LOCUS pyrFEKO-BSD 5553 bp DNA circular 28-JUL-2008 The two PvuII sites that flank the PYR-FE targeting sequences are not shown on the map. FEATURES Location/Qualifiers misc_feature 2197..2693 /function="PYR-FE Downstream" /note="Sequence updated to agree with results of sequence done by Mike 21 Dec 06. Some minor disagreements with Sanger TbDB sequence. " CDS 3698..4555 /function="LACTAMASE" misc_feature 4..369 /note="PYR-FE upstream" /note="ends 136 bp before start of CDS." protein_bind 376..409 /function="loxP" 5'UTR 437..522 /function="Procyclin SAS and UTR" /note="The various procyclin SASs differ by onl 1 nt, really. This sequence taken from pNS10-54, with hindIII site removed (AAGCTT > AAGACT, leaving splice acceptor AG intact)" 5'UTR 523..580 /function="Residual UTR" /note="With minor changes to delete the infamous 'upstream ATG' CDS and about 4 deletions/insertions/changes to remove or create restriction sites, compared to the version in pNS10-54, from which Mike got the sequence." 3'UTR 2702..2760 /note="ALD 3' UTR (part)" /note="starts 96 bp after aldolase stop codon. Note that the SbfI (CCTGCAGG) site occurs in the aldolase 3' utr sequence at this point, so will be in lots of our plasmids." CDS 582..977 /note="BSD" CDS 984..1013 /function="Ty-1 epitope" CDS 582..2150 /function="BSD-TK" CDS 1020..2150 /note="HSVTK" protein_bind 2157..2190 /function="loxP" BASE COUNT 1278 a 1445 c 1452 g 1378 t ORIGIN 1 CTGTGGCGAA TAATTGTAGC GGCAGTTTCT AACGGTTATA TGTTTAACTG CATTATGCCC 61 CTCTCTCACT TGGCGCCATC TCTGTGTTAC AACATGCGTT TGATCTATCA GTTTATTATA 121 TTGTTTTCCC TCAAACACAT GCGAAGCGAG TAAGAGTTGA AAATGTTACG GCAAAGGCCA 181 ATGGGCACTG GTAGAAGGCA ACGGATCGCG GTTATGAAAG TTCTTGTGTG TATGTTTATG 241 TGGGGGTGGG GGTGGAAGTT GTGGTAGTGT CATCTCGTGT ACATTGTTTA AGTCGCTTCA 301 ACCTTGTTTC TTTTTTTTTT CCTTTTCTTC TTCTTCGCTA TCTGATTGAG TTTTCATACT 361 AAGTTGGATA AGCTTATAAC TTCGTATAGC ATACATTATA CGAAGTTATa ccggtACAcG 421 TTCTCGTCgg ccggccGCTG CACGCGCCTT CGAGTTTTTT TTCCTTTTCC CCATTTTTTT 481 CAACTTGAAG aCTTCAATTA CACCAAAAAA TAAAATTCAC AAACTTGGtA TTCCTTTGTG 541 TTACATTCTT GATCGCTCGC ACTGACATTA CagatctCAG CATGGCCAAG CCTTTGTCTC 601 AAGAAGAATC CACCCTCATT GAAAGAGCAA CGGCTACAAT CAACAGCATC CCCATCTCTG 661 AAGACTACAG CGTCGCCAGC GCAGCTCTCT CTAGCGACGG CCGCATCTTC ACTGGTGTCA 721 ATGTATATCA TTTTACTGGG GGACCTTGTG CAGAACTCGT GGTGCTGGGC ACTGCTGCTG 781 CTGCGGCGGC TGGCAACCTG ACTTGTATCG TCGCGATCGG AAATGAGAAC AGGGGCATCT 841 TGAGCCCCTG CGGACGGTGC CGACAGGTGC TTCTCGATCT GCATCCTGGG ATCAAAGCCA 901 TAGTGAAGGA CAGTGATGGA CAGCCGACGG CAGTTGGGAT TCGTGAATTG CTGCCCTCTG 961 GTTATGTGTG GGAGGGCTCT AGAGAAGTCC ATACTAACCA GGACCCACTT GACGCCACCA 1021 TGGCCTCGTA CCCCGGCCAT CAACACGCGT CTGCGTTCGA