCT 4321 TGCGGCGGCG GTGCTCAACG GCCTCAACCT ACTACTGGGC TGCTTCCTAA TGCAGGAGTC 4381 GCATAAGGGA GAGCGTCGAT ATGGTGCACT CTCAGTACAA TCTGCTCTGA TGCCGCATAG 4441 TTAAGCCAGC CCCGACACCC GCCAACACCC GCTGACGCGC CCTGACGGGC TTGTCTGCTC 4501 CCGGCATCCG CTTACAGACA AGCTGTGACC GTCTCCGGGA GCTGCATGTG TCAGAGGTTT 4561 TCACCGTCAT CACCGAAACG CGCGAGACGA AAGGGCCTCG TGATACGCCT ATTTTTATAG 4621 GTTAATGTCA TGATAATAAT GGTTTCTTAG ACGTCAGGTG GCACTTTTCG GGGAAATGTG 4681 CGCGGAACCC CTATTTGTTT ATTTTTCTAA ATACATTCAA ATATGTATCC GCTCATGAGA 4741 CAATAACCCT GATAAATGCT TCAATAATAT TGAAAAAGGA AGAGTATGAG TATTCAACAT 4801 TTCCGTGTCG CCCTTATTCC CTTTTTTGCG GCATTTTGCC TTCCTGTTTT TGCTCACCCA 4861 GAAACGCTGG TGAAAGTAAA AGATGCTGAA GATCAGTTGG GTGCACGAGT GGGTTACATC 4921 GAACTGGATC TCAACAGCGG TAAGATCCTT GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA 4981 ATGATGAGCA CTTTTAAAGT TCTGCTATGT GGCGCGGTAT TATCCCGTAT TGACGCCGGG 5041 CAAGAGCAAC TCGGTCGCCG CATACACTAT TCTCAGAATG ACTTGGTTGA GTACTCACCA 5101 GTCACAGAAA AGCATCTTAC GGATGGCATG ACAGTAAGAG AATTATGCAG TGCTGCCATA 5161 ACCATGAGTG ATAACACTGC GGCCAACTTA CTTCTGACAA CGATCGGAGG ACCGAAGGAG 5221 CTAACCGCTT TTTTGCACAA CATGGGGGAT CATGTAACTC GCCTTGATCG TTGGGAACCG 5281 GAGCTGAATG AAGCCATACC AAACGACGAG CGTGACACCA CGATGCCTGT AGCAATGGCA 5341 ACAACGTTGC GCAAACTATT AACTGGCGAA CTACTTACTC TAGCTTCCCG GCAACAATTA 5401 ATAGACTGGA TGGAGGCGGA TAAAGTTGCA GGACCACTTC TGCGCTCGGC CCTTCCGGCT 5461 GGCTGGTTTA TTGCTGATAA ATCTGGAGCC GGTGAGCGTG GGTCTCGCGG TATCATTGCA 5521 GCACTGGGGC CAGATGGTAA GCCCTCCCGT ATCGTAGTTA TCTACACGAC GGGGAGTCAG 5581 GCAACTATGG ATGAACGAAA TAGACAGATC GCTGAGATAG GTGCCTCACT GATTAAGCAT 5641 TGGTAACTGT CAGACCAAGT TTACTCATAT ATACTTTAGA TTGATTTAAA ACTTCATTTT 5701 TAATTTAAAA GGATCTAGGT GAAGATCCTT TTTGATAATC TCATGACCAA AATCCCTTAA 5761 CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA 5821 GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG 5881 GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC 5941 AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT AGTTAGGCCA CCACTTCAAG 6001 AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC TGTTACCAGT GGCTGCTGCC 6061 AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG 6121 CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC 6181 ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC CGAAGGGAGA 6241 AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG GAGAGCGCAC GAGGGAGCTT 6301 CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT TTCGCCACCT CTGACTTGAG 6361 CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG 6421 GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATGTTCTT TCCTGCGTTA 6481 TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC CGCTCGCCGC 6541 AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG CGGAAGAGCG CCCAATACGC 6601 AAACCGCCTC TCCCCGCGCG TTGGCCGATT CATTAATGCA G //