LOCUS pLEW100v5b1d-HYG 8284 bp DNA circular 18-JUL-2008 FEATURES Location/Qualifiers misc_feature complement(23..714) /function="rRNA spacer" /note="Complement of 36 to 694 of accession number Z46903 and complement of 809632 to 810308 on chromosome 1 (AL929603) (the entire 'intergenic region' on chromosome 1 is about 10 kb." terminator complement(730..861) /function="T7 Terminator" /note="Actual sequence 9/26/5" terminator complement(862..993) /function="T7 Terminator" /note="Actual sequence 9/26/5" 3'UTR complement(1002..1289) /function="ACT 3´ UTR" /note="When the entire downstream region was BLASTed, these are the nt that matched the 427 Bn-2 ES. The match was almost the same against the LiTat1.6 ACT, with 3 internal point differences" CDS complement(1304..2326) /function="HYG" conflict 2334..2338 /function="" /note="Sequence correction Jan 2004: XbaI site created by a 5bp insertion" 5'UTR complement(2339..2454) /function="ACT UTR & upstream" promoter complement(2458..2477) /function="T7 Promoter" /note="T7 promoter & initiation region. Promoter-Op-Luc cassette orig from Lew19" promoter 2489..2747 /function="rRNA Promoter" /note="395 to 655 in AF416290 and repeated in several chromosomes, for example ChrI AL929603 792760 to 793004" protein_bind 2752..2792 /function="2 Tet Op" 5'UTR 2796..2916 /function="GPEET 5' UTR & upstream" CDS 2987..4636 /function="LUCIFERASE" 3'UTR 4790..5501 /function="ALD 3´ long UTR" /note="long aldolase 3' UTR from pHD103 for optimized BF expression (about 6x higher than the short ALD UTR used in pLew79 & pLew82)." /note="Note that there are no T7 terminators downstream of T7 transcription unit in this construct, which could be a problem in some insertion sites!" CDS 6430..7287 /function="LACTAMASE" misc_feature 4640..4789 /note="Extraneous vector sequence" misc_signal 2747..2747 /function="Transcription initiation" /note="According to White Rudenko & Borst 1986, explored in Janz & Clayton 1994 and Vanhamme et al 1995 and reviewed in Hotz & clayton 1998, mutations in the A or the preceeding T reduced transcription 10-50x." conflict 2751..2751 /note="Changed one nt to remove the Bam site. See ina email january 5 2007 for logic of choosing the nt to change (the promoter and the operator sequences are very close)." CDS 2944..2985 /function="Extraneous ORF" BASE COUNT 2199 a 1937 c 2034 g 2114 t ORIGIN 1 GAATTCGAGC TCCAGCTGAG CTCATATAGT