LOCUS pGTag_TagRFP_SV40 4098 bp DNA circular 16-JUL-2018 DEFINITION synthetic circular DNA, gene-targeting cassette ACCESSION . VERSION 2.0 KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 4786) AUTHORS Wesley A Wierson, Jordan M Welker, Maira P Almeida, Carla M Mann, Dennis A Webster, Trevor J Weiss, Macy K Vollbrecht, Zhitao Ming, Alec Wehmeier, Christopher S Mikelson, Jeffrey A Haltom, Kristen M Kwan, Chi-Bin Chien, Darius Balciunas, Stephen C Ekker, Karl J Clark, Beau R Webber, Branden Moriarity, Staci L Solin, Daniel F Carlson, Drena L Dobbs, Maura McGrail, Jeffrey J Essner TITLE Efficient targeted integration directed by short homology JOURNAL . COMMENT COMMENT ApEinfo:methylated:0 FEATURES Location/Qualifiers rep_origin 3422..4095 /locus_tag="pUC ori" /ApEinfo_fwdcolor="#999999" /ApEinfo_revcolor="#999999" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 1032..1097 /locus_tag="PTV1-2A peptide" /ApEinfo_fwdcolor="#004fff" /ApEinfo_revcolor="cyan" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" CDS 2492..3301 /locus_tag="kanR" /ApEinfo_fwdcolor="#ff7f3f" /ApEinfo_revcolor="#ff7f3f" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" primer_bind complement(2359..2379) /locus_tag="M13R" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" primer_bind 537..552 /locus_tag="M13F" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" promoter complement(2336..2355) /locus_tag="T7" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" terminator 268..295 /locus_tag="T2" /ApEinfo_fwdcolor="#bbbbbb" /ApEinfo_revcolor="#bbbbbb" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" terminator 427..470 /locus_tag="T1" /ApEinfo_fwdcolor="#bbbbbb" /ApEinfo_revcolor="#bbbbbb" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature complement(2287..2309) /locus_tag="3' Universal Guide (w/PAM)" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature complement(618..623) /locus_tag="BfuAI site 1" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature complement(2173..2179) /locus_tag="BspQI site 1" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 746..1021 /vntifkey="21" /locus_tag="lacZ'" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="#1dda7b" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 591..613 /locus_tag="5' universal guide (w/PAM)" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature complement(2169..2171) /locus_tag="BspQI site 1 overhang" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" promoter 624..745 /vntifkey="30" /locus_tag="lacZ\promoter" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" intron 2188..2276 /note="Carp beta-actin intron 1 filler" /locus_tag="Carp beta-actin intron 1 filler" /ApEinfo_fwdcolor="pink" /ApEinfo_revcolor="pink" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature complement(610..613) /locus_tag="BfuAI site 1 