CCAGGCTGCG CGTTCTCGCG 1081 GCCATAGCAA CCGACGTACG GCGTTGCGCC CTCGCCGGCA GCAAGAAGCC ACGGAAGTCC 1141 GCCCGGAGCA GAAAATGCCC ACGCTACTGC GGGTTTATAT AGACGGTCCC CACGGGATGG 1201 GGAAAACCAC CACCACGCAA CTGCTGGTGG CCCTGGGTTC GCGCGACGAT ATCGTCTACG 1261 TACCCGAGCC GATGACTTAC TGGCGGGTGC TGGGGGCTTC CGAGACAATC GCGAACATCT 1321 ACACCACACA ACACCGCCTC GACCAGGGTG AGATATCGGC CGGGGACGCG GCGGTGGTAA 1381 TGACAAGCGC CCAGATAACA ATGGGCATGC CTTATGCCGT GACCGACGCC GTTCTGGCTC 1441 CTCATATCGG GGGGGAGGCT GGGAGCTCAC ATGCCCCGCC CCCGGCCCTC ACCCTCATCT 1501 TCGACCGCCA TCCCATCGCC GCCCTCCTGT GCTACCCGGC CGCGCGGTAC CTTATGGGCA 1561 GCATGACCCC CCAGGCCGTG CTGGCGTTCG TGGCCCTCAT CCCGCCGACC TTGCCCGGCA 1621 CCAACATCGT GCTTGGGGCC CTTCCGGAGG ACAGACACAT CGACCGCCTG GCCAAACGCC 1681 AGCGCCCCGG CGAGCGGCTG GACCTGGCTA TGCTGGCTGC GATTCGCCGC GTTTACGGGC 1741 TACTTGCCAA TACGGTGCGG TATCTGCAGT GCGGCGGGTC GTGGCGGGAG GACTGGGGAC 1801 AGCTTTCGGG GACGGCCGTG CCGCCCCAGG GTGCCGAGCC CCAGAGCAAC GCGGGCCCAC 1861 GACCCCATAT CGGGGACACG TTATTTACCC TGTTTCGGGC CCCCGAGTTG CTGGCCCCCA 1921 ACGGCGACCT GTATAACGTG TTTGCCTGGG CCTTGGACGT CTTGGCCAAA CGCCTCCGTT 1981 CCATGCACGT CTTTATCCTG GATTACGACC AATCGCCCGC CGGCTGCCGG GACGCCCTGC 2041 TGCAACTTAC CTCCGGGATG GTCCAGACCC ACGTCACCAC CCCCGGCTCC ATACCGACGA 2101 TATGCGACCT GGCGCGCACG TTTGCCCGGG AGATGGGGGA GGCTAACTGA GAATTCATAA 2161 CTTCGTATAG CATACATTAT ACGAAGTTAT GGATCCTAAG TGGGTCGGAG CTCTATCTTT 2221 AAGCTATTGC GGACGTACAC ACATGTTTTC GTTGCAAATT ATTTTCGtAC CTACTTCAGA 2281 TCGTAAGCGT GGGGAATAAT AATAATGCTC CCTACCATGA ATTTAAaCAG TTTGGTGAAT 2341 GAACAATTCa CTTTAAATTG ATGGACCATG AAATGCACTT TTACGGTCGC AGCGTTTAAC 2401 TAAGTGGCGA aGGCAAGTTT TTAATAATAA TAATAAAGCA AgTAGTAAAC TATATGAGAC 2461 AGCAATGGGG TTTGGGAGGG AAGGTTTAAT CGCTTCAAAG GTATTTGTGT GTGGTTGAGG 2521 AGGTGATAGC GAAGTGAGGG TTTCTAATAA CTGTAGAGCA GCAATAAAAA AAAGGTAGCA 2581 GTTGATCAAT TTGCTGTGGT GCCCTCTGCA TTGAAGGGTA TCTGGGATGT ATGAAGTTCC 2641 ATCACTCGGA GCCATCACCT TTCCCTCATT TCTCGTTTAC ATCCTTTACA TGTCAGCTGC 2701 CTGCAGGTTG GTTAGGAAGG GGGGATGATG TAAAAGAAGA AAATGGGGGG ATTCGAGCCC 2761 CCTTGCAGGC ATGCAAGCTA GCTTGTATTC TATAGTGTCA CCTAAATCGT ATGTGTATGA 2821 TACATAAGGT TATGTATTAA TTGTAGCCGC GTTCTAACGA CAATATGTAC AAGCCTAATT 2881 GTGTAGCATC TGGCTTACTG AAGCAGACCC TATCATCTCT CTCGTAAACT GCCGTCAGAG 2941 TCGGTTTGGT TGGACGAACC TTCTGAGTTT CTGGTAACGC CGTTCCGCAC CCCGGAAATG 3001 GTCAGCGAAC CAATCAGCAG GGTCATCGCT AGCCAGATCC TCTACGCCGG ACGCATCGTG 3061 GCCGGCATCA CCGGCGCCAC AGGTGCGGTT GCTGGCGCCT ATATCGCCGA CATCACCGAT 3121 GGGGAAGATC GGGCTCGCCA CTTCGGGCTC ATGAGCGCTT GTTTCGGCGT GGGTATGGTG 3181 GCAGGCCCCG TGGCCGGGGG ACTGTTGGGC GCCATCTCCT TGCACCATTC CTTGCGGCGG 3241 CGGTGCTCAA CGGCCTCAAC CTACTACTGG GCTGCTTCCT AATGCAGGAG TCGCATAAGG 3301 GAGAGCGTCG ATATGGTGCA