LOCUS pCO57 5423 bp DNA circular 17-JUL-2008 FEATURES Location/Qualifiers misc_feature complement(join(5416..5423,1..712)) /function="rRNA Spacer" /note="" promoter 719..738 /function="T7 Promoter" /note="T7 promoter & initiation region. Promoter-Op-Luc cassette orig from Lew19" protein_bind 744..762 /note="Tet Operator" /note="1TetR Operator" promoter 870..881 /note="Linker peptide" CDS 882..1595 /function="eGFP CDS" 5'UTR 769..824 /note="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 825..855 /function="GPEET 5´ UTR" CDS 1596..1607 /function="Linker peptide" 3'UTR 1617..1769 /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) " CDS 1770..1884 /note="ACT upstream" /note="actin-derived sas; used in all HD330, 430 & Lew5 descendants" CDS 1894..2265 /function="BLE" 3'UTR 2320..2607 /function="ACT 3´ UTR" /note="actin 3'UTR- same as what's used in all HD330, 430 and Lew5 descendants" terminator 2614..2746 /function="T7 Term" /note="132bp Pet3a fragment (originally PCR'd), then subcloned from published HD216, bearing T7 terminator" terminator 2747..2879 /function="T7 Term" /note="132 bp Pet3a fragment (originally PCR'd then subcloned from published HD216) bearing T7 terminator" CDS 3568..4425 /function="Beta Lactamase" BASE COUNT 1315 a 1419 c 1446 g 1243 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 gacgttcata tgcagctgAC CATGGTGAGC AAGGGCGAGG 901 AGCTGTTCAC CGGGGTGGTG CCCATCCTGG TCGAGCTGGA CGGCGACGTA AACGGCCACA 961 AGTTCAGCGT GTCCGGCGAG GGCGAGGGCG ATGCCACCTA CGGCAAGCTG ACCCTGAAGT 1021 TCATCTGCAC CACCGGCAAG CTGCCCGTGC CCTGGCCCAC CCTCGTGACC ACCCTGACCT 1081 ACGGCGTGCA GTGCTTCAGC CGCTACCCCG ACCACATGAA GCAGCACGAC TTCTTCAAGT 1141 CCGCCATGCC CGAAGGCTAC GTCCAGGAGC GCACCATCTT CTTCAAGGAC GACGGCAACT 1201 ACAAGACCCG CGCCGAGGTG AAGTTCGAGG GCGACACCCT GGTGAACCGC ATCGAGCTGA 1261 AGGGCATCGA CTTCAAGGAG GACGGCAACA TCCTGGGGCA CAAGCTGGAG TACAACTACA 1321 ACAGCCACAA CGTCTATATC ATGGCCGACA AGCAGAAGAA CGGCATCAAG GTGAACTTCA 1381 AGATCCGCCA CAACATCGAG GACGGCAGCG TGCAGCTCGC CGACCACTAC CAGCAGAACA 1441 CCCCCATCGG CGACGGCCCC GTGCTGCTGC CCGACAACCA CTACCTGAGC ACCCAGTCCG 1501 CCCTGAGCAA AGACCCCAAC GAGAAGCGCG ATCACATGGT CCTGCTGGAG TTCGTGACCG 1561 CCGCCGGGAT CACTCTCGGC ATGGACGAGC TGTACcccgg gtgttaacat taatagGATC 1621 CTGCCCATTT AGTTGGCTTT TCCCTTGTCT