overhang" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 2287..2289 /locus_tag="BspQI site 2 overhang" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 1022..1027 /locus_tag="BfuAI site 2" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 2279..2285 /locus_tag="BspQI site 2" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_recomb 2046..2141 /locus_tag="attL3" /ApEinfo_fwdcolor="#5370ff" /ApEinfo_revcolor="#5370ff" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 2046..2060 /locus_tag="att3_shared" /ApEinfo_fwdcolor="#4259cc" /ApEinfo_revcolor="#4259cc" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" polyA_signal 1849..2044 /locus_tag="SV40 late polyA (from pCS2+)" /ApEinfo_fwdcolor="#ffff7f" /ApEinfo_revcolor="#ffff7f" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" CDS 1098..1808 /locus_tag="TagRFP" /ApEinfo_fwdcolor="#cc0000" /ApEinfo_revcolor="#cc0000" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 1032..1035 /locus_tag="BfuAI site 2 overhang" /ApEinfo_fwdcolor="cyan" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" ORIGIN 1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA 61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAtGt 121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA 181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC 241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA 301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC 361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA 421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG 481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA 541 AACGACGGCC AGTCTTAAGC TCGGGCCCTG CAGCTCTAGA GCTCGAATTC GGGAGGCGTT 601 CGGGCCACAG CGGacacgca ggtgcgcaac gcaattaatg tgagttagct cactcattag 661 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga 721 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct 781 cactaaaggg aacaaaagct ggaggacctc gagggggggc ccggtaccca attcgcccta 841 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa 901 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 961 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 1021 gacctgccac aGGATCCGGA GCCACGAACT TCTCTCTGTT AAAGCAAGCA GGAGACGTGG 1081 AAGAAAACCC CGGTCCTATG GTGTCTAAGG GCGAgGAaCT GATTAAGGAG AACATGCACA 1141 TGAAGCTGTA CATGGAGGGC ACCGTGAACA ACCACCACTT CAAGTGCACA TCCGAGGGCG 1201 AAGGCAAGCC CTACGAGGGC ACCCAGACCA TGAGAATCAA GGTGGTCGAG GGCGGCCCTC 1261 TCCCCTTCGC CTTCGACATC CTGGCTACCA GCTTCATGTA CGGCAGCAGA ACCTTCATCA 1321 ACCACACCCA GGGCATCCCC GACTTCTTTA AGCAGTCCTT CCCTGAGGGC TTCACATGGG 1381 AGAGAGTCAC CACATACGAA GACGGGGGCG TGCTGACCGC TACCCAGGAC ACCAGCCTCC 1441 AGGACGGCTG CCTCATCTAC AACGTCAAGA TCAGAGGGGT GAACTTCCCA TCCAACGGCC 1501 CTGTGATGCA GAAGAAAACA CTCGGCTGGG AGGCCAACAC CGAGATGCTG TACCCCGCTG 1561 ACGGCGGCCT GGAAGGCAGA AGCGACATGG CCCTGAAGCT CGTGGGCGGG GGCCACCTGA 1621 TCTGCAACTT CAAGACCACA TACAGATCCA AGAAACCCGC TAAGAACCTC AAGATGCCCG 