LOCUS pCR_Blunt_II_TOPO 4001 bp ds-DNA circular 06-AUG-2014 DEFINITION . ACCESSION VERSION SOURCE . ORGANISM . COMMENT Insert is blunt cloned at beginning or end of sequence COMMENT ApEinfo:methylated:1 FEATURES Location/Qualifiers CDS 822..1031 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=ccdB /ApEinfo_fwdcolor=yellow /ApEinfo_revcolor=yellow primer_bind complement(578..595) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=M13-fwd /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green primer_bind complement(552..579) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=T7 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green enhancer complement(192..199) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=Putative Enhancer /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green rep_origin 2820..3502 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=ColE1 origin /ApEinfo_fwdcolor=gray50 /ApEinfo_revcolor=gray50 misc_RNA 482..482 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=gRNA /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green 3'UTR 1..66 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=tbb-2 3'UTR /ApEinfo_fwdcolor=#cc3333 /ApEinfo_revcolor=#cc3333 CDS 1386..2177 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=Kan/neoR /ApEinfo_fwdcolor=yellow /ApEinfo_revcolor=yellow primer_bind 3870..3890 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=M13-rev /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green primer_bind 3896..3921 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=SP6 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green misc_binding 3842..3864 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=LacO /ApEinfo_fwdcolor=cornflower blue /ApEinfo_revcolor=cornflower blue misc_feature 92..131 /translation="MGKIQKVGRWVPHELNERQMERRKNTCEILLSRYKRKSFLHRIV TGDEKWIFFVNPKRKKSYVDPGQPATSTARPNRFGKKTMLCVWWDQSGVIYYELLKPG ETVNTARYQQQLINLNRALQRKRPEYQKRQHRVIFLHDNAPSHTARAVRDTLETLNWE VLPHAAYSPDLAPSDYHLFASMGHALAEQRFDSYESVKKWLDEWFAAKDDEFYWRGIH KLPERWEKCVASDGKYFE*" /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=lacZ_a /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink promoter 132..481 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=U6 Promoter /ApEinfo_fwdcolor=#33cc00 /ApEinfo_revcolor=#33cc00 misc_recomb complement(67..87) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=attB3 /ApEinfo_fwdcolor=#dda0dd /ApEinfo_revcolor=#dda0dd misc_feature complement(74..80) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=Gateway Recomb3 /ApEinfo_fwdcolor=#ff0000 /ApEinfo_revcolor=#ff0000 misc_feature complement(104..126) /translation="MGKIQKVGRWVPHELNERQMERRKNTCEILLSRYKRKSFLHRIV TGDEKWIFFVNPKRKKSYVDPGQPATSTARPNRFGKKTMLCVWWDQSGVIYYELLKPG ETVNTARYQQQLINLNRALQRKRPEYQKRQHRVIFLHDNAPSHTARAVRDTLETLNWE