CTCTCAGTAC AATCTGCTCT GATGCCGCAT AGTTAAGCCA 3361 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC 3421 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC 3481 ATCACCGAAA CGCGCGAGAC GAAAGGGCCT CGTGATACGC CTATTTTTAT AGGTTAATGT 3541 CATGATAATA ATGGTTTCTT AGACGTCAGG TGGCACTTTT CGGGGAAATG TGCGCGGAAC 3601 CCCTATTTGT TTATTTTTCT AAATACATTC AAATATGTAT CCGCTCATGA GACAATAACC 3661 CTGATAAATG CTTCAATAAT ATTGAAAAAG GAAGAGTATG AGTATTCAAC ATTTCCGTGT 3721 CGCCCTTATT CCCTTTTTTG CGGCATTTTG CCTTCCTGTT TTTGCTCACC CAGAAACGCT 3781 GGTGAAAGTA AAAGATGCTG AAGATCAGTT GGGTGCACGA GTGGGTTACA TCGAACTGGA 3841 TCTCAACAGC GGTAAGATCC TTGAGAGTTT TCGCCCCGAA GAACGTTTTC CAATGATGAG 3901 CACTTTTAAA GTTCTGCTAT GTGGCGCGGT ATTATCCCGT ATTGACGCCG GGCAAGAGCA 3961 ACTCGGTCGC CGCATACACT ATTCTCAGAA TGACTTGGTT GAGTACTCAC CAGTCACAGA 4021 AAAGCATCTT ACGGATGGCA TGACAGTAAG AGAATTATGC AGTGCTGCCA TAACCATGAG 4081 TGATAACACT GCGGCCAACT TACTTCTGAC AACGATCGGA GGACCGAAGG AGCTAACCGC 4141 TTTTTTGCAC AACATGGGGG ATCATGTAAC TCGCCTTGAT CGTTGGGAAC CGGAGCTGAA 4201 TGAAGCCATA CCAAACGACG AGCGTGACAC CACGATGCCT GTAGCAATGG CAACAACGTT 4261 GCGCAAACTA TTAACTGGCG AACTACTTAC TCTAGCTTCC CGGCAACAAT TAATAGACTG 4321 GATGGAGGCG GATAAAGTTG CAGGACCACT TCTGCGCTCG GCCCTTCCGG CTGGCTGGTT 4381 TATTGCTGAT AAATCTGGAG CCGGTGAGCG TGGGTCTCGC GGTATCATTG CAGCACTGGG 4441 GCCAGATGGT AAGCCCTCCC GTATCGTAGT TATCTACACG ACGGGGAGTC AGGCAACTAT 4501 GGATGAACGA AATAGACAGA TCGCTGAGAT AGGTGCCTCA CTGATTAAGC ATTGGTAACT 4561 GTCAGACCAA GTTTACTCAT ATATACTTTA GATTGATTTA AAACTTCATT TTTAATTTAA 4621 AAGGATCTAG GTGAAGATCC TTTTTGATAA TCTCATGACC AAAATCCCTT AACGTGAGTT 4681 TTCGTTCCAC TGAGCGTCAG ACCCCGTAGA AAAGATCAAA GGATCTTCTT GAGATCCTTT 4741 TTTTCTGCGC GTAATCTGCT GCTTGCAAAC AAAAAAACCA CCGCTACCAG CGGTGGTTTG 4801 TTTGCCGGAT CAAGAGCTAC CAACTCTTTT TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA 4861 GATACCAAAT ACTGTCCTTC TAGTGTAGCC GTAGTTAGGC CACCACTTCA AGAACTCTGT 4921 AGCACCGCCT ACATACCTCG CTCTGCTAAT CCTGTTACCA GTGGCTGCTG CCAGTGGCGA 4981 TAAGTCGTGT CTTACCGGGT TGGACTCAAG ACGATAGTTA CCGGATAAGG CGCAGCGGTC 5041 GGGCTGAACG GGGGGTTCGT GCACACAGCC CAGCTTGGAG CGAACGACCT ACACCGAACT 5101 GAGATACCTA CAGCGTGAGC TATGAGAAAG CGCCACGCTT CCCGAAGGGA GAAAGGCGGA 5161 CAGGTATCCG GTAAGCGGCA GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG 5221 AAACGCCTGG TATCTTTATA GTCCTGTCGG GTTTCGCCAC CTCTGACTTG AGCGTCGATT 5281 TTTGTGATGC TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC GCCAGCAACG CGGCCTTTTT 5341 ACGGTTCCTG GCCTTTTGCT GGCCTTTTGC TCACATGTTC TTTCCTGCGT TATCCCCTGA 5401 TTCTGTGGAT AACCGTATTA CCGCCTTTGA GTGAGCTGAT ACCGCTCGCC GCAGCCGAAC 5461 GACCGAGCGC AGCGAGTCAG TGAGCGAGGA AGCGGAAGAG CGCCCAATAC GCAAACCGCC 5521 TCTCCCCGCG CGTTGGCCGA TTCATTAATG CAG //