LOCUS pUB39 6498 bp DNA circular 25-JUL-2008 Made bu Ulrike Boehme. Sequence reconstructed 20080723 by GAMC. The length of the plasmid differs (is 179 more than) that in Uli's original GCK file. The difference appears to be due to a deletion in the GCK file of 179 bp of vector sequence downstream of the T7 terminators. I assume this was an inadvertent error at some time. The current version is, I assume, the definitive (albeit predicted) sequence. FEATURES Location/Qualifiers misc_feature complement(1..713) /function="rRNA Spacer" promoter 720..739 /function="T7 Promoter" /note="T7 promoter & initiation region. Promoter-Op-Luc cassette orig from Lew19" 5'UTR 770..825 /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 826..856 /function="GPEET 5´ UTR" 3'UTR 2699..2845 /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 2845..2959 /function="ACT 5´ UTR" /note="actin-derived sas; used in all HD330, 430 & Lew5 descendants" CDS 2969..3340 /function="BLE" 3'UTR 3395..3682 /function="ACT 3´ UTR" /note="actin 3'UTR- same as what's used in all HD330, 430 and Lew5 descendants" terminator 3689..3821 /function="T7 Term" /note="132bp Pet3a fragment (originally PCR'd), then subcloned from published HD216, bearing T7 terminator" terminator 3822..3954 /function="T7 Term" /note="132 bp Pet3a fragment (originally PCR'd then subcloned from published HD216) bearing T7 terminator" CDS 4643..5500 /function="LACTAMASE" conflict 766..773 /note="From a restriction digest of a pLew111 derivative, an SrfI site (GCCCGGGC) originally designated here was absent, according to Shawn Motyka (Barbara Sollner Webb lab) email 9/8/3. This was confirmed by George from 2 seq trace files of Fabian's pLew111-bsd construct." 5'UTR 860..976 /function="VSG 5' UTR" CDS 980..2561 /function="VSG 117" 3'UTR 2561..2676 /note="VSG 117 3' UTR" misc_feature 2612..2627 /function="Conserved 16-mer" protein_bind 745..763 /function="Tet Operator" BASE COUNT 1803 a 1606 c 1672 g 1417 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 AGATCTCCCG GGCTGCACGC 781 GCCTTCGAGT TTTTTTTCCT TTTCCCCATT TTTTTCAACT TGAAGACTTC AATTACACCA 841 AAAAGTAAAA TTCACAAGCT aattccagct gtcgacgcgg ggtttctgta ctatattgca 901 gcaaaagact agaagcaagc agcgcatata gcgcaaacaa tcggggtttc aacaaaaacg 961 ggagcgactc acaAAGCTTa tggactgcca tacaaaggag acactagggg tcacacaatg 1021 gaggcgatca acgatgctaa cactatcact gctttacgcc atcactccag cggacggcgc 1081 caaagaagcc cttgaataca aaacttggac aaaccactgc ggactggcgg ccacactgag 1141 aaaggttgcc ggtggagtat taacgaaact gaaaagccac attagctacc ggaaaaaact 1201 ggaagaaatg gaaacgaagc tacgaatcta cgcactaaaa ggagacggag tgggagagca 1261 aaaatcagcg gagatactag caacaacggc agccctaatg cgacaaaaag cactcacacc 1321 agaagaagca aatttgaaaa cagcgctgaa ggcggcagga ttcgcaggcg aaggagcggc 1381 agccgtcagc agctacctga tgacactcgg gacactgaca acaagcggat ctgcgcactg 1441 cctaagcaac gaaggcggcg acggtgacgg aaaagaccaa cttgcgccga aaggctgccg 1501 gcacggcaca gaagcagact tcgacgcagg agccggcccg gcagaatctg aagtagccga 1561 