LOCUS pyrFEKO-BLE 5529 bp DNA circular 17-JUL-2008 The two PvuII sites that flank the PYR-FE targeting sequences are not shown on the map. FEATURES Location/Qualifiers misc_feature 2173..2669 /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 3674..4531 /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 2678..2736 /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..953 /note="BLE" /note="Phleomycin resistance" CDS 996..2126 /note="HSVTK" CDS 960..989 /function="Ty-1 epitope" CDS 582..2126 /function="BLE-TK" protein_bind 2133..2166 /function="loxP" BASE COUNT 1243 a 1459 c 1479 g 1348 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 CagatctCAC CATGGCCAAG TTGACCAGTG 601 CCGTTCCGGT GCTCACCGCG CGCGACGTCG CCGGAGCGGT CGAGTTCTGG ACCGACCGGC 661 TCGGGTTCTC CCGGGACTTC GTGGAGGACG ACTTCGCCGG TGTGGTCCGG GACGACGTGA 721 CCCTGTTCAT CAGCGCGGTC CAGGACCAGG TGGTGCCGGA CAACACCCTG GCCTGGGTGT 781 GGGTGCGCGG CCTGGACGAG CTGTACGCCG AGTGGTCGGA GGTCGTGTCC ACGAACTTCC 841 GGGACGCCTC CGGGCCGGCC ATGACCGAGA TCGGCGAGCA GCCGTGGGGG CGGGAGTTCG 901 CCCTGCGCGA CCCGGCCGGC AACTGCGTGC ACTTCGTGGC CGAGGAGCAG GACTCTAGAG 961 AAGTCCATAC TAACCAGGAC CCACTTGACG CCACCATGGC CTCGTACCCC GGCCATCAAC 1021 ACGCGTCTGC GTTCGACCAG GCTGCGCGTT CTCGCGGCCA TAGCAACCGA CGTACGGCGT 1081 TGCGCCCTCG CCGGCAGCAA GAAGCCACGG AAGTCCGCCC GGAGCAGAAA ATGCCCACGC 1141 TACTGCGGGT TTATATAGAC GGTCCCCACG GGATGGGGAA AACCACCACC ACGCAACTGC 1201 TGGTGGCCCT GGGTTCGCGC GACGATATCG TCTACGTACC CGAGCCGATG ACTTACTGGC 1261 GGGTGCTGGG GGCTTCCGAG ACAATCGCGA ACATCTACAC CACACAACAC CGCCTCGACC 1321 AGGGTGAGAT ATCGGCCGGG GACGCGGCGG TGGTAATGAC AAGCGCCCAG ATAACAATGG 1381 GCATGCCTTA TGCCGTGACC GACGCCGTTC TGGCTCCTCA TATCGGGGGG GAGGCTGGGA 1441 GCTCACATGC CCCGCCCCCG GCCCTCACCC TCATCTTCGA CCGCCATCCC ATCGCCGCCC 1501 TCCTGTGCTA CCCGGCCGCG CGGTACCTTA TGGGCAGCAT GACCCCCCAG GCCGTGCTGG 1561 CGTTCGTGGC CCTCATCCCG CCGACCTTGC CCGGCACCAA CATCGTGCTT GGGGCCCTTC 1621 CGGAGGACAG ACACATCGAC CGCCTGGCCA AACGCCAGCG CCCCGGCGAG CGGCTGGACC 1681 TGGCTATGCT GGCTGCGATT CGCCGCGTTT ACGGGCTACT TGCCAATACG GTGCGGTATC 1741 TGCAGTGCGG CGGGTCGTGG CGGGAGGACT GGGGACAGCT TTCGGGGACG GCCGTGCCGC 1801 CCCAGGGTGC CGAGCCCCAG AGCAACGCGG GCCCACGACC CCATATCGGG GACACGTTAT 