CGTGTCTTTT CCGTGGAAAG GTTCCCGGAG 1681 TAATCTGATG GCACAGCAGG GAGGTGCGCC TGCAGGTTGG TTAGGAAGGG GGGATGATGT 1741 AAAAGAAGAA AATGGGGGGA TTCGAGCCCG GGCACAGCAA GGTCTTCTGA AATTCATGTT 1801 TTTTTTTTTT TTACTCTGCA TTGCAGTCTC CGCTCTTATT TAGTTTTGCT TTACGTAAGG 1861 TCTCGTTGCT GCCATAAAAT AAGCTATATC ACCATGGCCA AGTTGACCAG TGCCGTTCCG 1921 GTGCTCACCG CGCGCGACGT CGCCGGAGCG GTCGAGTTCT GGACCGACCG GCTCGGGTTC 1981 TCCCGGGACT TCGTGGAGGA CGACTTCGCC GGTGTGGTCC GGGACGACGT GACCCTGTTC 2041 ATCAGCGCGG TCCAGGACCA GGTGGTGCCG GACAACACCC TGGCCTGGGT GTGGGTGCGC 2101 GGCCTGGACG AGCTGTACGC CGAGTGGTCG GAGGTCGTGT CCACGAACTT CCGGGACGCC 2161 TCCGGGCCGG CCATGACCGA GATCGGCGAG CAGCCGTGGG GGCGGGAGTT CGCCCTGCGC 2221 GACCCGGCCG GCAACTGCGT GCACTTCGTG GCCGAGGAGC AGGACTGACC GACGCCGACC 2281 AACACCGCCG GTCCGACGGC GGCCCACGGG TCCCAGGCCT CGGAGATCCT AACACCGGGT 2341 TGTGTGGCCA AAATTGTTCT GTAGTCGCTG TGAGTTGACA CGGCTAGTGC TTATGATTTT 2401 CCTCGCGTGT GGTGCCTGTA CTCAGCCCTA TGCCTTATTT GCAACACATT TACGTACAGC 2461 GCACAAGAGA AGAGAAGATC ACTTGAAGAT AATAAATATA GGGTTGTAGG CATCTTGTTT 2521 AACTCAAATT TTCTCGTCTT GGTGTGTCGA CATGATTGAA ATAGTGCCAC CAGTTGTGTT 2581 TGATGCGTTT GTTATCTATG CAGTATTCTG CAGGCTGCTA ACAAAGCCCG AAAGGAAGCT 2641 GAGTTGGCTG CTGCCACCGC TGAGCAATAA CTAGCATAAC CCCTTGGGGC CTCTAAACGG 2701 GTCTTGAGGG GTTTTTTGCT GAAAGGAGGA ACTATATCCG GATAGGGCTG CTAACAAAGC 2761 CCGAAAGGAA GCTGAGTTGG CTGCTGCCAC CGCTGAGCAA TAACTAGCAT AACCCCTTGG 2821 GGCCTCTAAA CGGGTCTTGA GGGGTTTTTT GCTGAAAGGA GGAACTATAT CCGGATAGGG 2881 CCCCTGCAGG CATGCAAGCT AGCCAGATCC TCTACGCCGG ACGCATCGTG GCCGGCATCA 2941 CCGGCGCCAC AGGTGCGGTT GCTGGCGCCT ATATCGCCGA CATCACCGAT GGGGAAGATC 3001 GGGCTCGCCA CTTCGGGCTC ATGAGCGCTT GTTTCGGCGT GGGTATGGTG GCAGGCCCCG 3061 TGGCCGGGGG ACTGTTGGGC GCCATCTCCT TGCACCATTC CTTGCGGCGG CGGTGCTCAA 3121 CGGCCTCAAC CTACTACTGG GCTGCTTCCT AATGCAGGAG TCGCATAAGG GAGAGCGTCG 3181 ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT AGTTAAGCCA GCCCCGACAC 3241 CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA 3301 CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA 3361 CGCGCGAGAC GAAAGGGCCT CGTGATACGC CTATTTTTAT AGGTTAATGT CATGATAATA 3421 ATGGTTTCTT AGACGTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC CCCTATTTGT 3481 TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA GACAATAACC CTGATAAATG 3541 CTTCAATAAT