VLPHAAYSPDLAPSDYHLFASMGHALAEQRFDSYESVKKWLDEWFAAKDDEFYWRGIH KLPERWEKCVASDGKYFE*" /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=M13_pUC_fwd_primer /ApEinfo_fwdcolor=pink /ApEinfo_revcolor=pink primer_bind complement(95..112) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=M13-fwd(1) /ApEinfo_label=M13-fwd /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green primer_bind complement(47..66) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=tbb2T R #68 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green primer_bind complement(112..131) /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 /label=Gateway Promoter L #73 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green ORIGIN 1 attgtgttcg ttgagtgacc caacaaaaag aggaaacttt attgtgccgc caagaaaaaa 61 gtctcaCAAC TTTattatac atagttgata attcactggc cgtcgtttta caacgtcgtg 121 actgggaaaa ccaaaaaaaa ctagcaataa aggaataaaa aactgtacac cttaaaggcg 181 cacactctgt tttgcaaatt ttatttttag ttgtgaattt tctgctgaga cctgaaaata 241 gcaactttag tactactata atttgtcaac cttttcaaaa aaagcatgca atttttgaga 301 aactcttata aaagctatta ttaaaaaaac accttttttc caaaattatt ccacaaaaaa 361 tatgttatga aatgcctaca ccctctcaca cacactcttt atactactct gtcaaactca 421 cgagatgtct gccgcctctt gtgttgcccc tatataaaca cctcctattg cgagatgtct 481 tGAAGGGCGA ATTCTGCAGA TATCCATCAC ACTGGCGGCC GCTCGAGCAT GCATCTAGAG 541 GGCCCAATTC GCCCTATAGT GAGTCGTATT ACAATTCACT GGCCGTCGTT TTACAACGTC 601 GTGACTGGGA AAACCCTGGC GTTACCCAAC TTAATCGCCT TGCAGCACAT CCCCCTTTCG 661 CCAGCTGGCG TAATAGCGAA GAGGCCCGCA CCGATCGCCC TTCCCAACAG TTGCGCAGCC 721 TATACGTACG GCAGTTTAAG GTTTACACCT ATAAAAGAGA GAGCCGTTAT CGTCTGTTTG 781 TGGATGTACA GAGTGATATT ATTGACACGC CGGGGCGACG GATGGTGATC CCCCTGGCCA 841 GTGCACGTCT GCTGTCAGAT AAAGTCTCCC GTGAACTTTA CCCGGTGGTG CATATCGGGG 901 ATGAAAGCTG GCGCATGATG ACCACCGATA TGGCCAGTGT GCCGGTCTCC GTTATCGGGG 961 AAGAAGTGGC TGATCTCAGC CACCGCGAAA ATGACATCAA AAACGCCATT AACCTGATGT 1021 TCTGGGGAAT ATAAATGTCA GGCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT 1081 TCACGTAGAA AGCCAGTCCG CAGAAACGGT GCTGACCCCG GATGAATGTC AGCTACTGGG 1141 CTATCTGGAC AAGGGAAAAC GCAAGCGCAA AGAGAAAGCA GGTAGCTTGC AGTGGGCTTA 1201 CATGGCGATA GCTAGACTGG GCGGTTTTAT GGACAGCAAG CGAACCGGAA TTGCCAGCTG 1261 GGGCGCCCTC TGGTAAGGTT GGGAAGCCCT GCAAAGTAAA CTGGATGGCT TTCTCGCCGC 1321 CAAGGATCTG ATGGCGCAGG GGATCAAGCT CTGATCAAGA GACAGGATGA GGATCGTTTC 1381 GCATGATTGA ACAAGATGGA TTGCACGCAG GTTCTCCGGC CGCTTGGGTG GAGAGGCTAT 1441 TCGGCTATGA CTGGGCACAA CAGACAATCG GCTGCTCTGA TGCCGCCGTG TTCCGGCTGT 1501 CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA AGACCGACCT GTCCGGTGCC CTGAATGAAC 1561 TGCAAGACGA GGCAGCGCGG CTATCGTGGC TGGCCACGAC GGGCGTTCCT TGCGCAGCTG 1621 TGCTCGACGT TGTCACTGAA GCGGGAAGGG ACTGGCTGCT ATTGGGCGAA GTGCCGGGGC 1681 AGGATCTCCT GTCATCTCAC CTTGCTCCTG CCGAGAAAGT ATCCATCATG GCTGATGCAA 