LOCUS pLEW13 9261 bp DNA circular 16-MAR-2010 FEATURES Location/Qualifiers misc_feature 1..906 /function="%DF TUB targeting" CDS 4262..5128 /function="NEO" 3'UTR 5147..5435 /function="ACT 3' UTR" 5'UTR 5436..5555 /function="ACT SAS" 3'UTR 6197..6467 /function="ACT 3' UTR" 5'UTR 4094..4205 /function="ACT SAS" 3'UTR 3946..4090 /note="ALD 3' UTR" /note="truncated aldolase 3' UTR" 5'UTR 910..1106 /function="ALD SAS" /note="aldolase-derived sas from HD330" 5'UTR 1107..1198 /function="PARP SAS" /note="PARP-derived splice aceptor site from HD108" CDS 7406..8266 /note="LACTAMASE" misc_feature 1269..1305 /note="NLS" /note="36-nt insertion of SV40 T antigen NLS (JJ Dunn et al, 1988, Gene 68, 259-266)" CDS 1236..3923 /note="T7RNAP" /note="Sequence corrected October 2009 based on actual sequencing in ZM construct (formally 'pyrFEKO-HYG tetr-t7rnap actual correct'), which matches updated 2009 sequence in GenBank (FJ881694)." CDS 5572..6195 /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)" misc_feature 3944..3959 /note="" /note="Corrected after sequencing by Addgene March 2010. Deleted 3944-3982 & 3959-3966." ORIGIN 1 CTGACCGTAT CATGATGACT TTCTCCATCA TCCCATCCCC CAAGGTGTCC GACACTGTCG 61 TCGAGCCGTA CAATACGACT CTCTCCGTGC ACCAACTTGT GGAAAACTCC GATGAGTCGA 121 TGTGCATTGA CAACGAGGCA CTGTACGATA TTTGCTTCCG CACCCTGAAA CTGACAACAC 181 CAACGTTCGG TGACCTGAAC CACTTGGTGT CTGCTGTTGT GTCCGGCGTC ACCTGCTGCC 241 TGCGCTTCCC TGGTCAGTTG AACTCTGACC TCCGTAAGTT GGCTGTGAAC CTTGTCCCAT 301 TCCCGCGTCT GCACTTCTTC ATGATGGGCT TCGCCCCGCT GACCAGCCGC GGCTCGCAGC 361 AGTACCGCGG TCTCTCCGTG CCCGAGCTAA CGCAGCAGAT GTTCGATGCG AAAAACATGA 421 TGCAAGCTGC AGATCCTCGT CACGGCCGCT ACCTGACAGC GTCTGCACTC TTCCGCGGCC 481 GCATGTCGAC GAAGGAGGTT GATGAGCAGA TGCTGAACGT GCAGAACAAG AACTCGTCCT 541 ACTTCATTGA GTGGATCGAT CCCGAACAAC ATCAAGTCCT CTGTTTGCGA TATCCCACCC 601 AAGGGACTCA AGATGGCTGT CACCTTCATT GGCAACAACA CCTGCATCCA GGAGATGTTC 661 CGCCGTGTGG GAGAGCAGTT CACCCTCATG TTCCGTCGCA AGGCGTTCTT GCACTGGTAC 721 ACTGGCGAGG GTATGGACGA GATGGAATTC ACGGAGGCAG AGTCCAACAT GAACGATCTC 781 GTGTCTGAGT ACCAGCAGTA CCAGGATGCC ACGATTGAGG AGGAGGGCGA GTTCGACGAG 841 GAGGAGCAAT ACTAGACGCG GACGGGGCAT TTCCCGTTCG TCATTAGCAG TAGGTAATGA 901 AGATGCTCGA GGGTGCTCAA GCTGTGTAGC GCACGCGTTT CCTTACATAT TTCTCTAACA 961 GGCACGGAAG CCTAACAAAT ACACTTGGCT TATTTTTTTG CCCCCTCATG TCTTGTACAA 1021 ATATTTGCGA TAGCTTAGCT ATCAGCCACA TTAATCAAAC AAGTATACCA ACAAGCCCGA 1081 AAACATAAAC