LOCUS pMOTag4HM-crelox-PUR 5188 bp DNA circular 11-APR-2009 pMOTag4HM-crelox-PUR made by Jenny Li taking the HindIII-BamHI crelox-PUR fragment from pyrFEKO-PUR and inserting into HindIII-BamHI digested pMOTag4HM made by Jessica. Oberholzer,M., Morand,S., Kunz,S., and Seebeck,T.  A vector series for rapid PCR-mediated C-terminal in-situ tagging of T. brucei genes. Mol. Biochem. Parasitol. 145, 2005, 117-120. Sequence file compiled April 2009 by George From actual sequence, made 3 corrections. GCGC>tcga at 3019-3022; T>G at 2736 and G>T at 2763, both within HA tag but no AA change. FEATURES Location/Qualifiers rep_origin complement(135..440) /note="f1 origin of ss-DNA replication" promoter 625..644 /note="T7 Promoter" promoter complement(2999..3018) /note="T3 Promoter" rep_origin 3485..4052 /note="pUC origin" CDS complement(4200..5060) /note="AMP" promoter complement(3044..3165) /note="lac Promoter" CDS complement(2805..2963) /gene="3xcMyc" /translation="GGRSRSEEQKLISEEDLLRSEEQKLISEEDLLRSEEQKLISEED LLRSEAAVE" /note="LRSEAAVE linker added before HIS tag" CDS complement(2724..2804) /gene="3xHA" /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" CDS complement(735..1865) /note="HSVTK" misc_signal complement(694..728) /function="loxP" CDS complement(1908..2504) /function="PUR" /note="Corrected sequence position 1935 a>g and 2118 g>a. Probably same as noted elsewhere in these plasmid PUR sequences, and does not change the amino acids." CDS complement(1872..1901) /function="Ty-1 epitope" misc_signal complement(2676..2709) /function="loxP" 5'UTR complement(2563..2648) /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)" misc_feature complement(2505..2562) /function="Residue from original LUC insert" /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." misc_feature 629..1370 /note="100% verified sequence" misc_feature 2282..3067 /note="100% verified sequence" BASE COUNT 1146 a 1389 c 1512 g 1141 t ORIGIN 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca 661 ccgcggtggc ggccgctcta gaactagtgg atccATAACT TCGTATAATG TATGCTATAC 721 GAAGTTATGA ATTCTCAGTT AGCCTCCCCC ATCTCCCGGG CAAACGTGCG CGCCAGGTCG 781 CATATCGTCG GTATGGAGCC GGGGGTGGTG ACGTGGGTCT GGACCATCCC GGAGGTAAGT 841 TGCAGCAGGG CGTCCCGGCA GCCGGCGGGC GATTGGTCGT AATCCAGGAT AAAGACGTGC 901 ATGGAACGGA GGCGTTTGGC CAAGACGTCC AAGGCCCAGG CAAACACGTT ATACAGGTCG 961 CCGTTGGGGG CCAGCAACTC GGGGGCCCGA AACAGGGTAA ATAACGTGTC CCCGATATGG 1021 GGTCGTGGGC CCGCGTTGCT CTGGGGCTCG GCACCCTGGG GCGGCACGGC CGTCCCCGAA 1081 AGCTGTCCCC AGTCCTCCCG CCACGACCCG CCGCACTGCA GATACCGCAC CGTATTGGCA 1141 AGTAGCCCGT AAACGCGGCG AATCGCAGCC AGCATAGCCA GGTCCAGCCG CTCGCCGGGG 1201 CGCTGGCGTT TGGCCAGGCG GTCGATGTGT CTGTCCTCCG GAAGGGCCCC AAGCACGATG 1261 TTGGTGCCGG GCAAGGTCGG CGGGATGAGG GCCACGAACG CCAGCACGGC CTGGGGGGTC 1321 ATGCTGCCCA TAAGGTACCG CGCGGCCGGG TAGCACAGGA GGGCGGCGAT GGGATGGCGG 1381 TCGAAGATGA GGGTGAGGGC CGGGGGCGGG GCATGTGAGC TCCCAGCCTC CCCCCCGATA 1441 TGAGGAGCCA GAACGGCGTC GGTCACGGCA TAAGGCATGC CCATTGTTAT CTGGGCGCTT 1501 GTCATTACCA CCGCCGCGTC CCCGGCCGAT ATCTCACCCT GGTCGAGGCG GTGTTGTGTG 1561 GTGTAGATGT TCGCGATTGT CTCGGAAGCC CCCAGCACCC GCCAGTAAGT CATCGGCTCG 1621 GGTACGTAGA CGATATCGTC GCGCGAACCC AGGGCCACCA GCAGTTGCGT GGTGGTGGTT 1681 TTCCCCATCC CGTGGGGACC GTCTATATAA ACCCGCAGTA GCGTGGGCAT TTTCTGCTCC 1741 GGGCGGACTT CCGTGGCTTC TTGCTGCCGG CGAGGGCGCA ACGCCGTACG TCGGTTGCTA 1801 TGGCCGCGAG AACGCGCAGC CTGGTCGAAC GCAGACGCGT GTTGATGGCC