cagcggcttc gcgcaagtac caggcaaaca ggacggagca aacgcaggcc aagcaaacat 1621 gtgcgcattg ttcacacacc aagcaacgcc gcacagctca cagggcatat acataaccgg 1681 ggcacagaca aaaccttcat tcgggtacgg catgctgaca atcggcacga cggaccagac 1741 catcggcttg aaactttcgg acataaaggg caaacaagca gacagcgcgc agaaattctg 1801 gagcagctgc cacgcagcag tcaaagccgc ccaagatatg aaggcagacc cagccctaaa 1861 agtcgaccag acgctcctag ctgttcttgt ggcttctccg gagatggctg aaatactgaa 1921 actagaagcg gcagcatcac agcaaaaagg accagaggaa gtgacgatcg acctagccac 1981 cgagaaaaac aattatttcg gaaccaacaa caacaaacta gagccgctct ggactaaaat 2041 caaaggacag aatatagttg acttggcggc gaccaaaggc agcacgaaag agttaggaac 2101 agtcacagac acggccgagc tacaaaaact tttaagttat tattacacgg tcaacaaaga 2161 agaacagaaa aaaacagcgg agaaaataac taaactcgaa accgaactag cagatcaaaa 2221 aggcaaatcc cctgaaagcg agtgcaataa aatatctgag gaacccaaat gcaacgagga 2281 caagatatgc agttggcata aggaggttaa agcgggagaa aagcactgca aatttaactc 2341 aacaaaagca aaagaaaagg gggtctctgt aacacaaact caaactgcag gaggaaccga 2401 agcgacaaca gataaatgca aagggaaatt ggaagatacc tgcaagaagg agagcaactg 2461 caaatgggaa aataatgctt gcaaagattc ctctattcta gtaaccaaga aattcgccct 2521 caccgtggtt tctgctgcat ttgtggcctt gcttttttaa GGATCCtttt ccccctcttt 2581 ttcttaaaaa ttcttgctac ttgaaaactc ctgatatatt ttaacacgca aattacccga 2641 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaacccg cgtcgacagc tggaattGCC 2701 CATTTAGTTG GCTTTTCCCT TGTCTCGTGT CTTTTCCGTG GAAAGGTTCC CGGAGTAATC 2761 TGATGGCACA GCAGGGAGGT GCGCCTGCAG GTTGGTTAGG AAGGGGGGAT GATGTAAAAG 2821 AAGAAAATGG GGGGATTCGA GCCCGGGCAC AGCAAGGTCT TCTGAAATTC ATGTTTTTTT 2881 TTTTTTTACT CTGCATTGCA GTCTCCGCTC TTATTTAGTT TTGCTTTACG TAAGGTCTCG 2941 TTGCTGCCAT AAAATAAGCT ATATCACCAT GGCCAAGTTG ACCAGTGCCG TTCCGGTGCT 3001 CACCGCGCGC GACGTCGCCG GAGCGGTCGA GTTCTGGACC GACCGGCTCG GGTTCTCCCG 3061 GGACTTCGTG GAGGACGACT TCGCCGGTGT GGTCCGGGAC GACGTGACCC TGTTCATCAG 3121 CGCGGTCCAG GACCAGGTGG TGCCGGACAA CACCCTGGCC TGGGTGTGGG TGCGCGGCCT 3181 GGACGAGCTG TACGCCGAGT GGTCGGAGGT CGTGTCCACG AACTTCCGGG ACGCCTCCGG 3241 GCCGGCCATG ACCGAGATCG GCGAGCAGCC GTGGGGGCGG GAGTTCGCCC TGCGCGACCC 3301 GGCCGGCAAC TGCGTGCACT TCGTGGCCGA GGAGCAGGAC TGACCGACGC CGACCAACAC 3361 CGCCGGTCCG ACGGCGGCCC ACGGGTCCCA GGCCTCGGAG ATCCTAACAC CGGGTTGTGT 3421 GGCCAAAATT GTTCTGTAGT CGCTGTGAGT TGACACGGCT AGTGCTTATG ATTTTCCTCG 3481 CGTGTGGTGC CTGTACTCAG CCCTATGCCT TATTTGCAAC ACATTTACGT ACAGCGCACA 3541 AGAGAAGAGA AGATCACTTG AAGATAATAA ATATAGGGTT GTAGGCATCT TGTTTAACTC 3601 AAATTTTCTC GTCTTGGTGT GTCGACATGA TTGAAATAGT GCCACCAGTT GTGTTTGATG 3661 CGTTTGTTAT CTATGCAGTA TTCTGCAGGC TGCTAACAAA GCCCGAAAGG AAGCTGAGTT 3721 GGCTGCTGCC ACCGCTGAGC AATAACTAGC ATAACCCCTT GGGGCCTCTA AACGGGTCTT 3781 GAGGGGTTTT TTGCTGAAAG GAGGAACTAT ATCCGGATAG GGCTGCTAAC AAAGCCCGAA 3841 AGGAAGCTGA GTTGGCTGCT GCCACCGCTG AGCAATAACT AGCATAACCC CTTGGGGCCT 3901 CTAAACGGGT CTTGAGGGGT TTTTTGCTGA AAGGAGGAAC TATATCCGGA TAGGGCCCCT 3961 GCAGGCATGC AAGCTAGCCA GATCCTCTAC GCCGGACGCA TCGTGGCCGG CATCACCGGC 4021 GCCACAGGTG CGGTTGCTGG CGCCTATATC GCCGACATCA