LOCUS pLEW100cre 6643 bp DNA circular 20-JUL-2008 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 3146..3857 /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 4788..5645 /function="LACTAMASE" 3'UTR complement(744..1021) /function="ACT 3' UTR" CDS complement(1082..1456) /note="BLE" promoter complement(1599..1620) /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 1629..1877 /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." protein_bind 1883..1923 /function="Tet Operators" 5'UTR 1927..2012 /function="GPEET SAS" /note="To SAS" 5'UTR 2013..2043 /function="GPEET 5´ UTR" 5'UTR complement(1477..1593) /function="ACT 5' UTR & SAS" conflict 588..594 /note="The Sac site that the sequence indicates is not present" CDS 2103..3134 /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 2049..2102 /note="Extraneous vector" BASE COUNT 1679 a 1588 c 1722 g 1651 t 3 others 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 ATATAGCTTA TTTTATGGCA GCAACGAGAC CTTACGTAAA 1501 GCAAAACTAA ATAAGAGCGG AGACTGCAAT GCAGAGTAAA AAAAAAAAAA ACATGAATTT 1561 CAGAAGACCT TGCTGTGCCC CCGGTACGGG CTAGATCTCC CTATAGTGAG TCGTATTAAT 1621 CAGGTACCGT CATTGGGGTT AAGCGGAAAG GTGTGTGTCA GTAGGTTGTG AGGTGAAAGC 1681 GTTTTCAGAT GCATAGTGAG CTTAATGTCC TTTTCACAGT ATATCGTGTC TGATAGGTAT 1741 CTCTTATTAG TATAGTCGAA TACTAGTCAA TAGTGCGTTT TGTGCAAAAT GTCCATTTTG 1801 TGGCAGTGAT GGGGTTGTTT TATGCTATTC CGTGTCTCTG GGTGGGCGTG CATTGAAAAT 1861 AGGGGTTATC GGGTGAGGGA TCTCCCTATC AGTGATAGAG ATCTCCCTAT CAGTGATAGA 1921 GATCCCTGAG TACTGAGTTT AACATGTTCT CGTCCCGGGC TGCACGCGCC TTCGAGTTTT 1981 TTTTCCTTTT CCCCATTTTT TTCAACTTGA AGACTTCAAT TACACCAAAA AGTAAAATTC 2041 ACAAGCTCGA GGGGCAGAGC CGATCCTGTA CACTTTACTT AAAACCATTA TCTGAGTGTG 2101 AAATGTCCAA TTTACTGACC GTACACCAAA ATTTGCCTGC ATTACCGGTC GATGCaACGA 2161 GTGATGAGGT TCGCAAGAAC CTGATGGACA TGTTCAGGGA TCGCCAGGCG TTTTCTGAGC 2221 ATACCTGGAA AATGCTTCTG TCCGTTTGCC GGTCGTGGGC GGCATGGTGC AAGTTGAATA 2281 ACCGGAAATG GTTTCCCGCA GAACCTGAAG ATGTTCGCGA TTATCTTCTA TATCTTCAGG 2341 CGCGCGGTCT GGCAGTAAAA ACTATCCAGC AACATTTGGG CCAGCTAAAC ATGCTTCATC 2401 GTCGGTCCGG GCTGCCACGA CCAAGTGACA GCAATGCTGT TTCACTGGTT ATGCGGCGGA 2461 TCCGAAAAGA AAACGTTGAT GCCGGTGAAC GTGCAAAACA GGCTCTAGCG TTCGAACGCA 2521 CTGATTTCGA CCAGGTTCGT TCACTCATGG AAAATAGCGA TCGCTGCCAG GATATACGTA 2581 ATCTGGCATT TCTGGGGATT GCTTATAACA CCCTGTTACG TATAGCCGAA ATTGCCAGGA 2641 TCAGGGTTAA AGATATCTCA CGTACTGACG GTGGGAGAAT