LOCUS pLEW100cre_del_tetO 6594 bp DNA circular 18-MAR-2010 FEATURES Location/Qualifiers misc_feature complement(8..712) /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." 3'UTR 3097..3808 /function="ALD 3' UTR" /note="long aldolase 3' UTR from pHD103 for optimized BF expression (about 6x higher than the short ALD UTR used in pLew79 & pLew82)." CDS 4739..5596 /function="LACTAMASE" 3'UTR complement(744..1021) /function="ACT 3' UTR" CDS complement(1082..1456) /note="BLE" promoter complement(1595..1616) /function="T7 Promoter" /note="T7 promoter, full-strength, constitutive. Note that there are no T7 terminators downstream of T7 transcription unit in this construct, which could be a problem in some insertion sites!" promoter 1625..1873 /function="GPEET Promoter" /note="Best match with GPEET2 promoter on chromosome 6: only 2 nt differ in 249 BLASTed. Originally Christine's PARP-A promoter region." 5'UTR 1879..1963 /function="GPEET SAS" /note="To SAS" 5'UTR 1964..1994 /function="GPEET 5%B4 UTR" 5'UTR complement(1475..1589) /function="ACT 5' UTR & SAS" conflict 588..594 /note="The Sac site that the sequence indicates is not present" CDS 2054..3085 /function="Cre Recombinase" /note="Cre was cloned into pLEW100 as a Hind-Bam fragment, but a one-nt change destroyed the HindIII site. Not clear how used Bam, as there is an internal site in Cre. On John's receommendation when he supplied the plasmid, we re-sequenced the gene. Ultimately, when making variants of it, we resequenced other bits, which explains why a few nt in this sequence differ from that given for the parental pLEW100 on this web site. Donelson's paper says Cre is the sequence from GenBank X03453, but they truncated that sequence in their vector. This ORF matches the ORF in X03453 and AF234173. The 5'UTR comes from whatever vector they used." misc_feature 2000..2053 /note="Extraneous vector" misc_feature 1444..3140 /note="verified sequence" 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 CGGCnnnCTT 721 GCATGCCTGC AAGGCCTTGC AGAATACTGC ATAGATAACA AACGCATCAA ACACAACTGG 781 TGGCACTATT TCAATCATGT CGACACACCA AGACGAGAAA ATTTGAGTTA AACAAGATGC 841 CTACAACCCT ATATTTATTA TCTTCAAGTG ATCTTCTCTT CTCTTGTGCG CTGTACGTAA 901 ATGTGTTGCA AATAAGGCAT AGGGCTGAGT ACAGGCACCA CACGCGAGGA AAATCATAAG 961 CACTAGCCGT GTCAACTCAC AGCGACTACA GAACAATTTT GGCCACACAA CCCGGTGTTA 1021 GGATCTCCGA GGCCTGGGAC CCGTGGGCCG CCGTCGGACC GGCGGTGTTG GTCGGCGTCG 1081 GTCAGTCCTG CTCCTCGGCC ACGAAGTGCA CGCAGTTGCC GGCCGGGTCG CGCAGGGCGA 1141 ACTCCCGCCC CCACGGCTGC TCGCCGATCT CGGTCATGGC CGGCCCGGAG GCGTCCCGGA 1201 AGTTCGTGGA CACGACCTCC GACCACTCGG CGTACAGCTC GTCCAGGCCG CGCACCCACA 1261 CCCAGGCCAG GGTGTTGTCC GGCACCACCT GGTCCTGGAC CGCGCTGATG AACAGGGTCA 1321 CGTCGTCCCG GACCACACCG GCGAAGTCGT CCTCCACGAA GTCCCGGGAG AACCCGAGCC 1381 GGTCGGTCCA GAACTCGACC GCTCCGGCGA CGTCGCGCGC GGTGAGCACC GGAACGGCAC 1441 TGGTCAACTT GGCCATGGTG ATAGCTTATT TTATGGCAGC AACGAGACCT TACGTAAAGC 1501 AAAACTAAAT AAGAGCGGAG ACTGCAATGC AGAGTAAAAA AAAAAAAAAC ATGAATTTCA 1561 GAAGACCTTG