CCGATGGGGA AGATCGGGCT 4081 CGCCACTTCG GGCTCATGAG CGCTTGTTTC GGCGTGGGTA TGGTGGCAGG CCCCGTGGCC 4141 GGGGGACTGT TGGGCGCCAT CTCCTTGCAC CATTCCTTGC GGCGGCGGTG CTCAACGGCC 4201 TCAACCTACT ACTGGGCTGC TTCCTAATGC AGGAGTCGCA TAAGGGAGAG CGTCGATATG 4261 GTGCACTCTC AGTACAATCT GCTCTGATGC CGCATAGTTA AGCCAGCCCC GACACCCGCC 4321 AACACCCGCT GACGCGCCCT GACGGGCTTG TCTGCTCCCG GCATCCGCTT ACAGACAAGC 4381 TGTGACCGTC TCCGGGAGCT GCATGTGTCA GAGGTTTTCA CCGTCATCAC CGAAACGCGC 4441 GAGACGAAAG GGCCTCGTGA TACGCCTATT TTTATAGGTT AATGTCATGA TAATAATGGT 4501 TTCTTAGACG TCAGGTGGCA CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT 4561 TTTCTAAATA CATTCAAATA TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA 4621 ATAATATTGA AAAAGGAAGA GTATGAGTAT TCAACATTTC CGTGTCGCCC TTATTCCCTT 4681 TTTTGCGGCA TTTTGCCTTC CTGTTTTTGC TCACCCAGAA ACGCTGGTGA AAGTAAAAGA 4741 TGCTGAAGAT CAGTTGGGTG CACGAGTGGG TTACATCGAA CTGGATCTCA ACAGCGGTAA 4801 GATCCTTGAG AGTTTTCGCC CCGAAGAACG TTTTCCAATG ATGAGCACTT TTAAAGTTCT 4861 GCTATGTGGC GCGGTATTAT CCCGTATTGA CGCCGGGCAA GAGCAACTCG GTCGCCGCAT 4921 ACACTATTCT CAGAATGACT TGGTTGAGTA CTCACCAGTC ACAGAAAAGC ATCTTACGGA 4981 TGGCATGACA GTAAGAGAAT TATGCAGTGC TGCCATAACC ATGAGTGATA ACACTGCGGC 5041 CAACTTACTT CTGACAACGA TCGGAGGACC GAAGGAGCTA ACCGCTTTTT TGCACAACAT 5101 GGGGGATCAT GTAACTCGCC TTGATCGTTG GGAACCGGAG CTGAATGAAG CCATACCAAA 5161 CGACGAGCGT GACACCACGA TGCCTGTAGC AATGGCAACA ACGTTGCGCA AACTATTAAC 5221 TGGCGAACTA CTTACTCTAG CTTCCCGGCA ACAATTAATA GACTGGATGG AGGCGGATAA 5281 AGTTGCAGGA CCACTTCTGC GCTCGGCCCT TCCGGCTGGC TGGTTTATTG CTGATAAATC 5341 TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT CATTGCAGCA CTGGGGCCAG ATGGTAAGCC 5401 CTCCCGTATC GTAGTTATCT ACACGACGGG GAGTCAGGCA ACTATGGATG AACGAAATAG 5461 ACAGATCGCT GAGATAGGTG CCTCACTGAT TAAGCATTGG TAACTGTCAG ACCAAGTTTA 5521 CTCATATATA CTTTAGATTG ATTTAAAACT TCATTTTTAA TTTAAAAGGA TCTAGGTGAA 5581 GATCCTTTTT GATAATCTCA TGACCAAAAT CCCTTAACGT GAGTTTTCGT TCCACTGAGC 5641 GTCAGACCCC GTAGAAAAGA TCAAAGGATC TTCTTGAGAT CCTTTTTTTC TGCGCGTAAT 5701 CTGCTGCTTG CAAACAAAAA AACCACCGCT ACCAGCGGTG GTTTGTTTGC CGGATCAAGA 5761 GCTACCAACT CTTTTTCCGA AGGTAACTGG CTTCAGCAGA GCGCAGATAC CAAATACTGT 5821 CCTTCTAGTG TAGCCGTAGT TAGGCCACCA CTTCAAGAAC TCTGTAGCAC CGCCTACATA 5881 CCTCGCTCTG CTAATCCTGT TACCAGTGGC TGCTGCCAGT GGCGATAAGT CGTGTCTTAC 5941 CGGGTTGGAC TCAAGACGAT AGTTACCGGA TAAGGCGCAG CGGTCGGGCT GAACGGGGGG 6001 TTCGTGCACA CAGCCCAGCT TGGAGCGAAC GACCTACACC GAACTGAGAT ACCTACAGCG 6061 TGAGCTATGA GAAAGCGCCA CGCTTCCCGA AGGGAGAAAG GCGGACAGGT ATCCGGTAAG 6121 CGGCAGGGTC GGAACAGGAG AGCGCACGAG GGAGCTTCCA GGGGGAAACG CCTGGTATCT 6181 TTATAGTCCT GTCGGGTTTC GCCACCTCTG ACTTGAGCGT CGATTTTTGT GATGCTCGTC 6241 AGGGGGGCGG AGCCTATGGA AAAACGCCAG CAACGCGGCC TTTTTACGGT TCCTGGCCTT 6301 TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC CCTGATTCTG TGGATAACCG 6361 TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC CGAACGACCG AGCGCAGCGA 6421 GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA CCGCCTCTCC CCGCGCGTTG 6481 GCCGATTCAT TAATGCAG //