GTTAATCCAT ATTGGCAGAA 2701 CGAAAACGCT GGTTAGCACC GCAGGTGTAG AGAAGGCACT TAGCCTGGGG GTAACTAAAC 2761 TGGTCGAGCG ATGGATTTCC GTCTCTGGTG TAGCTGATGA TCCGAATAAC TACCTGTTTT 2821 GCCGGGTCAG AAAAAATGGT GTTGCCGCGC CATCTGCCAC CAGCCAGCTA TCAACTCGCG 2881 CCCTGGAAGG GATTTTTGAA GCAACTCATC GATTGATTTA CGGCGCTAAG GATGACTCTG 2941 GTCAGAGATA CCTGGCCTGG TCTGGACACA GTGCCCGTGT CGGAGCCGCG CGAGATATGG 3001 CCCGCGCTGG AGTTTCAATA CCGGAGATCA TGCAAGCTGG TGGCTGGACC AATGTAAATA 3061 TTGTCATGAA CTATATCCGT AACCTGGATA GTGAAACAGG GGCAATGGTG CGCCTGCTGG 3121 AAGATGGCGA TTAGCCATTA ACGCGGATCC TGCCCATTTA GTTGGCTTTT CCCTTGTCTC 3181 GTGTCTTTTC CGTGGAAAGG TTCCCGGAGT AATCTGATGG CACAGCAGGG AGGTGCGCCT 3241 GCAGGTTGGT TAGGAAGGGG GGATGATGTA AAAGAAGAAA ATGGGGGGAT ATCTTATGTT 3301 TAAGAGGATA AATAATGTGA AGGGGCTTTA TATGCTTGCT TGGTTGTCTC GTTGGTGCAT 3361 GGGGAATCTG CATGTTTGCT TTGGAGCACG CGTGGTACAT CAATGGGATC ATATCTTGCT 3421 GCCTCCCGCA GCTCACCGTG CGAGCTGCCG GGACCCCGTT TCTCATAGGC GTGTGCACCT 3481 TCCGTCTGAG ACCTGTAAAT GGTTAAAAAG GAATATATAA TTACCTTTTG AAAAGTGGTA 3541 AAAAACGAAT ATATCTTTTT TTTTGGTTTT AATACGCTCT TTTGTGTGTA TGAGAGGAAT 3601 AAATAAGTGT GTGTGTGTGT GTGTGTTGTT GTTGTTGAAA AAATGAAGGA CACGAAAGCG 3661 CTCATTAGGG GTGGGGATAC ACCAGGCATA ATTATCCGAA AATATTTTGA TGCACCAAAT 3721 AAGTGAACAT ACTGACGAAA TCAAAGAGTT GGAGGATGGA TAGGGAGGCC TCATTGGCAG 3781 TGGTAAATTG ATTTGACTTG AATATGATAC CCTCTTACTG TTTCATTGCC ACTACAAGTT 3841 GGTTTCCTTC CCCTGCAGGC ATGCAAGCTA GCTTGTATTC TATAGTGTCA CCTAAATCGT 3901 ATGTGTATGA TACATAAGGT TATGTATTAA TTGTAGCCGC GTTCTAACGA CAATATGTAC 3961 AAGCCTAATT GTGTAGCATC TGGCTTACTG AAGCAGACCC TATCATCTCT CTCGTAAACT 4021 GCCGTCAGAG TCGGTTTGGT TGGACGAACC TTCTGAGTTT CTGGTAACGC CGTTCCGCAC 4081 CCCGGAAATG GTCAGCGAAC CAATCAGCAG GGTCATCGCT AGCCAGATCC TCTACGCCGG 4141 ACGCATCGTG GCCGGCATCA CCGGCGCCAC AGGTGCGGTT GCTGGCGCCT ATATCGCCGA 4201 CATCACCGAT GGGGAAGATC GGGCTCGCCA CTTCGGGCTC ATGAGCGCTT GTTTCGGCGT 4261 GGGTATGGTG GCAGGCCCCG TGGCCGGGGG ACTGTTGGGC GCCATCTCCT TGCACCATTC 4321 CTTGCGGCGG CGGTGCTCAA CGGCCTCAAC CTACTACTGG GCTGCTTCCT AATGCAGGAG 4381 TCGCATAAGG GAGAGCGTCG ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT 4441 AGTTAAGCCA GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC 4501 TCCCGGCATC CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT 4561 TTTCACCGTC ATCACCGAAA CGCGCGAGAC GAAAGGGCCT CGTGATACGC CTATTTTTAT 4621 AGGTTAATGT CATGATAATA ATGGTTTCTT AGACGTCAGG TGGCACTTTT CGGGGAAATG 4681 TGCGCGGAAC