ATTGAAAAAG GAAGAGTATG AGTATTCAAC ATTTCCGTGT CGCCCTTATT 3601 CCCTTTTTTG CGGCATTTTG CCTTCCTGTT TTTGCTCACC CAGAAACGCT GGTGAAAGTA 3661 AAAGATGCTG AAGATCAGTT GGGTGCACGA GTGGGTTACA TCGAACTGGA TCTCAACAGC 3721 GGTAAGATCC TTGAGAGTTT TCGCCCCGAA GAACGTTTTC CAATGATGAG CACTTTTAAA 3781 GTTCTGCTAT GTGGCGCGGT ATTATCCCGT ATTGACGCCG GGCAAGAGCA ACTCGGTCGC 3841 CGCATACACT ATTCTCAGAA TGACTTGGTT GAGTACTCAC CAGTCACAGA AAAGCATCTT 3901 ACGGATGGCA TGACAGTAAG AGAATTATGC AGTGCTGCCA TAACCATGAG TGATAACACT 3961 GCGGCCAACT TACTTCTGAC AACGATCGGA GGACCGAAGG AGCTAACCGC TTTTTTGCAC 4021 AACATGGGGG ATCATGTAAC TCGCCTTGAT CGTTGGGAAC CGGAGCTGAA TGAAGCCATA 4081 CCAAACGACG AGCGTGACAC CACGATGCCT GTAGCAATGG CAACAACGTT GCGCAAACTA 4141 TTAACTGGCG AACTACTTAC TCTAGCTTCC CGGCAACAAT TAATAGACTG GATGGAGGCG 4201 GATAAAGTTG CAGGACCACT TCTGCGCTCG GCCCTTCCGG CTGGCTGGTT TATTGCTGAT 4261 AAATCTGGAG CCGGTGAGCG TGGGTCTCGC GGTATCATTG CAGCACTGGG GCCAGATGGT 4321 AAGCCCTCCC GTATCGTAGT TATCTACACG ACGGGGAGTC AGGCAACTAT GGATGAACGA 4381 AATAGACAGA TCGCTGAGAT AGGTGCCTCA CTGATTAAGC ATTGGTAACT GTCAGACCAA 4441 GTTTACTCAT ATATACTTTA GATTGATTTA AAACTTCATT TTTAATTTAA AAGGATCTAG 4501 GTGAAGATCC TTTTTGATAA TCTCATGACC AAAATCCCTT AACGTGAGTT TTCGTTCCAC 4561 TGAGCGTCAG ACCCCGTAGA AAAGATCAAA GGATCTTCTT GAGATCCTTT TTTTCTGCGC 4621 GTAATCTGCT GCTTGCAAAC AAAAAAACCA CCGCTACCAG CGGTGGTTTG TTTGCCGGAT 4681 CAAGAGCTAC CAACTCTTTT TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA GATACCAAAT 4741 ACTGTCCTTC TAGTGTAGCC GTAGTTAGGC CACCACTTCA AGAACTCTGT AGCACCGCCT 4801 ACATACCTCG CTCTGCTAAT CCTGTTACCA GTGGCTGCTG CCAGTGGCGA TAAGTCGTGT 4861 CTTACCGGGT TGGACTCAAG ACGATAGTTA CCGGATAAGG CGCAGCGGTC GGGCTGAACG 4921 GGGGGTTCGT GCACACAGCC CAGCTTGGAG CGAACGACCT ACACCGAACT GAGATACCTA 4981 CAGCGTGAGC TATGAGAAAG CGCCACGCTT CCCGAAGGGA GAAAGGCGGA CAGGTATCCG 5041 GTAAGCGGCA GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG AAACGCCTGG 5101 TATCTTTATA GTCCTGTCGG GTTTCGCCAC CTCTGACTTG AGCGTCGATT TTTGTGATGC 5161 TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC GCCAGCAACG CGGCCTTTTT ACGGTTCCTG 5221 GCCTTTTGCT GGCCTTTTGC TCACATGTTC TTTCCTGCGT TATCCCCTGA TTCTGTGGAT 5281 AACCGTATTA CCGCCTTTGA GTGAGCTGAT ACCGCTCGCC GCAGCCGAAC GACCGAGCGC 5341 AGCGAGTCAG TGAGCGAGGA AGCGGAAGAG CGCCCAATAC GCAAACCGCC TCTCCCCGCG 5401 CGTTGGCCGA TTCATTAATG CAG //