TGGTATGTAT TCTAATTCCA GACTACTGGC 61 GTGGATAAAC ATGTCCCCTG ATTAAAAGGA AAGATTCCAT AGCCCATTAG TGCAAAGATA 121 ATTGGTACAC CGTCAAAAAC ACATGGGAGA CCACACGATA CGACACTGTG ACCGTAGCAT 181 CAATTCCGCA CTCGATATAA CTCTATCGAA GAACTTCCAT GGTACAAACT GGTCGCCACG 241 GTCTGCTCCA CACGGAGATC ATCGTATCAT TTTTATCGAT AGCGGCCGCT ATCGATGTAT 301 GCCTTGGCCC TGATGGCATG CCAATTTCAC TACAACGGAC TATGTGGACC CCGTTATCAT 361 GGAAATGCGC TAGTTGGAGG AAGTTAGACC GCGCCGGAAA AGAGAGGGGT AGAGAAAATG 421 ACAACTTGGA AGATATCCAC ACACGCACGG TGAAACGTTA GCAACAATTA TTAGGGAAGC 481 ACGCTTGCCC TAGTCCCACG AGTAACCAAG ACTCCAAAAG CCTTTCTGGC ACAGAGAGCG 541 AGCCGAAATG GAAAAGAGAA ACAATGCCTG CACTAACACT ACTGAGCGAT TCGCCTCGCC 601 GCGGAGGACC GAATACTAAT AACGACACTT GCGGTCAAAA AGTAGAAGAA CAAATGCTCA 661 ACGATGAGTG AATCAGGTTA GGGTAGTTGG AAAATTATAC AGAATGTCTT TGGCAACACA 721 CCGCTTAAGC CTATCCGGAT ATAGTTCTCC TTTCAGCAAA AAACCCCTCA AGACCCGTTT 781 AGAGGCCCCA AGGGGTTATG CTAGTTATTG CTCAGCGGTG GCAGCAGCCA ACTCAGCTTC 841 CTTTCGGGCT TTGTTAGCAG CCCTATCCGG ATATAGTTCT CCTTTCAGCA AAAAACCCCT 901 CAAGACCCGT TTAGAGGCCC CAAGGGGTTA TGCTAGTTAT TGCTCAGCGG TGGCAGCAGC 961 CAACTCAGCT TCCTTTCGGG CTTTGTTAGC AGCATTTAAA TTTGCAGAAT ACTGCATAGA 1021 TAACAAACGC ATCAAACACA ACTGGTGGCA CTATTTCAAT CATGTCGACA CACCAAGACG 1081 AGAAAATTTG AGTTAAACAA GATGCCTACA ACCCTATATT TATTATCTTC AAGTGATCTT 1141 CTCTTCTCTT GTGCGCTGTA CGTAAATGTG TTGCAAATAA GGCATAGGGC TGAGTACAGG 1201 CACCACACGC GAGGAAAATC ATAAGCACTA GCCGTGTCAA CTCACAGCGA CTACAGAACA 1261 ATTTTGGCCA CACAACCCGG TGTTAGGATC TCCGAGGCCT CTATTCCTTT GCCCTCGGAC 1321 GAGTGCTGGG GCGTCGGTTT CCACTATCGG CGAGTACTTC TACACAGCCA TCGGTCCAGA 1381 CGGCCGCGCT TCTGCGGGCG ATTTGTGTAC GCCCGACAGT CCCGGCTCCG GATCGGACGA 1441 TTGCGTCGCA TCGACCCTGC GCCCAAGCTG CATCATCGAA ATTGCCGTCA ACCAAGCTCT 1501 GATAGAGTTG GTCAAGACCA ATGCGGAGCA TATACGCCCG GAGCCGCGGC GATCCTGCAA 1561 GCTCCGGATG CCTCCGCTCG AAGTAGCGCG TCTGCTGCTC CATACAAGCC AACCACGGCC 1621 TCCAGAAGAA GATGTTGGCG ACCTCGTATT GGGAATCCCC GAACATCGCC TCGCTCCAGT 1681 CAATGACCGC TGTTATGCGG CCATTGTCCG TCAGGACATT GTTGGAGCCG AAATCCGCGT 1741 GCACGAGGTG CCGGACTTCG GGGCAGTCCT CGGCCCAAAG CATCAGCTCA TCGAGAGCCT 1801 GCGCGACGGA CGCACTGACG GTGTCGTCCA TCACAGTTTG CCAGTGATAC ACATGGGGAT 1861 CAGCAATCGC GCATATGAAA TCACGCCATG TAGTGTATTG ACCGATTCCT TGCGGTCCGA 1921 ATGGGCCGAA CCCGCTCGTC TGGCTAAGAT CGGCCGCAGC GATCGCATCC ATGGCCTCCG 1981 CGACCGGCTG CAGAACAGCG GGCAGTTCGG TTTCAGGCAG GTCTTGCAAC GTGACACCCT 2041 GTGCACGGCG GGAGATGCAA TAGGTCAGGC TCTCGCTGAA TTCCCCAATG