GGGGTACGAG 1861 GCCATGGTGG CGTCAAGTGG GTCCTGGTTA GTATGGACTT CTCTAGAGGC ACCGGGCTTG 1921 CGGGTCATGC ACCAgGTGCG CGGTCCTTCG GGCACCTCGA CGTCGGCGGT GACGGTGAAG 1981 CCGAGCCGCT CGTAGAAGGG GAGGTTGCGG GGCGCGGAGG TCTCCAGGAA GGCGGGCACC 2041 CCGGCGCGCT CGGCCGCCTC CACTCCGGGG AGCACGACGG CGCTGCCCAG ACCCTTGCCC 2101 TGGTGGTCGG GCGAGACaCC GACGGTGGCC AGGAACCACG CGGGCTCCTT GGGCCGGTGC 2161 GGCGCCAGGA GGCCTTCCAT CTGTTGCTGC GCGGCCAGCC GGGAACCGCT CAACTCGGCC 2221 ATGCGCGGGC CGATCTCGGC GAACACCGCC CCCGCTTCGA CGCTCTCCGG CGTGGTCCAG 2281 ACCGCCACTG CTGCGCCGTC GTCCGCGACC CACACCTTGC CGATGTCGAG CCCGACGCGC 2341 GTGAGGAAGA GTTCTTGCAG CTCGGTGACC CGCTCGATGT GGCGGTCTGG ATCGACGGTG 2401 TGGCGCGTGG CGGGGTAGTC GGCGAACGCG GCGGCGAGGG TGCGTACGGC CCTGGGGACG 2461 TCGTCGCGGG TGGCGAGGCG CACCGTGGGC TTGTACTCGG TCATGGTAGA TCtGTAATGT 2521 CAGTGCGAGC GATCAAGAAT GTAACACAAA GGAATaCCAA GTTTGTGAAT TTTATTTTTT 2581 GGTGTAATTG AAGtCTTCAA GTTGAAAAAA ATGGGGAAAA GGAAAAAAAA CTCGAAGGCG 2641 CGTGCAGCgg ccggccGACG AGAACgTGTa ccggtATAAC TTCGTATAAT GTATGCTATA 2701 CGAAGTTATA AGCTTATCGT CGACTATGCG TAATCgGGCA CATCGTACGG GTATGCGTAG 2761 TCtGGCACGT CGTATGGGTA CGCGTAATCA GGCACATCGT AAGGGTActc gacGGCCGCT 2821 TCAGATCGCA GCAGATCCTC CTCTGAGATC AGCTTCTGCT CCTCGGATCT CAGCAGATCC 2881 TCCTCCGAGA TCAGCTTCTG CTCCTCGGAT CTCAGCAGAT CCTCCTCCGA GATCAGCTTC 2941 TGCTCCTCGG ATCTCGATCG GCCGCCctcg agggggggcc cggtacccag cttttgttcc 3001 ctttagtgag ggttaattTC GAgcttggcg taatcatggt catagctgtt tcctgtgtga 3061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 3121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 3181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 3241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 3301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 3361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 3421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 3481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 3541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 3601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 3661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 3721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 3781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 3841 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc 3901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 3961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 4141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 4201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 4261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 4321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 4381 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 4441 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 4501 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 4561 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 4621 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 4681 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 4741 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 4801 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 4861 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 4921 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 4981 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 5041 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 5101 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 5161 ccgcgcacat ttccccgaaa agtgccac //