TCAACTGCAA CGAAGCTGGG CTGCACGCGC CTTCGAGTTT TTTTTCCTTT 1141 TCCCCATTTT TTTCAACTTG AAGACTTCAA TTACACCAAA AAGTAAAATT CACAAGCTGA 1201 TCTGATCCGG ATTTACTAAC TGGAAGAGGC ACTAAATGAA CACGATTAAC ATCGCTAAGA 1261 ACGAATTCCT CGAGCCTCCA AAAAAGAAGA GAAAGGTCGA ATTCTCTGAC ATCGAACTGG 1321 CTGCTATCCC GTTCAACACT CTGGCTGACC ATTACGGTGA GCGTTTAGCT CGCGAACAGT 1381 TGGCCCTTGA GCATGAGTCT TACGAGATGG GTGAAGCACG CTTCCGCAAG ATGTTTGAGC 1441 GTCAACTTAA AGCTGGTGAG GTTGCGGATA ACGCTGCCGC CAAGCCTCTC ATCACTACCC 1501 TACTCCCTAA GATGATTGCA CGCATCAACG ACTGGTTTGA GGAAGTGAAA GCTAAGCGCG 1561 GCAAGCGCCC GACAGCCTTC CAGTTCCTGC AAGAAATCAA GCCGGAAGCC GTAGCGTACA 1621 TCACCATTAA GACCACTCTG GCTTGCCTAA CCAGTGCTGA CAATACAACC GTTCAGGCTG 1681 TAGCAAGCGC AATCGGTCGG GCCATTGAGG ACGAGGCTCG CTTCGGTCGT ATCCGTGACC 1741 TTGAAGCTAA GCACTTCAAG AAAAACGTTG AGGAACAACT CAACAAGCGC GTAGGGCACG 1801 TCTACAAGAA AGCATTTATG CAAGTTGTCG AGGCTGACAT GCTCTCTAAG GGTCTACTCG 1861 GTGGCGAGGC GTGGTCTTCG TGGCATAAGG AAGACTCTAT TCATGTAGGA GTACGCTGCA 1921 TCGAGATGCT CATTGAGTCA ACCGGAATGG TTAGCTTACA CCGCCAAAAT GCTGGCGTAG 1981 TAGGTCAAGA CTCTGAGACT ATCGAACTCG CACCTGAATA CGCTGAGGCT ATCGCAACCC 2041 GTGCAGGTGC GCTGGCTGGC ATCTCTCCGA TGTTCCAACC TTGCGTAGTT CCTCCTAAGC 2101 CGTGGACTGG CATTACTGGT GGTGGCTATT GGGCTAACGG TCGTCGTCCT CTGGCGCTGG 2161 TGCGTACTCA CAGTAAGAAA GCACTGATGC GCTACGAAGA CGTTTACATG CCTGAGGTGT 2221 ACAAAGCGAT TAACATTGCG CAAAACACCG CATGGAAAAT CAACAAGAAA GTCCTAGCGG 2281 TCGCCAACGT AATCACCAAG TGGAAGCATT GTCCGGTCGA GGACATCCCT GCGATTGAGC 2341 GTGAAGAACT CCCGATGAAA CCGGAAGACA TCGACATGAA TCCTGAGGCT CTCACCGCGT 2401 GGAAACGTGC TGCCGCTGCT GTGTACCGCA AGGACAAGGC TCGCAAGTCT CGCCGTATCA 2461 GCCTTGAGTT CATGCTTGAG CAAGCCAATA AGTTTGCTAA CCATAAGGCC ATCTGGTTCC 2521 CTTACAACAT GGACTGGCGC GGTCGTGTTT ACGCTGTGTC AATGTTCAAC CCGCAAGGTA 2581 ACGATATGAC CAAAGGACTG CTTACGCTGG CGAAAGGTAA ACCAATCGGT AAGGAAGGTT 2641 ACTACTGGCT GAAAATCCAC GGTGCAAACT GTGCGGGTGT CGATAAGGTT CCGTTCCCTG 2701 AGCGCATCAA GTTCATTGAG GAAAACCACG AGAACATCAT GGCTTGCGCT AAGTCTCCAC 2761 TGGAGAACAC TTGGTGGGCT GAGCAAGATT CTCCGTTCTG CTTCCTTGCG TTCTGCTTTG 2821 AGTACGCTGG GGTACAGCAC CACGGCCTGA GCTATAACTG CTCCCTTCCG CTGGCGTTTG 2881 ACGGGTCTTG CTCTGGCATC CAGCACTTCT CCGCGATGCT CCGAGATGAG GTAGGTGGTC 2941 GCGCGGTTAA CTTGCTTCCT AGTGAAACCG TTCAGGACAT CTACGGGATT GTTGCTAAGA 3001 AAGTCAACGA GATTCTACAA GCAGACGCAA TCAATGGGAC CGATAACGAA GTAGTTACCG 3061 TGACCGATGA GAACACTGGT GAAATCTCTG AGAAAGTCAA GCTGGGCACT AAGGCACTGG 3121 