1861 TTACCCTGTT TCGGGCCCCC GAGTTGCTGG CCCCCAACGG CGACCTGTAT AACGTGTTTG 1921 CCTGGGCCTT GGACGTCTTG GCCAAACGCC TCCGTTCCAT GCACGTCTTT ATCCTGGATT 1981 ACGACCAATC GCCCGCCGGC TGCCGGGACG CCCTGCTGCA ACTTACCTCC GGGATGGTCC 2041 AGACCCACGT CACCACCCCC GGCTCCATAC CGACGATATG CGACCTGGCG CGCACGTTTG 2101 CCCGGGAGAT GGGGGAGGCT AACTGAGAAT TCATAACTTC GTATAGCATA CATTATACGA 2161 AGTTATGGAT CCTAAGTGGG TCGGAGCTCT ATCTTTAAGC TATTGCGGAC GTACACACAT 2221 GTTTTCGTTG CAAATTATTT TCGtACCTAC TTCAGATCGT AAGCGTGGGG AATAATAATA 2281 ATGCTCCCTA CCATGAATTT AAaCAGTTTG GTGAATGAAC AATTCaCTTT AAATTGATGG 2341 ACCATGAAAT GCACTTTTAC GGTCGCAGCG TTTAACTAAG TGGCGAaGGC AAGTTTTTAA 2401 TAATAATAAT AAAGCAAgTA GTAAACTATA TGAGACAGCA ATGGGGTTTG GGAGGGAAGG 2461 TTTAATCGCT TCAAAGGTAT TTGTGTGTGG TTGAGGAGGT GATAGCGAAG TGAGGGTTTC 2521 TAATAACTGT AGAGCAGCAA TAAAAAAAAG GTAGCAGTTG ATCAATTTGC TGTGGTGCCC 2581 TCTGCATTGA AGGGTATCTG GGATGTATGA AGTTCCATCA CTCGGAGCCA TCACCTTTCC 2641 CTCATTTCTC GTTTACATCC TTTACATGTC AGCTGCCTGC AGGTTGGTTA GGAAGGGGGG 2701 ATGATGTAAA AGAAGAAAAT GGGGGGATTC GAGCCCCCTT GCAGGCATGC AAGCTAGCTT 2761 GTATTCTATA GTGTCACCTA AATCGTATGT GTATGATACA TAAGGTTATG TATTAATTGT 2821 AGCCGCGTTC TAACGACAAT ATGTACAAGC CTAATTGTGT AGCATCTGGC TTACTGAAGC 2881 AGACCCTATC ATCTCTCTCG TAAACTGCCG TCAGAGTCGG TTTGGTTGGA CGAACCTTCT 2941 GAGTTTCTGG TAACGCCGTT CCGCACCCCG GAAATGGTCA GCGAACCAAT CAGCAGGGTC 3001 ATCGCTAGCC AGATCCTCTA CGCCGGACGC ATCGTGGCCG GCATCACCGG CGCCACAGGT 3061 GCGGTTGCTG GCGCCTATAT CGCCGACATC ACCGATGGGG AAGATCGGGC TCGCCACTTC 3121 GGGCTCATGA GCGCTTGTTT CGGCGTGGGT ATGGTGGCAG GCCCCGTGGC CGGGGGACTG 3181 TTGGGCGCCA TCTCCTTGCA CCATTCCTTG CGGCGGCGGT GCTCAACGGC CTCAACCTAC 3241 TACTGGGCTG CTTCCTAATG CAGGAGTCGC ATAAGGGAGA GCGTCGATAT GGTGCACTCT 3301 CAGTACAATC TGCTCTGATG CCGCATAGTT AAGCCAGCCC CGACACCCGC CAACACCCGC 3361 TGACGCGCCC TGACGGGCTT GTCTGCTCCC GGCATCCGCT TACAGACAAG CTGTGACCGT 3421 CTCCGGGAGC TGCATGTGTC AGAGGTTTTC ACCGTCATCA CCGAAACGCG CGAGACGAAA 3481 GGGCCTCGTG ATACGCCTAT TTTTATAGGT TAATGTCATG ATAATAATGG TTTCTTAGAC 3541 GTCAGGTGGC ACTTTTCGGG GAAATGTGCG CGGAACCCCT ATTTGTTTAT TTTTCTAAAT 3601 ACATTCAAAT ATGTATCCGC TCATGAGACA ATAACCCTGA TAAATGCTTC AATAATATTG 3661 AAAAAGGAAG AGTATGAGTA TTCAACATTT CCGTGTCGCC