LOCUS pMOTag2T-crelox-HYG-SK+ 5467 bp DNA circular 18-MAR-2010 pMOTag4H-crelox-HYG made by Jenny Li taking the HindIII-BamHI crelox-HYG fragment from pyrFEKO-HYG and inserting into HindIII-BamHI digested pMOTag4H. Based on pMOTag series made in pBluescript II KS+ 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. FEATURES Location/Qualifiers rep_origin complement(135..440) /note="f1 origin of ss-DNA replication" promoter 625..644 /note="T7 Promoter" promoter complement(3278..3297) /note="T3 Promoter" rep_origin 3764..4331 /note="pUC origin" CDS complement(4479..5339) /note="AMP" promoter complement(3323..3444) /note="lac Promoter" CDS complement(3165..3245) /note="3xHA" misc_signal complement(3114..3147) /function="loxP" 5'UTR complement(3001..3086) /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(2934..3000) /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." CDS complement(1908..2933) /function="HYG" CDS complement(1872..1901) /function="Ty-1 epitope" CDS complement(735..1865) /note="HSVTK" misc_signal complement(695..728) /function="loxP" misc_feature 2255..3253 /note="Verified sequence" /note="Inserted an A in the sequence just before the Sal site. Last 670 bp also verified by Addgene, March 2010." 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 CTCTAGATTC CTTTGCCCTC 1921 GGACGAGTGC TGGGGCGTCG GTTTCCACTA TCGGCGAGTA CTTCTACACA GCCATCGGTC 1981 CAGACGGCCG CGCTTCTGCG GGCGATTTGT GTACGCCCGA CAGTCCCGGC TCCGGATCGG 2041 ACGATTGCGT CGCATCGACC CTGCGCCCAA GCTGCATCAT CGAAATTGCC GTCAACCAAG 2101 CTCTGATAGA GTTGGTCAAG ACCAATGCGG AGCATATACG CCCGGAGCCG CGGCGATCCT 2161 GCAAGCTCCG GATGCCTCCG CTCGAAGTAG CGCGTCTGCT GCTCCATACA AGCCAACCAC 2221 GGCCTCCAGA AGAAGATGTT GGCGACCTCG TATTGGGAAT CCCCGAACAT CGCCTCGCTC 2281 CAGTCAATGA CCGCTGTTAT GCGGCCATTG TCCGTCAGGA CATTGTTGGA GCCGAAATCC 2341 GCGTGCACGA GGTGCCGGAC TTCGGGGCAG TCCTCGGCCC AAAGCATCAG CTCATCGAGA 2401 GCCTGCGCGA CGGACGCACT GACGGTGTCG TCCATCACAG TTTGCCAGTG ATACACATGG 2461 GGATCAGCAA TCGCGCATAT GAAATCACGC CATGTAGTGT ATTGACCGAT TCCTTGCGGT 2521 CCGAATGGGC CGAACCCGCT CGTCTGGCTA AGATCGGCCG CAGCGATCGC ATCCATGGCC 2581 TCCGCGACCG GCTGCAGAAC AGCGGGCAGT TCGGTTTCAG GCAGGTCTTG CAACGTGACA 2641 CCCTGTGCAC GGCGGGAGAT GCAATAGGTC AGGCTCTCGC TGAATTCCCC AATGTCAAGC 2701 ACTTCCGGAA TCGGGAGCGC GGCCGATGCA AAGTGCCGAT AAACATAACG ATCTTTGTAG 2761 AAACCATCGG CGCAGCTATT TACCCGCAGG ACATATCCAC GCCCTCCTAC ATCGAAGCTG 2821 AAAGCACGAG ATTCTTCGCC CTCCGAGAGC TGCATCAGGT CGGAGACGCT GTCGAACTTT 2881 TCGATCAGAA ACTTCTCGAC AGACGTCGCG GTGAGTTCAG GCTTTTTCAT CATTTACCAA 2941 CAGTAagatc tGTAATGTCA GTGCGAGCGA TCAAGAATGT AACACAAAGG AATaCCAAGT 3001 TTGTGAATTT TATTTTTTGG TGTAATTGAA GtCTTCAAGT TGAAAAAAAT GGGGAAAAGG 3061 AAAAAAAACT CGAAGGCGCG TGCAGCggcc ggccGACGAG AACgTGTacc ggtATAACTT 3121 CGTATAATGT ATGCTATACG AAGTTATAAG CTTATCGTCG ACtATGCGTA ATCGGGCACA 3181 TCGTACGGGT ATGCGTAGTC TGGCACGTCG TATGGGTACG CGTAATCAGG CACATCGTAA 3241 GGGTACTCGA gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg 3301 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 3361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 3421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 3481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 3541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 3601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 3661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 3721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 3781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 3841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 3901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 3961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 4021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 4081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 4141 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 4201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 4261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 4321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 4381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 4441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 4501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 4561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 4621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 4681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 4741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 4801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 4861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 4921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 4981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 5041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 5101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 5161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 5221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 5281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 5341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 5401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 5461 gtgccac //