CTGGTCAATG GCTGGCTTAC GGTGTTACTC GCAGTGTGAC TAAGCGTTCA GTCATGACGC 3181 TGGCTTACGG GTCCAAAGAG TTCGGCTTCC GTCAACAAGT GCTGGAAGAT ACCATTCAGC 3241 CAGCTATTGA TTCCGGCAAG GGTCTGATGT TCACTCAGCC GAATCAGGCT GCTGGATACA 3301 TGGCTAAGCT GATTTGGGAA TCTGTGAGCG TGACGGTGGT AGCTGCGGTT GAAGCAATGA 3361 ACTGGCTTAA GTCTGCTGCT AAGCTGCTGG CTGCTGAGGT CAAAGATAAG AAGACTGGAG 3421 AGATTCTTCG CAAGCGTTGC GCTGTGCATT GGGTAACTCC TGATGGTTTC CCTGTGTGGC 3481 AGGAATACAA GAAGCCTATT CAGACGCGCT TGAACCTGAT GTTCCTCGGT CAGTTCCGCT 3541 TACAGCCTAC CATTAACACC AACAAAGATA GCGAGATTGA TGCACACAAA CAGGAGTCTG 3601 GTATCGCTCC TAACTTTGTA CACAGCCAAG ACGGTAGCCA CCTTCGTAAG ACTGTAGTGT 3661 GGGCACACGA GAAGTACGGA ATCGAATCTT TTGCACTGAT TCACGACTCC TTCGGTACCA 3721 TTCCGGCTGA CGCTGCGAAC CTGTTCAAAG CAGTGCGCGA AACTATGGTT GACACATATG 3781 AGTCTTGTGA TGTACTGGCT GATTTCTACG ACCAGTTCGC TGACCAGTTG CACGAGTCTC 3841 AATTGGACAA AATGCCAGCA CTTCCGGCTA AAGGTAACTT GAACCTCCGT GACATCTTAG 3901 AGTCGGACTT CGCGTTCGCG TAACGCCAAA TCAATACGAC TCCGGATCCT GCCCATTTGG 3961 CTTTTCCCTT GTCTCGTGTC TTTTCCGTGG AAAGGTTCCC GGAGTAATCT GATGGCACAG 4021 CAGGGAGGTG CGCCTGCAGG TTGGTTAGGA AGGGGGGATG ATGTAAAAGA AGAAAATGGG 4081 GGGATTCGAG cccgggcaca gcaaggtctt ctgaaattca tgtttttttt ttttttactc 4141 tgcattgcag tctccgctct tatttagttt tgctttacgt aaggtctcgt tgctgccata 4201 aaataagctt agaggagaaa gaatagtaac cctttcatca aagaaaatag ttcaaacgaa 4261 tatgcgcgaa atcgtctgcg ttcaggctgg ccaatgcggt aaccagatcg gctcaaagtt 4321 ctgggaggtg atccggccaa gcttggatgg attgcacgca ggttctccgg ccgcttgggt 4381 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 4441 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 4501 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 4561 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 4621 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 4681 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 4741 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 4801 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 4861 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 4921 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 4981 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 5041 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 5101 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcggatc ctaacaccgg 5161 gttgtgtggc caaaattgtt ctgtagtcgc tgtgagttga cacggctagt gcttatgatt 5221 ttcctcgcgt gtggtgcctg tactcagccc tatgccttat ttgcaacaca