LOCUS pyrFEKO-HYG 6191 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 2835..3331 /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 4336..5193 /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 3340..3398 /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 590..2785 /function="HYG-TK" CDS 1622..1651 /function="Ty-1 epitope" CDS 1658..2788 /note="HSVTK CDS" protein_bind 2795..2828 /function="loxP" CDS 590..1616 /note="HYG CDS" BASE COUNT 1405 a 1621 c 1653 g 1512 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 CagatctTAC TGTTGGTAAA TGATGAAAAA 601 GCCTGAACTC ACCGCGACGT CTGTCGAGAA GTTTCTGATC GAAAAGTTCG ACAGCGTCTC 661 CGACCTGATG CAGCTCTCGG AGGGCGAAGA ATCTCGTGCT TTCAGCTTCG ATGTAGGAGG 721 GCGTGGATAT GTCCTGCGGG TAAATAGCTG CGCCGATGGT TTCTACAAAG ATCGTTATGT 781 TTATCGGCAC TTTGCATCGG CCGCGCTCCC GATTCCGGAA GTGCTTGACA TTGGGGAATT 841 CAGCGAGAGC CTGACCTATT GCATCTCCCG CCGTGCACAG GGTGTCACGT TGCAAGACCT 901 GCCTGAAACC GAACTGCCCG CTGTTCTGCA GCCGGTCGCG GAGGCCATGG ATGCGATCGC 961 TGCGGCCGAT CTTAGCCAGA CGAGCGGGTT CGGCCCATTC GGACCGCAAG GAATCGGTCA 1021 ATACACTACA TGGCGTGATT TCATATGCGC GATTGCTGAT CCCCATGTGT ATCACTGGCA 1081 AACTGTGATG GACGACACCG TCAGTGCGTC CGTCGCGCAG GCTCTCGATG AGCTGATGCT 1141 TTGGGCCGAG GACTGCCCCG AAGTCCGGCA CCTCGTGCAC GCGGATTTCG GCTCCAACAA 1201 TGTCCTGACG GACAATGGCC GCATAACAGC GGTCATTGAC TGGAGCGAGG CGATGTTCGG 1261 GGATTCCCAA TACGAGGTCG CCAACATCTT CTTCTGGAGG CCGTGGTTGG CTTGTATGGA 1321 GCAGCAGACG CGCTACTTCG AGCGGAGGCA TCCGGAGCTT GCAGGATCGC CGCGGCTCCG 1381 GGCGTATATG CTCCGCATTG GTCTTGACCA ACTCTATCAG AGCTTGGTTG ACGGCAATTT 1441 CGATGATGCA GCTTGGGCGC AGGGTCGATG CGACGCAATC GTCCGATCCG GAGCCGGGAC 1501 TGTCGGGCGT ACACAAATCG CCCGCAGAAG CGCGGCCGTC TGGACCGATG GCTGTGTAGA 1561 AGTACTCGCC GATAGTGGAA ACCGACGCCC CAGCACTCGT CCGAGGGCAA AGGAATCTAG 1621 AGAAGTCCAT ACTAACCAGG ACCCACTTGA CGCCACCATG GCCTCGTACC CCGGCCATCA 1681 ACACGCGTCT GCGTTCGACC AGGCTGCGCG TTCTCGCGGC CATAGCAACC GACGTACGGC 1741 GTTGCGCCCT CGCCGGCAGC AAGAAGCCAC GGAAGTCCGC CCGGAGCAGA AAATGCCCAC 1801 GCTACTGCGG GTTTATATAG ACGGTCCCCA CGGGATGGGG AAAACCACCA CCACGCAACT 1861 GCTGGTGGCC CTGGGTTCGC GCGACGATAT CGTCTACGTA CCCGAGCCGA TGACTTACTG 1921 GCGGGTGCTG GGGGCTTCCG AGACAATCGC GAACATCTAC ACCACACAAC ACCGCCTCGA 1981 CCAGGGTGAG ATATCGGCCG GGGACGCGGC GGTGGTAATG ACAAGCGCCC AGATAACAAT 2041 GGGCATGCCT TATGCCGTGA CCGACGCCGT TCTGGCTCCT CATATCGGGG GGGAGGCTGG 2101 GAGCTCACAT GCCCCGCCCC CGGCCCTCAC CCTCATCTTC GACCGCCATC CCATCGCCGC 2161 CCTCCTGTGC TACCCGGCCG CGCGGTACCT TATGGGCAGC ATGACCCCCC AGGCCGTGCT 2221 GGCGTTCGTG GCCCTCATCC CGCCGACCTT GCCCGGCACC AACATCGTGC TTGGGGCCCT 2281 TCCGGAGGAC AGACACATCG ACCGCCTGGC CAAACGCCAG CGCCCCGGCG AGCGGCTGGA 2341 CCTGGCTATG CTGGCTGCGA TTCGCCGCGT TTACGGGCTA CTTGCCAATA CGGTGCGGTA 2401 TCTGCAGTGC GGCGGGTCGT GGCGGGAGGA CTGGGGACAG CTTTCGGGGA CGGCCGTGCC 2461 GCCCCAGGGT GCCGAGCCCC AGAGCAACGC GGGCCCACGA CCCCATATCG GGGACACGTT 2521 ATTTACCCTG TTTCGGGCCC CCGAGTTGCT GGCCCCCAAC GGCGACCTGT ATAACGTGTT 2581 TGCCTGGGCC TTGGACGTCT TGGCCAAACG CCTCCGTTCC ATGCACGTCT TTATCCTGGA 2641 TTACGACCAA TCGCCCGCCG GCTGCCGGGA CGCCCTGCTG CAACTTACCT CCGGGATGGT 2701 CCAGACCCAC GTCACCACCC CCGGCTCCAT ACCGACGATA TGCGACCTGG CGCGCACGTT 2761 TGCCCGGGAG ATGGGGGAGG CTAACTGAGA ATTCATAACT TCGTATAGCA TACATTATAC 2821 GAAGTTATGG ATCCTAAGTG GGTCGGAGCT CTATCTTTAA GCTATTGCGG ACGTACACAC 2881 ATGTTTTCGT TGCAAATTAT TTTCGtACCT ACTTCAGATC GTAAGCGTGG GGAATAATAA 2941 TAATGCTCCC TACCATGAAT TTAAaCAGTT TGGTGAATGA ACAATTCaCT TTAAATTGAT 3001 GGACCATGAA ATGCACTTTT ACGGTCGCAG CGTTTAACTA AGTGGCGAaG GCAAGTTTTT 3061 AATAATAATA ATAAAGCAAg TAGTAAACTA TATGAGACAG CAATGGGGTT TGGGAGGGAA 3121 GGTTTAATCG CTTCAAAGGT ATTTGTGTGT