LOCUS pyrFEKO-NEO 6018 bp DNA circular 11-MAY-2010 The two PvuII sites that flank the PYR-FE targeting sequences are not shown on the map. FEATURES Location/Qualifiers misc_feature 2662..3158 /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 4163..5020 /function="LACTAMASE" misc_feature 4..365 /note="PYR-FE upstream" /note="ends 136 bp before start of CDS." protein_bind 372..405 /function="loxP" 5'UTR 433..518 /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 519..576 /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." CDS 1449..1478 /note="Ty-1 epitope" CDS 1485..2615 /note="HSVTK" 3'UTR 3167..3225 /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." protein_bind 2622..2655 /function="loxP" CDS 579..1442 /function="NEO" CDS 579..2615 /note="NEO-TK" misc_feature 649..669 /note="ADDGENE SEQ ERROR deletes next T making frameshift. This was checked by us and not so: sequence is fine." misc_feature join(5900..6018,1..1120) /note="verified sequence" /note="Us and Addgene March 2010." ORIGIN 1 CTGTGGCGAA TAATTGTAGC GGCAGTTTCT AACGGTTATA TGTTTAACTG CATTATGCCC 61 CTCTCTCACT TGGCGCTATC TCTGTGTTAC AACATGCGTT TGATCTATCA GTTTATTATA 121 TTGTTTTCCC TCAAACACAT GCGAAGCGAG TAAGAGTTGA AAATGTTACG GCAAAGGCCA 181 ATGGGCACTG GTAGAAGGCA ACGGATCGCG GTTATGAGAG TTCTTGTGTA TATGTTTATG 241 TGGGGGTGGG GGTGGAAGTT GTGGTAGTGT CATCTCGTGT ACATTGTTAA ACTCGCTTCA 301 ACCTTGTTTC TTTTTTCCTT TTCTTCTTCT TAGCTATCTG ATTGAGTTTT CATACTAAGT 361 TGGATAAGCT TATAACTTCG TATAGCATAC ATTATACGAA GTTATaccgg tACAcGTTCT 421 CGTCggccgg ccGCTGCACG CGCCTTCAAG TTTTTTTTCC TTTTCCCCAT TTTTTTCAAC 481 TTGAAGaCTT CAATTACACC AAAAAATAAA ATTCACAAAC TTGGtATTCC TTTGTGTTAC 541 ATTCTTGATC GCTCGCACTG ACATTACaga tctcgaatat gcgcgaaatc gtctgcgttc 601 aggctggcca atgcggtaac cagatcggct caaagttctg ggaggtgatc cggccaagtt 661 tggatggatT gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact 721 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc 781 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg 841 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagccgtg ctcgacgttg 901 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt 961 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc 1021 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag 1081 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg 1141 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc 1201 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt 1261 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg 1321 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt 1381 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct 1441 tcTCTAGAGA AGTCCATACT AACCAGGACC CACTTGACGC CACCATGGCC TCGTACCCCG 1501 GCCATCAACA