LOCUS pLEW114-HYG5' 5236 bp DNA circular 22-OCT-2009 Used to make 'single marker' cell lines. Cells alrady transfected with pHD328 were transfected with this plasmid linearized with AatII. See Wirtz et al 1999. FEATURES Location/Qualifiers CDS 9..261 /note="HYG 5'" /note="H3-EcoRI fragment from hyg gene in p72hyg72 (pos. 2 to 250 of hyg) was cloned blunt into the pLew114 cut with XhoI and AflIII (filled in with Klenow), replacing the tubulin targeting segment." promoter 297..316 /function="10% T7 promoter" /note="10% mutant T7 promoter and initiation region. Cloned in filled Mlu of ald sas as annealed oligo cassette with filled Age overhangs" 5'UTR 267..502 /function="ALD SAS and 5' UTR" CDS 1452..2318 /function="NEO" 3'UTR 2319..2627 /function="ACT 3' UTR" 5'UTR 1284..1451 /function="ACT SAS and 5' UTR" 3'UTR 1136..1280 /function="ALD 3' UTR" CDS 507..1130 /function="Tet repressor" /note="CDS corrected 091022 to match the actually sequenced (November 2007) version in pLEW29, which is also a perfect match for sequences in GenBank (checked 091022)." BASE COUNT 1271 a 1254 c 1360 g 1351 t ORIGIN 1 CTCACATGAG CTTATGAAAA AGCCTGAACT CACCGCGACG TCTGTCGAGA AGTTTCTGAT 61 CGAAAAGTTC GACAGCGTCT CCGACCTGAT GCAGCTCTCG GAGGGCGAAG AATCTCGTGC 121 TTTCAGCTTC GATGTAGGAG GGCGTGGATA TGTCCTGCGG GTAAATAGCT GCGCCGATGG 181 TTTCTACAAA GATCGTTATG TTTATCGGCA CTTTGCATCG GCCGCGCTCC CGATTCCGGA 241 AGTGCTTGAC ATTGGGGAAT TTCGAGGGTG CTCAAGCTGT GTAGCGCACG CGCCGGTAAT 301 ACGTCTCACT ATAGGGAGAT CTACCGGCGC GTTTCCTTAC ATATTTCTCT AACAGGCACG 361 GAAGCCTAAC AAATACACTT GGCTTATTTT TTTGCCCCCT CATGTCTTGT ACAAATATTT 421 GCGATAGCTT AGCTATCAGC CACATTAATC AAACAAGTAT ACCAACAAGC CCGAAAACAT 481 AAACTCAACT GCAACGAAGC TTTCATATGT CTAGATTAGA TAAAAGTAAA GTGATTAACA 541 GCGCATTAGA GCTGCTTAAT GAGGTCGGAA TCGAAGGTTT AACAACCCGT AAACTCGCCC 601 AGAAGCTAGG TGTAGAGCAG CCTACATTGT ATTGGCATGT AAAAAATAAG CGGGCTTTGC 661 TCGACGCCTT AGCCATTGAG ATGTTAGATA GGCACCATAC TCACTTTTGC CCTTTAGAAG 721 GGGAAAGCTG GCAAGATTTT TTACGTAATA ACGCTAAAAG TTTTAGATGT GCTTTACTAA 781 GTCATCGCGA TGGAGCAAAA GTACATTTAG GTACACGGCC TACAGAAAAA CAGTATGAAA 841 CTCTCGAAAA TCAATTAGCC TTTTTATGCC AACAAGGTTT TTCACTAGAG AATGCATTAT 901 ATGCACTCAG CGCTGTGGGG CATTTTACTT TAGGTTGCGT ATTGGAAGAT CAAGAGCATC 961 AAGTCGCTAA AGAAGAAAGG GAAACACCTA CTACTGATAG TATGCCGCCA TTATTACGAC 1021 AAGCTATCGA ATTATTTGAT CACCAAGGTG CAGAGCCAGC CTTCTTATTC GGCCTTGAAT 1081 TGATCATATG CGGATTAGAA AAACAACTTA