CCCTATTTGT TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA 4741 GACAATAACC CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTATG AGTATTCAAC 4801 ATTTCCGTGT CGCCCTTATT CCCTTTTTTG CGGCATTTTG CCTTCCTGTT TTTGCTCACC 4861 CAGAAACGCT GGTGAAAGTA AAAGATGCTG AAGATCAGTT GGGTGCACGA GTGGGTTACA 4921 TCGAACTGGA TCTCAACAGC GGTAAGATCC TTGAGAGTTT TCGCCCCGAA GAACGTTTTC 4981 CAATGATGAG CACTTTTAAA GTTCTGCTAT GTGGCGCGGT ATTATCCCGT ATTGACGCCG 5041 GGCAAGAGCA ACTCGGTCGC CGCATACACT ATTCTCAGAA TGACTTGGTT GAGTACTCAC 5101 CAGTCACAGA AAAGCATCTT ACGGATGGCA TGACAGTAAG AGAATTATGC AGTGCTGCCA 5161 TAACCATGAG TGATAACACT GCGGCCAACT TACTTCTGAC AACGATCGGA GGACCGAAGG 5221 AGCTAACCGC TTTTTTGCAC AGCATGGGGG ATCATGTAAC TCGCCTTGAT CGTTGGGAAC 5281 CGGAGCTGAA TGAAGCCATA CCAAACGACG AGCGTGACAC CACGATGCCT GTAGCAATGG 5341 CAACAACGTT GCGCAAACTA TTAACTGGCG AACTACTTAC TCTAGCTTCC CGGCAACAAT 5401 TAATAGACTG GATGGAGGCG GATAAAGTTG CAGGACCACT TCTGCGCTCG GCCCTTCCGG 5461 CTGGCTGGTT TATTGCTGAT AAATCTGGAG CCGGTGAGCG TGGGTCTCGC GGTATCATTG 5521 CAGCACTGGG GCCAGATGGT AAGCCCTCCC GTATCGTAGT TATCTACACG ACGGGGAGTC 5581 AGGCAACTAT GGATGAACGA AATAGACAGA TCGCTGAGAT AGGTGCCTCA CTGATTAAGC 5641 ATTGGTAACT GTCAGACCAA GTTTACTCAT ATATACTTTA GATTGATTTA AAACTTCATT 5701 TTTAATTTAA AAGGATCTAG GTGAAGATCC TTTTTGATAA TCTCATGACC AAAATCCCTT 5761 AACGTGAGTT TTCGTTCCAC TGAGCGTCAG ACCCCGTAGA AAAGATCAAA GGATCTTCTT 5821 GAGATCCTTT TTTTCTGCGC GTAATCTGCT GCTTGCAAAC AAAAAAACCA CCGCTACCAG 5881 CGGTGGTTTG TTTGCCGGAT CAAGAGCTAC CAACTCTTTT TCCGAAGGTA ACTGGCTTCA 5941 GCAGAGCGCA GATACCAAAT ACTGTCCTTC TAGTGTAGCC GTAGTTAGGC CACCACTTCA 6001 AGAACTCTGT AGCACCGCCT ACATACCTCG CTCTGCTAAT CCTGTTACCA GTGGCTGCTG 6061 CCAGTGGCGA TAAGTCGTGT CTTACCGGGT TGGACTCAAG ACGATAGTTA CCGGATAAGG 6121 CGCAGCGGTC GGGCTGAACG GGGGGTTCGT GCACACAGCC CAGCTTGGAG CGAACGACCT 6181 ACACCGAACT GAGATACCTA CAGCGTGAGC TATGAGAAAG CGCCACGCTT CCCGAAGGGA 6241 GAAAGGCGGA CAGGTATCCG GTAAGCGGCA GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC 6301 TTCCAGGGGG AAACGCCTGG TATCTTTATA GTCCTGTCGG GTTTCGCCAC CTCTGACTTG 6361 AGCGTCGATT TTTGTGATGC TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC GCCAGCAACG 6421 CGGCCTTTTT ACGGTTCCTG GCCTTTTGCT GGCCTTTTGC TCACATGTTC TTTCCTGCGT 6481 TATCCCCTGA TTCTGTGGAT AACCGTATTA CCGCCTTTGA GTGAGCTGAT ACCGCTCGCC 6541 GCAGCCGAAC GACCGAGCGC AGCGAGTCAG TGAGCGAGGA AGCGGAAGAG CGCCCAATAC 6601 GCAAACCGCC TCTCCCCGCG CGTTGGCCGA TTCATTAATG CAG //