TCAAGCACTT 2101 CCGGAATCGG GAGCGCGGCC GATGCAAAGT GCCGATAAAC ATAACGATCT TTGTAGAAAC 2161 CATCGGCGCA GCTATTTACC CGCAGGACAT ATCCACGCCC TCCTACATCG AAGCTGAAAG 2221 CACGAGATTC TTCGCCCTCC GAGAGCTGCA TCAGGTCGGA GACGCTGTCG AACTTTTCGA 2281 TCAGAAACTT CTCGACAGAC GTCGCGGTGA GTTCAGGCTT TTTCATCACT AGTTCTAGAG 2341 CTTATTTTAT GGCAGCAACG AGACCTTACG TAAAGCAAAA CTAAATAAGA GCGGAGACTG 2401 CAATGCAGAG TAAAAAAAAA AAAAACATGA ATTTCAGAAG ACCTTGCTGT GCCCCCTCCC 2461 TATAGTGAGT CGTATTAATG GTACCTAGCT TTCCACCCAG CGCGGGTGCA TTCTGGCTCT 2521 TATATATACT TATTGTCATG ACAGAGTATA TTGTACTGTG TTGATAAGGG ACGGGTAACT 2581 GTATTGAAGA GCCGATGCTT TTGACATGTT AGATATAATA TGTTTTATTG TAAAGTCAAT 2641 ACAACACACA ATAGGATAAT AATGATAAAG TTAAAAAAGT ATATATAGTA ATAGAAATAT 2701 ATCTTATATA GGAAAGATTA AGCAGTAAAA GTAGCGCTTA CGGCGTACGG TTCCCTATCA 2761 GTGATAGAGA TCTCCCTATC AGTGATAGAG ATCCCTGAGT ACTGAGTTTA ACATGTTCTC 2821 GTCCCGGGCT GCACGCGCCT TCGAGTTTTT TTTCCTTTTC CCCATTTTTT TCAACTTGAA 2881 GACTTCAATT ACACCAAAAA GTAAAATTCA CAAGCTTGGA ATTCCTTTGT GTTACATTCT 2941 TGAATGTCGC TCGCAGTGAC ATTAGCATTC CGGTACTGTT GGTAAAATGG AAGACGCCAA 3001 AAACATAAAG AAAGGCCCGG CGCCATTCTA TCCTCTAGAG GATGGAACCG CTGGAGAGCA 3061 ACTGCATAAG GCTATGAAGA GATACGCCCT GGTTCCTGGA ACAATTGCTT TTACAGATGC 3121 ACATATCGAG GTGAACATCA CGTACGCGGA ATACTTCGAA ATGTCCGTTC GGTTGGCAGA 3181 AGCTATGAAA CGATATGGGC TGAATACAAA TCACAGAATC GTCGTATGCA GTGAAAACTC 3241 TCTTCAATTC TTTATGCCGG TGTTGGGCGC GTTATTTATC GGAGTTGCAG TTGCGCCCGC 3301 GAACGACATT TATAATGAAC GTGAATTGCT CAACAGTATG AACATTTCGC AGCCTACCGT 3361 AGTGTTTGTT TCCAAAAAGG GGTTGCAAAA AATTTTGAAC GTGCAAAAAA AATTACCAAT 3421 AATCCAGAAA ATTATTATCA TGGATTCTAA AACGGATTAC CAGGGATTTC AGTCGATGTA 3481 CACGTTCGTC ACATCTCATC TACCTCCCGG TTTTAATGAA TACGATTTTG TACCAGAGTC 3541 CTTTGATCGT GACAAAACAA TTGCACTGAT AATGAATTCC TCTGGATCTA CTGGGTTACC 3601 TAAGGGTGTG GCCCTTCCGC ATAGAACTGC CTGCGTCAGA TTCTCGCATG CCAGAGATCC 3661 TATTTTTGGC AATCAAATCA TTCCGGATAC TGCGATTTTA AGTGTTGTTC CATTCCATCA 3721 CGGTTTTGGA ATGTTTACTA CACTCGGATA TTTGATATGT GGATTTCGAG TCGTCTTAAT 3781 GTATAGATTT GAAGAAGAGC TGTTTTTACG ATCCCTTCAG GATTACAAAA TTCAAAGTGC 3841 GTTGCTAGTA CCAACCCTAT TTTCATTCTT CGCCAAAAGC ACTCTGATTG ACAAATACGA 3901 TTTATCTAAT TTACACGAAA TTGCTTCTGG GGGCGCACCT CTTTCGAAAG AAGTCGGGGA 3961 AGCGGTTGCA AAACGCTTCC ATCTTCCAGG GATACGACAA GGATATGGGC TCACTGAGAC 4021 TACATCAGCT ATTCTGATTA CACCCGAGGG GGATGATAAA CCGGGCGCGG TCGGTAAAGT 4081 TGTTCCATTT TTTGAAGCGA AGGTTGTGGA TCTGGATACC