tttacgtaca 5281 gcgcacaaga gaagagaaga tcacttgaag ataataaata tagggttgta ggcatcttgt 5341 ttaactcaaa ttttctcgtc ttggtgtgtc gacatgattg aaatagtgcc accagttgtg 5401 tttgatgcgt ttgttatcta tgcagtattc tgcaaggggg cacagcaagg tcttctgaaa 5461 ttcatgtttt tttttttttt actctgcatt gcagtctccg ctcttattta gttttgcttt 5521 acgtaaggtc tcgttgctgc cataaaataa gcctaGAACT AGAGCTTTCA TATGTCTAGA 5581 TTAGATAAAA GTAAAGTGAT TAACAGCGCA TTAGAGCTGC TTAATGAGGT CGGAATCGAA 5641 GGTTTAACAA CCCGTAAACT CGCCCAGAAG CTAGGTGTAG AGCAGCCTAC ATTGTATTGG 5701 CATGTAAAAA ATAAGCGGGC TTTGCTCGAC GCCTTAGCCA TTGAGATGTT AGATAGGCAC 5761 CATACTCACT TTTGCCCTTT AGAAGGGGAA AGCTGGCAAG ATTTTTTACG TAATAACGCT 5821 AAAAGTTTTA GATGTGCTTT ACTAAGTCAT CGCGATGGAG CAAAAGTACA TTTAGGTACA 5881 CGGCCTACAG AAAAACAGTA TGAAACTCTC GAAAATCAAT TAGCCTTTTT ATGCCAACAA 5941 GGTTTTTCAC TAGAGAATGC ATTATATGCA CTCAGCGCTG TGGGGCATTT TACTTTAGGT 6001 TGCGTATTGG AAGATCAAGA GCATCAAGTC GCTAAAGAAG AAAGGGAAAC ACCTACTACT 6061 GATAGTATGC CGCCATTATT ACGACAAGCT ATCGAATTAT TTGATCACCA AGGTGCAGAG 6121 CCAGCCTTCT TATTCGGCCT TGAATTGATC ATATGCGGAT TAGAAAAACA ACTTAAATGT 6181 GAAAGTGGGT CTTAAGGATC CCAAAATTGT TCTGTAGTCG CTGTGAGTTG ACACGGCTAG 6241 TGCTTATGAT TTTCCTCGCG TGTGGTGCCT GTACTCAGCC CTATGCCTTA TTTGCAACAC 6301 ATTTACGTAC AGCGCACAAG AGAAGAGAAG ATCACTTGAA GATAATAAAT ATAGGGTTGT 6361 AGGCATCTTG TTTAACTCAA ATTTTCTCGT CTTGGTGTGT CGACATGATT GAAATAGTGC 6421 CACCAGTTGT GTTTGATGCG TTTGTTATCT ATGCAGTATT CTGCAAGGCC TTGCAGGCAT 6481 GCAAGCTAGC TTGTATTCTA TAGTGTCACC TAAATCGTAT GTGTATGATA CATAAGGTTA 6541 TGTATTAATT GTAGCCGCGT TCTAACGACA ATATGTACAA GCCTAATTGT GTAGCATCTG 6601 GCTTACTGAA GCAGACCCTA TCATCTCTCT CGTAAACTGC CGTCAGAGTC GGTTTGGTTG 6661 GACGAACCTT CTGAGTTTCT GGTAACGCCG TTCCGCACCC CGGAAATGGT CAGCGAACCA 6721 ATCAGCAGGG TCATCGCTAG CCAGATCCTC TACGCCGGAC GCATCGTGGC CGGCATCACC 6781 GGCGCCACAG GTGCGGTTGC TGGCGCCTAT ATCGCCGACA TCACCGATGG GGAAGATCGG 6841 GCTCGCCACT TCGGGCTCAT GAGCGCTTGT TTCGGCGTGG GTATGGTGGC AGGCCCCGTG 6901 GCCGGGGGAC TGTTGGGCGC CATCTCCTTG CACCATTCCT TGCGGCGGCG GTGCTCAACG 6961 GCCTCAACCT ACTACTGGGC TGCTTCCTAA TGCAGGAGTC GCATAAGGGA GAGCGTCGAT 7021 ATGGTGCACT CTCAGTACAA TCTGCTCTGA TGCCGCATAG TTAAGCCAGC CCCGACACCC 7081 GCCAACACCC GCTGACGCGC CCTGACGGGC TTGTCTGCTC CCGGCATCCG CTTACAGACA 7141 AGCTGTGACC GTCTCCGGGA GCTGCATGTG TCAGAGGTTT TCACCGTCAT CACCGAAACG 7201 CGCGAGACGA AAGGGCCTCG TGATACGCCT ATTTTTATAG GTTAATGTCA TGATAATAAT 7261 GGTTTCTTAG ACGTCAGGTG GCACTTTTCG GGGAAATGTG