1741 TGCGGCGGCT GCATACGCTT GATCCGGCTA CCTGCCCATT CGACCACCAA GCGAAACATC 1801 GCATCGAGCG AGCACGTACT CGGATGGAAG CCGGTCTTGT CGATCAGGAT GATCTGGACG 1861 AAGAGCATCA GGGGCTCGCG CCAGCCGAAC TGTTCGCCAG GCTCAAGGCG AGCATGCCCG 1921 ACGGCGAGGA TCTCGTCGTG ACCCATGGCG ATGCCTGCTT GCCGAATATC ATGGTGGAAA 1981 ATGGCCGCTT TTCTGGATTC ATCGACTGTG GCCGGCTGGG TGTGGCGGAC CGCTATCAGG 2041 ACATAGCGTT GGCTACCCGT GATATTGCTG AAGAGCTTGG CGGCGAATGG GCTGACCGCT 2101 TCCTCGTGCT TTACGGTATC GCCGCTCCCG ATTCGCAGCG CATCGCCTTC TATCGCCTTC 2161 TTGACGAGTT CTTCTGAATT ATTAACGCTT ACAATTTCCT GATGCGGTAT TTTCTCCTTA 2221 CGCATCTGTG CGGTATTTCA CACCGCATAC AGGTGGCACT TTTCGGGGAA ATGTGCGCGG 2281 AACCCCTATT TGTTTATTTT TCTAAATACA TTCAAATATG TATCCGCTCA TGAGACAATA 2341 ACCCTGATAA ATGCTTCAAT AATAGCACGT GAGGAGGGCC ACCATGGCCA AGTTGACCAG 2401 TGCCGTTCCG GTGCTCACCG CGCGCGACGT CGCCGGAGCG GTCGAGTTCT GGACCGACCG 2461 GCTCGGGTTC TCCCGGGACT TCGTGGAGGA CGACTTCGCC GGTGTGGTCC GGGACGACGT 2521 GACCCTGTTC ATCAGCGCGG TCCAGGACCA GGTGGTGCCG GACAACACCC TGGCCTGGGT 2581 GTGGGTGCGC GGCCTGGACG AGCTGTACGC CGAGTGGTCG GAGGTCGTGT CCACGAACTT 2641 CCGGGACGCC TCCGGGCCGG CCATGACCGA GATCGGCGAG CAGCCGTGGG GGCGGGAGTT 2701 CGCCCTGCGC GACCCGGCCG GCAACTGCGT GCACTTCGTG GCCGAGGAGC AGGACTGACA 2761 CGTGCTAAAA CTTCATTTTT AATTTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT 2821 CATGACCAAA ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA 2881 GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT TGCAAACAAA 2941 AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA GAGCTACCAA CTCTTTTTCC 3001 GAAGGTAACT GGCTTCAGCA GAGCGCAGAT ACCAAATACT GTCCTTCTAG TGTAGCCGTA 3061 GTTAGGCCAC CACTTCAAGA ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT 3121 GTTACCAGTG GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG 3181 ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA CACAGCCCAG 3241 CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG CGTGAGCTAT GAGAAAGCGC 3301 CACGCTTCCC GAAGGGAGAA AGGCGGACAG GTATCCGGTA AGCGGCAGGG TCGGAACAGG 3361 AGAGCGCACG AGGGAGCTTC CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT 3421 TCGCCACCTC TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG 3481 GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGGC TTTTGCTGGC CTTTTGCTCA 3541 CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC CGTATTACCG CCTTTGAGTG 3601 AGCTGATACC GCTCGCCGCA GCCGAACGAC CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC 3661 GGAAGAGCGC CCAATACGCA AACCGCCTCT CCCCGCGCGT TGGCCGATTC ATTAATGCAG 3721 CTGGCACGAC AGGTTTCCCG ACTGGAAAGC GGGCAGTGAG CGCAACGCAA TTAATGTGAG 3781 TTAGCTCACT CATTAGGCAC CCCAGGCTTT ACACTTTATG CTTCCGGCTC GTATGTTGTG 3841 TGGAATTGTG AGCGGATAAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGCCAA 3901 GCTATTTAGG TGACACTATA GAATACTCAA GCTATGCATC AAGCTTGGTA CCGAGCTCGG 3961 ATCCACTAGT AACGGCCGCC AGTGTGCTGG AATTCGCCCT T //