GGGAAAACGC TGGGCGTTAA 4141 TCAGAGAGGC GAATTATGTG TCAGAGGACC TATGATTATG TCCGGTTATG TAAACAATCC 4201 GGAAGCGACC AACGCCTTGA TTGACAAGGA TGGATGGCTA CATTCTGGAG ACATAGCTTA 4261 CTGGGACGAA GACGAACACT TCTTCATAGT TGACCGCTTG AAGTCTTTAA TTAAATACAA 4321 AGGATATCAG GTGGCCCCCG CTGAATTGGA ATCGATATTG TTACAACACC CCAACATCTT 4381 CGACGCGGGC GTGGCAGGTC TTCCCGACGA TGACGCCGGT GAACTTCCCG CCGCCGTTGT 4441 TGTTTTGGAG CACGGAAAGA CGATGACGGA AAAAGAGATC GTGGATTACG TCGCCAGTCA 4501 AGTAACAACC GCGAAAAGGT TGCGCGGAGG AGTTGTGTTT GTGGACGAAG TACCGAAAGG 4561 TCTTACCGGA AAACTCGACG CAAGAAAAAT CAGAGAGATC CTCATAAAGG CCAAGAAGGG 4621 CGGAAAGTCC AAATTGTAAA ATGTAACTGT ATTCAGCGAT GACGAAATTC TTAGCTATTG 4681 TAATATTATA TGCAAATTGA TGAATGGTAA TTTTGTAATT GTGGGTCACT GTACTATTTT 4741 AACGAATAAT AAAATCAGGT ATAGGTAACT AAAAAGGAAT TCGAGCTCGG ATCCTGCCCA 4801 TTTAGTTGGC TTTTCCCTTG TCTCGTGTCT TTTCCGTGGA AAGGTTCCCG GAGTAATCTG 4861 ATGGCACAGC AGGGAGGTGC GCCTGCAGGT TGGTTAGGAA GGGGGGATGA TGTAAAAGAA 4921 GAAAATGGGG GGATATCTTA TGTTTAAGAG GATAAATAAT GTGAAGGGGC TTTATATGCT 4981 TGCTTGGTTG TCTCGTTGGT GCATGGGGAA TCTGCATGTT TGCTTTGGAG CACGCGTGGT 5041 ACATCAATGG GATCATATCT TGCTGCCTCC CGCAGTCACC GTGCGGAGCT GCCGGTGCCC 5101 CGTTTCTCAT AGGCGTGTGC ACCTTCCGTC TGAGACCTGT AAATGGTTAA AAAGGAATAT 5161 ATAATTACCT TTTGAAAAGT GGTAAAAAAC GAATATATGT TTTTTTTTGG TTTTAATACG 5221 CTCTTTTGTG TGTATGAGAG GAATAAATAA GTGTGTGTGT GTGTGTGTGT TGTTGTTGTT 5281 GAAAAAATGA AGGACACGAA AGCGCTCATT AGGGGTGGGG ATACACCAGG CATAATTATC 5341 CGAAAATATT TTGGTGCACC AAATAAGTGA ACATACTGAC GAAATCAAAG AGTTGGAGGA 5401 TGGATAGGGA GGCCTCATTG GCAGTGGTAA ATTGATTTGA CTTGAATATG ATACCCTCTT 5461 ACTGTTTCAT TGCCACTACA AGTTGGTTTC CTTCCCCTGC AGGCATGCAA GCTAGCTTGT 5521 ATTCTATAGT GTCACCTAAA TCGTATGTGT ATGATACATA AGGTTATGTA TTAATTGTAG 5581 CCGCGTTCTA ACGACAATAT GTACAAGCCT AATTGTGTAG CATCTGGCTT ACTGAAGCAG 5641 ACCCTATCAT CTCTCTCGTA AACTGCCGTC AGAGTCGGTT TGGTTGGACG AACCTTCTGA 5701 GTTTCTGGTA ACGCCGTCCC GCACCCGGAA ATGGTCAGCG AACCAATCAG CAGGGTCATC 5761 GCTAGCCAGA TCCTCTACGC CGGACGCATC GTGGCCGGCA TCACCGGCGC CACAGGTGCG 5821 GTTGCTGGCG CCTATATCGC CGACATCACC GATGGGGAAG ATCGGGCTCG CCACTTCGGG 5881 CTCATGAGCG CTTGTTTCGG CGTGGGTATG GTGGCAGGCC CCGTGGCCGG GGGACTGTTG 5941 GGCGCCATCT CCTTGCACCA TTCCTTGCGG CGGCGGTGCT CAACGGCCTC AACCTACTAC 6001 TGGGCTGCTT CCTAATGCAG GAGTCGCATA AGGGAGAGCG TCGAATGGTG CACTCTCAGT 6061 ACAATCTGCT CTGATGCCGC ATAGTTAAGC CAGCCCCGAC ACCCGCCAAC ACCCGCTGAC 6121 GCGCCCTGAC GGGCTTGTCT GCTCCCGGCA TCCGCTTACA