AATGTGAAAG TGGGTCTTAA GATCCTGCCC 1141 ATTTAGTTGG CTTTTCCCTT GTCTCGTGTC TTTTCCGTGG AAAGGTTCCC GGAGTAATCT 1201 GATGGCACAG CAGGGAGGTG CGCCTGCAGG TTGGTTAGGA AGGGGGGATG ATGTAAAAGA 1261 AGAAAATGGG GGGATTCGAG CCCGGGCACA GCAAGGTCTT CTGAAATTCA TGTTTTTTTT 1321 TTTTTTACTC TGCATTGCAG TCTCCGCTCT TATTTAGTTT TGCTTTACGT AAGGTCTCGT 1381 TGCTGCCATA AAATAAGCTT AGAGGAGAAA GAATAGTAAC CCTTTCATCA AAGAAAATAG 1441 TTCAAACGAA TATGCGCGAA ATCGTCTGCG TTCAGGCTGG CCAATGCGGT AACCAGATCG 1501 GCTCAAAGTT CTGGGAGGTG ATCCGGCCAA GCTTGGATGG ATTGCACGCA GGTTCTCCGG 1561 CCGCTTGGGT GGAGAGGCTA TTCGGCTATG ACTGGGCACA ACAGACAATC GGCTGCTCTG 1621 ATGCCGCCGT GTTCCGGCTG TCAGCGCAGG GGCGCCCGGT TCTTTTTGTC AAGACCGACC 1681 TGTCCGGTGC CCTGAATGAA CTGCAGGACG AGGCAGCGCG GCTATCGTGG CTGGCCACGA 1741 CGGGCGTTCC TTGCGCAGCT GTGCTCGACG TTGTCACTGA AGCGGGAAGG GACTGGCTGC 1801 TATTGGGCGA AGTGCCGGGG CAGGATCTCC TGTCATCTCA CCTTGCTCCT GCCGAGAAAG 1861 TATCCATCAT GGCTGATGCA ATGCGGCGGC TGCATACGCT TGATCCGGCT ACCTGCCCAT 1921 TCGACCACCA AGCGAAACAT CGCATCGAGC GAGCACGTAC TCGGATGGAA GCCGGTCTTG 1981 TCGATCAGGA TGATCTGGAC GAAGAGCATC AGGGGCTCGC GCCAGCCGAA CTGTTCGCCA 2041 GGCTCAAGGC GCGCATGCCC GACGGCGAGG ATCTCGTCGT GACCCATGGC GATGCCTGCT 2101 TGCCGAATAT CATGGTGGAA AATGGCCGCT TTTCTGGATT CATCGACTGT GGCCGGCTGG 2161 GTGTGGCGGA CCGCTATCAG GACATAGCGT TGGCTACCCG TGATATTGCT GAAGAGCTTG 2221 GCGGCGAATG GGCTGACCGC TTCCTCGTGC TTTACGGTAT CGCCGCTCCC GATTCGCAGC 2281 GCATCGCCTT CTATCGCCTT CTTGACGAGT TCTTCTGAGC GGGACTCTGG GGTTCGGATC 2341 CTAACACCGG GTTGTGTGGC CAAAATTGTT CTGTAGTCGC TGTGAGTTGA CACGGCTAGT 2401 GCTTATGATT TTCCTCGCGT GTGGTGCCTG TACTCAGCCC TATGCCTTAT TTGCAACACA 2461 TTTACGTACA GCGCACAAGA GAAGAGAAGA TCACTTGAAG ATAATAAATA TAGGGTTGTA 2521 GGCATCTTGT TTAACTCAAA TTTTCTCGTC TTGGTGTGTC GACATGATTG AAATAGTGCC 2581 ACCAGTTGTG TTTGATGCGT TTGTTATCTA TGCAGTATTC TGCAAGGCCT TGCAGGCATG 2641 CAAGCTAGCT TGTATTCTAT AGTGTCACCT AAATCGTATG TGTATGATAC ATAAGGTTAT 2701 GTATTAATTG TAGCCGCGTT CTAACGACAA TATGTACAAG CCTAATTGTG TAGCATCTGG 2761 CTTACTGAAG CAGACCCTAT CATCTCTCTC GTAAACTGCC GTCAGAGTCG GTTTGGTTGG 2821 ACGAACCTTC TGAGTTTCTG GTAACGCCGT TCCGCACCCC GGAAATGGTC AGCGAACCAA 2881 TCAGCAGGGT CATCGCTAGC CAGATCCTCT ACGCCGGACG CATCGTGGCC GGCATCACCG 2941 GCGCCACAGG TGCGGTTGCT GGCGCCTATA TCGCCGACAT CACCGATGGG GAAGATCGGG 3001 CTCGCCACTT CGGGCTCATG AGCGCTTGTT TCGGCGTGGG TATGGTGGCA GGCCCCGTGG 3061 CCGGGGGACT GTTGGGCGCC ATCTCCTTGC ACCATTCCTT GCGGCGGCGG TGCTCAACGG 3121 CCTCAACCTA CTACTGGGCT GCTTCCTAAT