CGCGTCTGCG TTCGACCAGG CTGCGCGTTC TCGCGGCCAT AGCAACCGAC 1561 GTACGGCGTT GCGCCCTCGC CGGCAGCAAG AAGCCACGGA AGTCCGCCCG GAGCAGAAAA 1621 TGCCCACGCT ACTGCGGGTT TATATAGACG GTCCCCACGG GATGGGGAAA ACCACCACCA 1681 CGCAACTGCT GGTGGCCCTG GGTTCGCGCG ACGATATCGT CTACGTACCC GAGCCGATGA 1741 CTTACTGGCG GGTGCTGGGG GCTTCCGAGA CAATCGCGAA CATCTACACC ACACAACACC 1801 GCCTCGACCA GGGTGAGATA TCGGCCGGGG ACGCGGCGGT GGTAATGACA AGCGCCCAGA 1861 TAACAATGGG CATGCCTTAT GCCGTGACCG ACGCCGTTCT GGCTCCTCAT ATCGGGGGGG 1921 AGGCTGGGAG CTCACATGCC CCGCCCCCGG CCCTCACCCT CATCTTCGAC CGCCATCCCA 1981 TCGCCGCCCT CCTGTGCTAC CCGGCCGCGC GGTACCTTAT GGGCAGCATG ACCCCCCAGG 2041 CCGTGCTGGC GTTCGTGGCC CTCATCCCGC CGACCTTGCC CGGCACCAAC ATCGTGCTTG 2101 GGGCCCTTCC GGAGGACAGA CACATCGACC GCCTGGCCAA ACGCCAGCGC CCCGGCGAGC 2161 GGCTGGACCT GGCTATGCTG GCTGCGATTC GCCGCGTTTA CGGGCTACTT GCCAATACGG 2221 TGCGGTATCT GCAGTGCGGC GGGTCGTGGC GGGAGGACTG GGGACAGCTT TCGGGGACGG 2281 CCGTGCCGCC CCAGGGTGCC GAGCCCCAGA GCAACGCGGG CCCACGACCC CATATCGGGG 2341 ACACGTTATT TACCCTGTTT CGGGCCCCCG AGTTGCTGGC CCCCAACGGC GACCTGTATA 2401 ACGTGTTTGC CTGGGCCTTG GACGTCTTGG CCAAACGCCT CCGTTCCATG CACGTCTTTA 2461 TCCTGGATTA CGACCAATCG CCCGCCGGCT GCCGGGACGC CCTGCTGCAA CTTACCTCCG 2521 GGATGGTCCA GACCCACGTC ACCACCCCCG GCTCCATACC GACGATATGC GACCTGGCGC 2581 GCACGTTTGC CCGGGAGATG GGGGAGGCTA ACTGAGAATT CATAACTTCG TATAGCATAC 2641 ATTATACGAA GTTATGGATC CTAAGTGGGT CGGAGCTCTA TCTTTAAGCT ATTGCGGACG 2701 TACACACATG TTTTCGTTGC AAATTATTTT CGtACCTACT TCAGATCGTA AGCGTGGGGA 2761 ATAATAATAA TGCTCCCTAC CATGAATTTA AaCAGTTTGG TGAATGAACA ATTCaCTTTA 2821 AATTGATGGA CCATGAAATG CACTTTTACG GTCGCAGCGT TTAACTAAGT GGCGAaGGCA 2881 AGTTTTTAAT AATAATAATA AAGCAAgTAG TAAACTATAT GAGACAGCAA TGGGGTTTGG 2941 GAGGGAAGGT TTAATCGCTT CAAAGGTATT TGTGTGTGGT TGAGGAGGTG ATAGCGAAGT 3001 GAGGGTTTCT AATAACTGTA GAGCAGCAAT AAAAAAAAGG TAGCAGTTGA TCAATTTGCT 3061 GTGGTGCCCT CTGCATTGAA GGGTATCTGG GATGTATGAA GTTCCATCAC TCGGAGCCAT 3121 CACCTTTCCC TCATTTCTCG TTTACATCCT TTACATGTCA GCTGCCTGCA GGTTGGTTAG 3181 GAAGGGGGGA TGATGTAAAA GAAGAAAATG GGGGGATTCG AGCCCCCTTG CAGGCATGCA 3241 AGCTAGCTTG TATTCTATAG TGTCACCTAA ATCGTATGTG TATGATACAT AAGGTTATGT 3301 ATTAATTGTA GCCGCGTTCT AACGACAATA TGTACAAGCC TAATTGTGTA GCATCTGGCT 3361 TACTGAAGCA GACCCTATCA TCTCTCTCGT AAACTGCCGT CAGAGTCGGT TTGGTTGGAC 3421 GAACCTTCTG AGTTTCTGGT AACGCCGTTC CGCACCCCGG AAATGGTCAG CGAACCAATC 3481 AGCAGGGTCA TCGCTAGCCA GATCCTCTAC GCCGGACGCA TCGTGGCCGG CATCACCGGC 3541 GCCACAGGTG CGGTTGCTGG CGCCTATATC GCCGACATCA CCGATGGGGA AGATCGGGCT 3601 CGCCACTTCG GGCTCATGAG CGCTTGTTTC GGCGTGGGTA TGGTGGCAGG CCCCGTGGCC 3661 GGGGGACTGT TGGGCGCCAT CTCCTTGCAC CATTCCTTGC GGCGGCGGTG CTCAACGGCC 3721 TCAACCTACT ACTGGGCTGC TTCCTAATGC AGGAGTCGCA TAAGGGAGAG CGTCGATATG 3781 