CTTATTCCCT TTTTTGCGGC 3721 ATTTTGCCTT CCTGTTTTTG CTCACCCAGA AACGCTGGTG AAAGTAAAAG ATGCTGAAGA 3781 TCAGTTGGGT GCACGAGTGG GTTACATCGA ACTGGATCTC AACAGCGGTA AGATCCTTGA 3841 GAGTTTTCGC CCCGAAGAAC GTTTTCCAAT GATGAGCACT TTTAAAGTTC TGCTATGTGG 3901 CGCGGTATTA TCCCGTATTG ACGCCGGGCA AGAGCAACTC GGTCGCCGCA TACACTATTC 3961 TCAGAATGAC TTGGTTGAGT ACTCACCAGT CACAGAAAAG CATCTTACGG ATGGCATGAC 4021 AGTAAGAGAA TTATGCAGTG CTGCCATAAC CATGAGTGAT AACACTGCGG CCAACTTACT 4081 TCTGACAACG ATCGGAGGAC CGAAGGAGCT AACCGCTTTT TTGCACAACA TGGGGGATCA 4141 TGTAACTCGC CTTGATCGTT GGGAACCGGA GCTGAATGAA GCCATACCAA ACGACGAGCG 4201 TGACACCACG ATGCCTGTAG CAATGGCAAC AACGTTGCGC AAACTATTAA CTGGCGAACT 4261 ACTTACTCTA GCTTCCCGGC AACAATTAAT AGACTGGATG GAGGCGGATA AAGTTGCAGG 4321 ACCACTTCTG CGCTCGGCCC TTCCGGCTGG CTGGTTTATT GCTGATAAAT CTGGAGCCGG 4381 TGAGCGTGGG TCTCGCGGTA TCATTGCAGC ACTGGGGCCA GATGGTAAGC CCTCCCGTAT 4441 CGTAGTTATC TACACGACGG GGAGTCAGGC AACTATGGAT GAACGAAATA GACAGATCGC 4501 TGAGATAGGT GCCTCACTGA TTAAGCATTG GTAACTGTCA GACCAAGTTT ACTCATATAT 4561 ACTTTAGATT GATTTAAAAC TTCATTTTTA ATTTAAAAGG ATCTAGGTGA AGATCCTTTT 4621 TGATAATCTC ATGACCAAAA TCCCTTAACG TGAGTTTTCG TTCCACTGAG CGTCAGACCC 4681 CGTAGAAAAG ATCAAAGGAT CTTCTTGAGA TCCTTTTTTT CTGCGCGTAA TCTGCTGCTT 4741 GCAAACAAAA AAACCACCGC TACCAGCGGT GGTTTGTTTG CCGGATCAAG AGCTACCAAC 4801 TCTTTTTCCG AAGGTAACTG GCTTCAGCAG AGCGCAGATA CCAAATACTG TCCTTCTAGT 4861 GTAGCCGTAG TTAGGCCACC ACTTCAAGAA CTCTGTAGCA CCGCCTACAT ACCTCGCTCT 4921 GCTAATCCTG TTACCAGTGG CTGCTGCCAG TGGCGATAAG TCGTGTCTTA CCGGGTTGGA 4981 CTCAAGACGA TAGTTACCGG ATAAGGCGCA GCGGTCGGGC TGAACGGGGG GTTCGTGCAC 5041 ACAGCCCAGC TTGGAGCGAA CGACCTACAC CGAACTGAGA TACCTACAGC GTGAGCTATG 5101 AGAAAGCGCC ACGCTTCCCG AAGGGAGAAA GGCGGACAGG TATCCGGTAA GCGGCAGGGT 5161 CGGAACAGGA GAGCGCACGA GGGAGCTTCC AGGGGGAAAC GCCTGGTATC TTTATAGTCC 5221 TGTCGGGTTT CGCCACCTCT GACTTGAGCG TCGATTTTTG TGATGCTCGT CAGGGGGGCG 5281 GAGCCTATGG AAAAACGCCA GCAACGCGGC CTTTTTACGG TTCCTGGCCT TTTGCTGGCC 5341 TTTTGCTCAC ATGTTCTTTC CTGCGTTATC CCCTGATTCT GTGGATAACC GTATTACCGC 5401 CTTTGAGTGA GCTGATACCG CTCGCCGCAG CCGAACGACC GAGCGCAGCG AGTCAGTGAG 5461 CGAGGAAGCG GAAGAGCGCC CAATACGCAA ACCGCCTCTC CCCGCGCGTT GGCCGATTCA 5521 TTAATGCAG //