CGCGGAACCC CTATTTGTTT 7321 ATTTTTCTAA ATACATTCAA ATATGTATCC GCTCATGAGA CAATAACCCT GATAAATGCT 7381 TCAATAATAT TGAAAAAGGA AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC 7441 CTTTTTTGCG GCATTTTGCC TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA 7501 AGATGCTGAA GATCAGTTGG GTGCACGAGT GGGTTACATC GAACTGGATC TCAACAGCGG 7561 TAAGATCCTT GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA ATGATGAGCA CTTTTAAAGT 7621 TCTGCTATGT GGCGCGGTAT TATCCCGTAT TGACGCCGGG CAAGAGCAAC TCGGTCGCCG 7681 CATACACTAT TCTCAGAATG ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC 7741 GGATGGCATG ACAGTAAGAG AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC 7801 GGCCAACTTA CTTCTGACAA CGATCGGAGG ACCGAAGGAG CTAACCGCTT TTTTGCACAA 7861 CATGGGGGAT CATGTAACTC GCCTTGATCG TTGGGAACCG GAGCTGAATG AAGCCATACC 7921 AAACGACGAG CGTGACACCA CGATGCCTGT AGCAATGGCA ACAACGTTGC GCAAACTATT 7981 AACTGGCGAA CTACTTACTC TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA 8041 TAAAGTTGCA GGACCACTTC TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA 8101 ATCTGGAGCC GGTGAGCGTG GGTCTCGCGG TATCATTGCA GCACTGGGGC CAGATGGTAA 8161 GCCCTCCCGT ATCGTAGTTA TCTACACGAC GGGGAGTCAG GCAACTATGG ATGAACGAAA 8221 TAGACAGATC GCTGAGATAG GTGCCTCACT GATTAAGCAT TGGTAACTGT CAGACCAAGT 8281 TTACTCATAT ATACTTTAGA TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT 8341 GAAGATCCTT TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG 8401 AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT 8461 AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA 8521 AGAGCTACCA ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC 8581 TGTCCTTCTA GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC 8641 ATACCTCGCT CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT 8701 TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG 8761 GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA 8821 GCGTGAGCTA TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT 8881 AAGCGGCAGG GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA 8941 TCTTTATAGT CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC 9001 GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC 9061 CTTTTGCTGG CCTTTTGCTC ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA 9121 CCGTATTACC GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG 9181 CGAGTCAGTG AGCGAGGAAG CGGAAGAGCG CCCAATACGC AAACCGCCTC TCCCCGCGCG 9241 TTGGCCGATT CATTAATGCA G //