GTGCACTCTC AGTACAATCT GCTCTGATGC CGCATAGTTA AGCCAGCCCC GACACCCGCC 3841 AACACCCGCT GACGCGCCCT GACGGGCTTG TCTGCTCCCG GCATCCGCTT ACAGACAAGC 3901 TGTGACCGTC TCCGGGAGCT GCATGTGTCA GAGGTTTTCA CCGTCATCAC CGAAACGCGC 3961 GAGACGAAAG GGCCTCGTGA TACGCCTATT TTTATAGGTT AATGTCATGA TAATAATGGT 4021 TTCTTAGACG TCAGGTGGCA CTTTTCGGGG AAATGTGCGC GGAACCCCTA TTTGTTTATT 4081 TTTCTAAATA CATTCAAATA TGTATCCGCT CATGAGACAA TAACCCTGAT AAATGCTTCA 4141 ATAATATTGA AAAAGGAAGA GTATGAGTAT TCAACATTTC CGTGTCGCCC TTATTCCCTT 4201 TTTTGCGGCA TTTTGCCTTC CTGTTTTTGC TCACCCAGAA ACGCTGGTGA AAGTAAAAGA 4261 TGCTGAAGAT CAGTTGGGTG CACGAGTGGG TTACATCGAA CTGGATCTCA ACAGCGGTAA 4321 GATCCTTGAG AGTTTTCGCC CCGAAGAACG TTTTCCAATG ATGAGCACTT TTAAAGTTCT 4381 GCTATGTGGC GCGGTATTAT CCCGTATTGA CGCCGGGCAA GAGCAACTCG GTCGCCGCAT 4441 ACACTATTCT CAGAATGACT TGGTTGAGTA CTCACCAGTC ACAGAAAAGC ATCTTACGGA 4501 TGGCATGACA GTAAGAGAAT TATGCAGTGC TGCCATAACC ATGAGTGATA ACACTGCGGC 4561 CAACTTACTT CTGACAACGA TCGGAGGACC GAAGGAGCTA ACCGCTTTTT TGCACAACAT 4621 GGGGGATCAT GTAACTCGCC TTGATCGTTG GGAACCGGAG CTGAATGAAG CCATACCAAA 4681 CGACGAGCGT GACACCACGA TGCCTGTAGC AATGGCAACA ACGTTGCGCA AACTATTAAC 4741 TGGCGAACTA CTTACTCTAG CTTCCCGGCA ACAATTAATA GACTGGATGG AGGCGGATAA 4801 AGTTGCAGGA CCACTTCTGC GCTCGGCCCT TCCGGCTGGC TGGTTTATTG CTGATAAATC 4861 TGGAGCCGGT GAGCGTGGGT CTCGCGGTAT CATTGCAGCA CTGGGGCCAG ATGGTAAGCC 4921 CTCCCGTATC GTAGTTATCT ACACGACGGG GAGTCAGGCA ACTATGGATG AACGAAATAG 4981 ACAGATCGCT GAGATAGGTG CCTCACTGAT TAAGCATTGG TAACTGTCAG ACCAAGTTTA 5041 CTCATATATA CTTTAGATTG ATTTAAAACT TCATTTTTAA TTTAAAAGGA TCTAGGTGAA 5101 GATCCTTTTT GATAATCTCA TGACCAAAAT CCCTTAACGT GAGTTTTCGT TCCACTGAGC 5161 GTCAGACCCC GTAGAAAAGA TCAAAGGATC TTCTTGAGAT CCTTTTTTTC TGCGCGTAAT 5221 CTGCTGCTTG CAAACAAAAA AACCACCGCT ACCAGCGGTG GTTTGTTTGC CGGATCAAGA 5281 GCTACCAACT CTTTTTCCGA AGGTAACTGG CTTCAGCAGA GCGCAGATAC CAAATACTGT 5341 CCTTCTAGTG TAGCCGTAGT TAGGCCACCA CTTCAAGAAC TCTGTAGCAC CGCCTACATA 5401 CCTCGCTCTG CTAATCCTGT TACCAGTGGC TGCTGCCAGT GGCGATAAGT CGTGTCTTAC 5461 CGGGTTGGAC TCAAGACGAT AGTTACCGGA TAAGGCGCAG CGGTCGGGCT GAACGGGGGG 5521 TTCGTGCACA CAGCCCAGCT TGGAGCGAAC GACCTACACC GAACTGAGAT ACCTACAGCG 5581 TGAGCTATGA GAAAGCGCCA CGCTTCCCGA AGGGAGAAAG GCGGACAGGT ATCCGGTAAG 5641 CGGCAGGGTC GGAACAGGAG AGCGCACGAG GGAGCTTCCA GGGGGAAACG CCTGGTATCT 5701 TTATAGTCCT GTCGGGTTTC GCCACCTCTG ACTTGAGCGT CGATTTTTGT GATGCTCGTC 5761 AGGGGGGCGG AGCCTATGGA AAAACGCCAG CAACGCGGCC TTTTTACGGT TCCTGGCCTT 5821 TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC CCTGATTCTG TGGATAACCG 5881 TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC CGAACGACCG AGCGCAGCGA 5941 GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA CCGCCTCTCC CCGCGCGTTG 6001 GCCGATTCAT TAATGCAG //