GCAGGAGTCG CATAAGGGAG AGCGTCGATA 3181 TGGTGCACTC TCAGTACAAT CTGCTCTGAT GCCGCATAGT TAAGCCAGCC CCGACACCCG 3241 CCAACACCCG CTGACGCGCC CTGACGGGCT TGTCTGCTCC CGGCATCCGC TTACAGACAA 3301 GCTGTGACCG TCTCCGGGAG CTGCATGTGT CAGAGGTTTT CACCGTCATC ACCGAAACGC 3361 GCGAGACGAA AGGGCCTCGT GATACGCCTA TTTTTATAGG TTAATGTCAT GATAATAATG 3421 GTTTCTTAGA CGTCAGGTGG CACTTTTCGG GGAAATGTGC GCGGAACCCC TATTTGTTTA 3481 TTTTTCTAAA TACATTCAAA TATGTATCCG CTCATGAGAC AATAACCCTG ATAAATGCTT 3541 CAATAATATT GAAAAAGGAA GAGTATGAGT ATTCAACATT TCCGTGTCGC CCTTATTCCC 3601 TTTTTTGCGG CATTTTGCCT TCCTGTTTTT GCTCACCCAG AAACGCTGGT GAAAGTAAAA 3661 GATGCTGAAG ATCAGTTGGG TGCACGAGTG GGTTACATCG AACTGGATCT CAACAGCGGT 3721 AAGATCCTTG AGAGTTTTCG CCCCGAAGAA CGTTTTCCAA TGATGAGCAC TTTTAAAGTT 3781 CTGCTATGTG GCGCGGTATT ATCCCGTATT GACGCCGGGC AAGAGCAACT CGGTCGCCGC 3841 ATACACTATT CTCAGAATGA CTTGGTTGAG TACTCACCAG TCACAGAAAA GCATCTTACG 3901 GATGGCATGA CAGTAAGAGA ATTATGCAGT GCTGCCATAA CCATGAGTGA TAACACTGCG 3961 GCCAACTTAC TTCTGACAAC GATCGGAGGA CCGAAGGAGC TAACCGCTTT TTTGCACAAC 4021 ATGGGGGATC ATGTAACTCG CCTTGATCGT TGGGAACCGG AGCTGAATGA AGCCATACCA 4081 AACGACGAGC GTGACACCAC GATGCCTGTA GCAATGGCAA CAACGTTGCG CAAACTATTA 4141 ACTGGCGAAC TACTTACTCT AGCTTCCCGG CAACAATTAA TAGACTGGAT GGAGGCGGAT 4201 AAAGTTGCAG GACCACTTCT GCGCTCGGCC CTTCCGGCTG GCTGGTTTAT TGCTGATAAA 4261 TCTGGAGCCG GTGAGCGTGG GTCTCGCGGT ATCATTGCAG CACTGGGGCC AGATGGTAAG 4321 CCCTCCCGTA TCGTAGTTAT CTACACGACG GGGAGTCAGG CAACTATGGA TGAACGAAAT 4381 AGACAGATCG CTGAGATAGG TGCCTCACTG ATTAAGCATT GGTAACTGTC AGACCAAGTT 4441 TACTCATATA TACTTTAGAT TGATTTAAAA CTTCATTTTT AATTTAAAAG GATCTAGGTG 4501 AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC GTTCCACTGA 4561 GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA 4621 ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA 4681 GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT ACCAAATACT 4741 GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC ACCGCCTACA 4801 TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA GTCGTGTCTT 4861 ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG 4921 GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG 4981 CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG GTATCCGGTA 5041 AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT 5101 CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT GTGATGCTCG 5161 TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC 5221 TTTTGCTGGC CTTTTG //