GGTTGAGGAG GTGATAGCGA AGTGAGGGTT 3181 TCTAATAACT GTAGAGCAGC AATAAAAAAA AGGTAGCAGT TGATCAATTT GCTGTGGTGC 3241 CCTCTGCATT GAAGGGTATC TGGGATGTAT GAAGTTCCAT CACTCGGAGC CATCACCTTT 3301 CCCTCATTTC TCGTTTACAT CCTTTACATG TCAGCTGCCT GCAGGTTGGT TAGGAAGGGG 3361 GGATGATGTA AAAGAAGAAA ATGGGGGGAT TCGAGCCCCC TTGCAGGCAT GCAAGCTAGC 3421 TTGTATTCTA TAGTGTCACC TAAATCGTAT GTGTATGATA CATAAGGTTA TGTATTAATT 3481 GTAGCCGCGT TCTAACGACA ATATGTACAA GCCTAATTGT GTAGCATCTG GCTTACTGAA 3541 GCAGACCCTA TCATCTCTCT CGTAAACTGC CGTCAGAGTC GGTTTGGTTG GACGAACCTT 3601 CTGAGTTTCT GGTAACGCCG TTCCGCACCC CGGAAATGGT CAGCGAACCA ATCAGCAGGG 3661 TCATCGCTAG CCAGATCCTC TACGCCGGAC GCATCGTGGC CGGCATCACC GGCGCCACAG 3721 GTGCGGTTGC TGGCGCCTAT ATCGCCGACA TCACCGATGG GGAAGATCGG GCTCGCCACT 3781 TCGGGCTCAT GAGCGCTTGT TTCGGCGTGG GTATGGTGGC AGGCCCCGTG GCCGGGGGAC 3841 TGTTGGGCGC CATCTCCTTG CACCATTCCT TGCGGCGGCG GTGCTCAACG GCCTCAACCT 3901 ACTACTGGGC TGCTTCCTAA TGCAGGAGTC GCATAAGGGA GAGCGTCGAT ATGGTGCACT 3961 CTCAGTACAA TCTGCTCTGA TGCCGCATAG TTAAGCCAGC CCCGACACCC GCCAACACCC 4021 GCTGACGCGC CCTGACGGGC TTGTCTGCTC CCGGCATCCG CTTACAGACA AGCTGTGACC 4081 GTCTCCGGGA GCTGCATGTG TCAGAGGTTT TCACCGTCAT CACCGAAACG CGCGAGACGA 4141 AAGGGCCTCG TGATACGCCT ATTTTTATAG GTTAATGTCA TGATAATAAT GGTTTCTTAG 4201 ACGTCAGGTG GCACTTTTCG GGGAAATGTG CGCGGAACCC CTATTTGTTT ATTTTTCTAA 4261 ATACATTCAA ATATGTATCC GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT 4321 TGAAAAAGGA AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG 4381 GCATTTTGCC TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA AGATGCTGAA 4441 GATCAGTTGG GTGCACGAGT GGGTTACATC GAACTGGATC TCAACAGCGG TAAGATCCTT 4501 GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA ATGATGAGCA CTTTTAAAGT TCTGCTATGT 4561 GGCGCGGTAT TATCCCGTAT TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT 4621 TCTCAGAATG ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG 4681 ACAGTAAGAG AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC GGCCAACTTA 4741 CTTCTGACAA CGATCGGAGG ACCGAAGGAG CTAACCGCTT TTTTGCACAA CATGGGGGAT 4801 CATGTAACTC GCCTTGATCG TTGGGAACCG GAGCTGAATG AAGCCATACC AAACGACGAG 4861 CGTGACACCA CGATGCCTGT AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA 4921 CTACTTACTC TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA 4981 GGACCACTTC TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA ATCTGGAGCC 5041 GGTGAGCGTG GGTCTCGCGG TATCATTGCA GCACTGGGGC CAGATGGTAA GCCCTCCCGT 5101 ATCGTAGTTA TCTACACGAC GGGGAGTCAG GCAACTATGG ATGAACGAAA TAGACAGATC 5161 GCTGAGATAG GTGCCTCACT GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT 5221 ATACTTTAGA TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT 5281 TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC 5341 CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC 5401 TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA 5461 ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA 5521 GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT 5581 CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG 5641 GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC 5701 ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA 5761 TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG 5821 GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT 5881 CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG 5941 CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG 6001 CCTTTTGCTC ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC 6061 GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG 6121 AGCGAGGAAG CGGAAGAGCG CCCAATACGC AAACCGCCTC TCCCCGCGCG TTGGCCGATT 6181 CATTAATGCA G //