CTGTGCCCCC GGTACGGGAG ATCTCCCTAT AGTGAGTCGT ATTAATCAGG 1621 TACCGTCATT GGGGTTAAGC GGAAAGGTGT GTGTCAGTAG GTTGTGAGGT GAAAGCGTTT 1681 TCAGATGCAT AGTGAGCTTA ATGTCCTTTT CACAGTATAT CGTGTCTGAT AGGTATCTCT 1741 TATTAGTATA GTCGAATACT AGTCAATAGT GCGTTTTGTG CAAAATGTCC ATTTTGTGGC 1801 AGTGATGGGG TTGTTTTATG CTATTCCGTG TCTCTGGGTG GGCGTGCATT GAAAATAGGG 1861 GTTATCGGGT GAGGGATCGA GTACTGAGTT TAACATGTTC TCGTCCCGGG CTGCACGCGC 1921 CTTCGAGTTT TTTTTCCTTT TCCCCATTTT TTTCAACTTG AAGACTTCAA TTACACCAAA 1981 AAGTAAAATT CACAAGCTCG AGGGGCAGAG CCGATCCTGT ACACTTTACT TAAAACCATT 2041 ATCTGAGTGT GAAATGTCCA ATTTACTGAC CGTACACCAA AATTTGCCTG CATTACCGGT 2101 CGATGCAACG AGTGATGAGG TTCGCAAGAA CCTGATGGAC ATGTTCAGGG ATCGCCAGGC 2161 GTTTTCTGAG CATACCTGGA AAATGCTTCT GTCCGTTTGC CGGTCGTGGG CGGCATGGTG 2221 CAAGTTGAAT AACCGGAAAT GGTTTCCCGC AGAACCTGAA GATGTTCGCG ATTATCTTCT 2281 ATATCTTCAG GCGCGCGGTC TGGCAGTAAA AACTATCCAG CAACATTTGG GCCAGCTAAA 2341 CATGCTTCAT CGTCGGTCCG GGCTGCCACG ACCAAGTGAC AGCAATGCTG TTTCACTGGT 2401 TATGCGGCGG ATCCGAAAAG AAAACGTTGA TGCCGGTGAA CGTGCAAAAC AGGCTCTAGC 2461 GTTCGAACGC ACTGATTTCG ACCAGGTTCG TTCACTCATG GAAAATAGCG ATCGCTGCCA 2521 GGATATACGT AATCTGGCAT TTCTGGGGAT TGCTTATAAC ACCCTGTTAC GTATAGCCGA 2581 AATTGCCAGG ATCAGGGTTA AAGATATCTC ACGTACTGAC GGTGGGAGAA TGTTAATCCA 2641 TATTGGCAGA ACGAAAACGC TGGTTAGCAC CGCAGGTGTA GAGAAGGCAC TTAGCCTGGG 2701 GGTAACTAAA CTGGTCGAGC GATGGATTTC CGTCTCTGGT GTAGCTGATG ATCCGAATAA 2761 CTACCTGTTT TGCCGGGTCA GAAAAAATGG TGTTGCCGCG CCATCTGCCA CCAGCCAGCT 2821 ATCAACTCGC GCCCTGGAAG GGATTTTTGA AGCAACTCAT CGATTGATTT ACGGCGCTAA 2881 GGATGACTCT GGTCAGAGAT ACCTGGCCTG GTCTGGACAC AGTGCCCGTG TCGGAGCCGC 2941 GCGAGATATG GCCCGCGCTG GAGTTTCAAT ACCGGAGATC ATGCAAGCTG GTGGCTGGAC 3001 CAATGTAAAT ATTGTCATGA ACTATATCCG TAACCTGGAT AGTGAAACAG GGGCAATGGT 3061 GCGCCTGCTG GAAGATGGCG ATTAGCCATT AACGCGGATC CTGCCCATTT AGTTGGCTTT 3121 TCCCTTGTCT CGTGTCTTTT CCGTGGAAAG GTTCCCGGAG TAATCTGATG GCACAGCAGG 3181 GAGGTGCGCC TGCAGGTTGG TTAGGAAGGG GGGATGATGT AAAAGAAGAA AATGGGGGGA 3241 TATCTTATGT TTAAGAGGAT AAATAATGTG AAGGGGCTTT ATATGCTTGC TTGGTTGTCT 3301 CGTTGGTGCA TGGGGAATCT GCATGTTTGC TTTGGAGCAC GCGTGGTACA TCAATGGGAT 3361 CATATCTTGC TGCCTCCCGC AGCTCACCGT GCGAGCTGCC GGGACCCCGT TTCTCATAGG 3421 CGTGTGCACC TTCCGTCTGA GACCTGTAAA TGGTTAAAAA GGAATATATA ATTACCTTTT 3481 GAAAAGTGGT AAAAAACGAA TATATCTTTT TTTTTGGTTT TAATACGCTC TTTTGTGTGT 3541 ATGAGAGGAA TAAATAAGTG TGTGTGTGTG TGTGTGTTGT TGTTGTTGAA AAAATGAAGG 3601 ACACGAAAGC GCTCATTAGG GGTGGGGATA CACCAGGCAT AATTATCCGA AAATATTTTG 3661 ATGCACCAAA TAAGTGAACA TACTGACGAA ATCAAAGAGT TGGAGGATGG ATAGGGAGGC 3721 CTCATTGGCA GTGGTAAATT GATTTGACTT GAATATGATA CCCTCTTACT GTTTCATTGC 3781 CACTACAAGT TGGTTTCCTT CCCCTGCAGG CATGCAAGCT AGCTTGTATT CTATAGTGTC 3841 ACCTAAATCG TATGTGTATG ATACATAAGG TTATGTATTA ATTGTAGCCG CGTTCTAACG 3901 ACAATATGTA CAAGCCTAAT TGTGTAGCAT CTGGCTTACT GAAGCAGACC CTATCATCTC 3961 TCTCGTAAAC TGCCGTCAGA GTCGGTTTGG TTGGACGAAC CTTCTGAGTT TCTGGTAACG 4021 CCGTTCCGCA CCCCGGAAAT GGTCAGCGAA CCAATCAGCA GGGTCATCGC TAGCCAGATC 4081 CTCTACGCCG GACGCATCGT GGCCGGCATC