GACAAGCTGT GACCGTCTCC 6181 GGGAGCTGCA TGTGTCAGAG GTTTTCACCG TCATCACCGA AACGCGCGAG ACGAAAGGGC 6241 CTCGTGATAC GCCTATTTTT ATAGGTTAAT GTCATGATAA TAATGGTTTC TTAGACGTCA 6301 GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT CTAAATACAT 6361 TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA ATATTGAAAA 6421 AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT 6481 TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG 6541 TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT CCTTGAGAGT 6601 TTTCGCCCCG AAGAACGTTT TCCAATGATG AGCACTTTTA AAGTTCTGCT ATGTGGCGCG 6661 GTATTATCCC GTATTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA CTATTCTCAG 6721 AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA 6781 AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG 6841 ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAGCATGGG GGATCATGTA 6901 ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA CGAGCGTGAC 6961 ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG CGAACTACTT 7021 ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA 7081 CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG 7141 CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC CCGTATCGTA 7201 GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA GATCGCTGAG 7261 ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC ATATATACTT 7321 TAGATTGATT TAAAACTTCA TTTTTAATTT AAAAGGATCT AGGTGAAGAT CCTTTTTGAT 7381 AATCTCATGA CCAAAATCCC TTAACGTGAG TTTTCGTTCC ACTGAGCGTC AGACCCCGTA 7441 GAAAAGATCA AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA 7501 ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT 7561 TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTTCT TCTAGTGTAG 7621 CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA 7681 ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA 7741 AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG 7801 CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA GCTATGAGAA 7861 AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA 7921 ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC 7981 GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC 8041 CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT 8101 GCTCACATGT TCTTTCCTGC GTTATCCCCT GATTCTGTGG ATAACCGTAT TACCGCCTTT 8161 GAGTGAGCTG ATACCGCTCG CCGCAGCCGA ACGACCGAGC GCAGCGAGTC AGTGAGCGAG 8221 GAAGCGGAAG AGCGCCCAAT ACGCAAACCG CCTCTCCCCG CGCGTTGGCC GATTCATTAA 8281 TGCA //