1681 GCGTCTACTA TGTGGACCAC AGACTGGAAA GAATCAAGGA GGCCGACAAA GAGACCTACG 1741 TCGAGCAGCA CGAGGTGGCT GTGGCCAGAT ACTGCGACCT CCCTAGCAAA CTGGGGCACA 1801 AACTTAATTG AGGCGCGCCT CTAGAACTAT AGTGAGTCGT ATTACGTAGA TCCAGACATG 1861 ATAAGATACA TTGATGAGTT TGGACAAACC ACAACTAGAA TGCAGTGAAA AAAATGCTTT 1921 ATTTGTGAAA TTTGTGATGC TATTGCTTTA TTTGTAACCA TTATAAGCTG CAATAAACAA 1981 GTTAACAACA ACAATTGCAT TCATTTTATG TTTCAGGTTC AGGGGGAGGT GTGGGAGGTT 2041 TTTTCCAACT TTATTATACA AAGTTGGCAT TATAAAAAAG CATTGCTTAT CAATTTGTTG 2101 CAACGAACAG GTCACTATCA GTCAAAATAA AATCATTATT TGGAGCTCCA TGGcgttgtc 2161 tagcaaggaa gtgaagagct atccggatat atatgtatgt tatcataact gtactggcta 2221 tgtcaggtgg taatgactgt aacgttacgt tactcgttgt aggcacgaca ttgaatatgc 2281 tcttctCCGC TGTGGCCCGA ACGCCTCCCg ctagcgatgg gatctCGGCC GCGATATCCC 2341 CTATAGTGAG TCGTATTACA TGGTCATAGC TGTTTCCTGG CAGCTCTGGC CCGTGTCTCA 2401 AAATCTCTGA TGTTACATTG CACAAGATAA AAATATATCA TCATGAACAA TAAAACTGTC 2461 TGCTTACATA AACAGTAATA CAAGGGGTGT TATGAGCCAT ATTCAACGGG AAACGTCGAG 2521 GCCGCGATTA AATTCCAACA TGGATGCTGA TTTATATGGG TATAAATGGG CTCGCGATAA 2581 TGTCGGGCAA TCAGGTGCGA CAATCTATCG CTTGTATGGG AAGCCCGATG CGCCAGAGTT 2641 GTTTCTGAAA CATGGCAAAG GTAGCGTTGC CAATGATGTT ACAGATGAGA TGGTCAGACT 2701 AAACTGGCTG ACGGAATTTA TGCCTCTTCC GACCATCAAG CATTTTATCC GTACTCCTGA 2761 TGATGCATGG TTACTCACCA CTGCGATCCC CGGAAAAACA GCATTCCAGG TATTAGAAGA 2821 ATATCCTGAT TCAGGTGAAA ATATTGTTGA TGCGCTGGCA GTGTTCCTGC GCCGGTTGCA 2881 TTCGATTCCT GTTTGTAATT GTCCTTTTAA CAGCGATCGC GTATTTCGTC TCGCTCAGGC 2941 GCAATCACGA ATGAATAACG GTTTGGTTGA TGCGAGTGAT TTTGATGACG AGCGTAATGG 3001 CTGGCCTGTT GAACAAGTCT GGAAAGAAAT GCATAAACTT TTGCCATTCT CACCGGATTC 3061 AGTCGTCACT CATGGTGATT TCTCACTTGA TAACCTTATT TTTGACGAGG GGAAATTAAT 3121 AGGTTGTATT GATGTTGGAC GAGTCGGAAT CGCAGACCGA TACCAGGATC TTGCCATCCT 3181 ATGGAACTGC CTCGGTGAGT TTTCTCCTTC ATTACAGAAA CGGCTTTTTC AAAAATATGG 3241 TATTGATAAT CCTGATATGA ATAAATTGCA GTTTCATTTG ATGCTCGATG AGTTTTTCTA 3301 ATCAGAATTG GTTAATTGGT TGTAACACTG GCAGAGCATT ACGCTGACTT GACGGGACGG 3361 CGCAAGCTCA TGACCAAAAT CCCTTAACGT GAGTTACGCG TCGTTCCACT GAGCGTCAGA 3421 CCCCGTAGAA AAGATCAAAG GATCTTCTTG AGATCCTTTT TTTCTGCGCG TAATCTGCTG 3481 CTTGCAAACA AAAAAACCAC CGCTACCAGC GGTGGTTTGT TTGCCGGATC AAGAGCTACC 3541 AACTCTTTTT CCGAAGGTAA CTGGCTTCAG CAGAGCGCAG ATACCAAATA CTGTCCTTCT 3601 AGTGTAGCCG TAGTTAGGCC ACCACTTCAA GAACTCTGTA GCACCGCCTA CATACCTCGC 3661 TCTGCTAATC CTGTTACCAG TGGCTGCTGC CAGTGGCGAT AAGTCGTGTC TTACCGGGTT 3721 GGACTCAAGA CGATAGTTAC CGGATAAGGC GCAGCGGTCG GGCTGAACGG GGGGTTCGTG 3781 CACACAGCCC AGCTTGGAGC GAACGACCTA CACCGAACTG AGATACCTAC AGCGTGAGCA 3841 TTGAGAAAGC GCCACGCTTC CCGAAGGGAG AAAGGCGGAC AGGTATCCGG TAAGCGGCAG 3901 GGTCGGAACA GGAGAGCGCA CGAGGGAGCT TCCAGGGGGA AACGCCTGGT ATCTTTATAG 3961 TCCTGTCGGG TTTCGCCACC TCTGACTTGA GCGTCGATTT TTGTGATGCT CGTCAGGGGG 4021 GCGGAGCCTA TGGAAAAACG CCAGCAACGC GGCCTTTTTA CGGTTCCTGG CCTTTTGCTG 4081 GCCTTTTGCT CACATGTT //