ACCGGCGCCA CAGGTGCGGT TGCTGGCGCC 4141 TATATCGCCG ACATCACCGA TGGGGAAGAT CGGGCTCGCC ACTTCGGGCT CATGAGCGCT 4201 TGTTTCGGCG TGGGTATGGT GGCAGGCCCC GTGGCCGGGG GACTGTTGGG CGCCATCTCC 4261 TTGCACCATT CCTTGCGGCG GCGGTGCTCA ACGGCCTCAA CCTACTACTG GGCTGCTTCC 4321 TAATGCAGGA GTCGCATAAG GGAGAGCGTC GATATGGTGC ACTCTCAGTA CAATCTGCTC 4381 TGATGCCGCA TAGTTAAGCC AGCCCCGACA CCCGCCAACA CCCGCTGACG CGCCCTGACG 4441 GGCTTGTCTG CTCCCGGCAT CCGCTTACAG ACAAGCTGTG ACCGTCTCCG GGAGCTGCAT 4501 GTGTCAGAGG TTTTCACCGT CATCACCGAA ACGCGCGAGA CGAAAGGGCC TCGTGATACG 4561 CCTATTTTTA TAGGTTAATG TCATGATAAT AATGGTTTCT TAGACGTCAG GTGGCACTTT 4621 TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT CAAATATGTA 4681 TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA GGAAGAGTAT 4741 GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT GCCTTCCTGT 4801 TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT TGGGTGCACG 4861 AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT TTCGCCCCGA 4921 AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG TATTATCCCG 4981 TATTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA ATGACTTGGT 5041 TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA GAGAATTATG 5101 CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA CAACGATCGG 5161 AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAGCATGGGG GATCATGTAA CTCGCCTTGA 5221 TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA CCACGATGCC 5281 TGTAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA CTCTAGCTTC 5341 CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC TTCTGCGCTC 5401 GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC GTGGGTCTCG 5461 CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG TTATCTACAC 5521 GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA TAGGTGCCTC 5581 ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT AGATTGATTT 5641 AAAACTTCAT TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA ATCTCATGAC 5701 CAAAATCCCT TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG AAAAGATCAA 5761 AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA CAAAAAAACC 5821 ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT TTCCGAAGGT 5881 AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTCCTT CTAGTGTAGC CGTAGTTAGG 5941 CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA TCCTGTTACC 6001 AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA GACGATAGTT 6061 ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC CCAGCTTGGA 6121 GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CTATGAGAAA GCGCCACGCT 6181 TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG 6241 CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG GGTTTCGCCA 6301 CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC TATGGAAAAA 6361 CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG CTCACATGTT 6421 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA 6481 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA 6541 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAG //