LOCUS pGTag_eGFP_caax_SV40 4164 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:1 FEATURES Location/Qualifiers misc_feature complement(617..622) /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 1031..1096 /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" primer_bind 536..551 /locus_tag="M13F" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" terminator 267..294 /locus_tag="T2" /ApEinfo_fwdcolor="#bbbbbb" /ApEinfo_revcolor="#bbbbbb" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 745..1020 /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 590..612 /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(609..612) /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 1021..1026 /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" terminator 426..469 /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 1031..1034 /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" promoter 623..744 /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" misc_feature 1097..1813 /locus_tag="eGFP" /ApEinfo_fwdcolor="#00ff00" /ApEinfo_revcolor="green" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_recomb 2111..2206 /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 2111..2125 /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" misc_feature complement(2352..2374) /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" polyA_signal 1914..2109 /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" misc_structure 1814..1873 /locus_tag="CAAX box from Harvey ras" /ApEinfo_fwdcolor="#ff0000" /ApEinfo_revcolor="#ff0000" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" rep_origin 3487..4160 /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" CDS 2557..3366 /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(2424..2444) /locus_tag="M13R" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature complement(2238..2244) /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 2352..2354 /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" promoter complement(2401..2420) /locus_tag="T7" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" intron 2253..2341 /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 2344..2350 /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_feature complement(2234..2236) /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" ORIGIN 1 TTTCCTGCGT TATCCCCTGA TTCTGTGGAT AACCGTATTA CCGCCTTTGA GTGAGCTGAT 61 ACCGCTCGCC GCAGCCGAAC GACCGAGCGC AGCGAGTCAG TGAGCGAGGA AGCGGAtGtG 121 CGCCCAATAC GCAAACCGCC TCTCCCCGCG CGTTGGCCGA TTCATTAATG CAGCTGGCAC 181 GACAGGTTTC CCGACTGGAA AGCGGGCAGT GAGCGCAACG CAATTAATAC GCGTACCGCT 241 AGCCAGGAAG AGTTTGTAGA AACGCAAAAA GGCCATCCGT CAGGATGGCC TTCTGCTTAG 301 TTTGATGCCT GGCAGTTTAT GGCGGGCGTC CTGCCCGCCA CCCTCCGGGC CGTTGCTTCA 361 CAACGTTCAA ATCCGCTCCC GGCGGATTTG TCCTACTCAG GAGAGCGTTC ACCGACAAAC 421 AACAGATAAA ACGAAAGGCC CAGTCTTCCG ACTGAGCCTT TCGTTTTATT TGATGCCTGG 481 CAGTTCCCTA CTCTCGCGTT AACGCTAGCA TGGATGTTTT CCCAGTCACG ACGTTGTAAA 541 ACGACGGCCA GTCTTAAGCT CGGGCCCTGC AGCTCTAGAG CTCGAATTCG GGAGGCGTTC 601 GGGCCACAGC GGacacgcag gtgcgcaacg caattaatgt gagttagctc actcattagg 661 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat 721 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc 781 actaaaggga acaaaagctg gaggacctcg agggggggcc cggtacccaa ttcgccctat 841 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac 901 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat 961 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg 1021 acctgccaca GGATCCGGAG CCACGAACTT CTCTCTGTTA AAGCAAGCAG GAGACGTGGA 1081 AGAAAACCCC GGTCCTatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 1141 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 1201 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 1261 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 1321 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 1381 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 1441 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 1501 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 1561 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 1621 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 1681 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 1741 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 1801 cgagctgtac aagAAGCTGA ACCCTCCTGA TGAGAGTGGC CCCGGCTGCA TGAGCTGCAA 1861 GTGTGTGCTC TCCTGAGGCG CGCCTCTAGA ACTATAGTGA GTCGTATTAC GTAGATCCAG 1921 ACATGATAAG ATACATTGAT GAGTTTGGAC AAACCACAAC TAGAATGCAG TGAAAAAAAT 1981 GCTTTATTTG TGAAATTTGT GATGCTATTG CTTTATTTGT AACCATTATA AGCTGCAATA 2041 AACAAGTTAA CAACAACAAT TGCATTCATT TTATGTTTCA GGTTCAGGGG GAGGTGTGGG 2101 AGGTTTTTTC CAACTTTATT ATACAAAGTT GGCATTATAA AAAAGCATTG CTTATCAATT 2161 TGTTGCAACG AACAGGTCAC TATCAGTCAA AATAAAATCA TTATTTGGAG CTCCATGGcg 2221 ttgtctagca aggaagtgaa gagctatccg gatatatatg tatgttatca taactgtact 2281 ggctatgtca ggtggtaatg actgtaacgt tacgttactc gttgtaggca cgacattgaa 2341 tatgctcttc tCCGCTGTGG CCCGAACGCC TCCCgctagc gatgggatct CGGCCGCGAT 2401 ATCCCCTATA GTGAGTCGTA TTACATGGTC ATAGCTGTTT CCTGGCAGCT CTGGCCCGTG 2461 TCTCAAAATC TCTGATGTTA CATTGCACAA GATAAAAATA TATCATCATG AACAATAAAA 2521 CTGTCTGCTT ACATAAACAG TAATACAAGG GGTGTTATGA GCCATATTCA ACGGGAAACG 2581 TCGAGGCCGC GATTAAATTC CAACATGGAT GCTGATTTAT ATGGGTATAA ATGGGCTCGC 2641 GATAATGTCG GGCAATCAGG TGCGACAATC TATCGCTTGT ATGGGAAGCC CGATGCGCCA 2701 GAGTTGTTTC TGAAACATGG CAAAGGTAGC GTTGCCAATG ATGTTACAGA TGAGATGGTC 2761 AGACTAAACT GGCTGACGGA ATTTATGCCT CTTCCGACCA TCAAGCATTT TATCCGTACT 2821 CCTGATGATG CATGGTTACT CACCACTGCG ATCCCCGGAA AAACAGCATT CCAGGTATTA 2881 GAAGAATATC CTGATTCAGG TGAAAATATT GTTGATGCGC TGGCAGTGTT CCTGCGCCGG 2941 TTGCATTCGA TTCCTGTTTG TAATTGTCCT TTTAACAGCG ATCGCGTATT TCGTCTCGCT 3001 CAGGCGCAAT CACGAATGAA TAACGGTTTG GTTGATGCGA GTGATTTTGA TGACGAGCGT 3061 AATGGCTGGC CTGTTGAACA AGTCTGGAAA GAAATGCATA AACTTTTGCC ATTCTCACCG 3121 GATTCAGTCG TCACTCATGG TGATTTCTCA CTTGATAACC TTATTTTTGA CGAGGGGAAA 3181 TTAATAGGTT GTATTGATGT TGGACGAGTC GGAATCGCAG ACCGATACCA GGATCTTGCC 3241 ATCCTATGGA ACTGCCTCGG TGAGTTTTCT CCTTCATTAC AGAAACGGCT TTTTCAAAAA 3301 TATGGTATTG ATAATCCTGA TATGAATAAA TTGCAGTTTC ATTTGATGCT CGATGAGTTT 3361 TTCTAATCAG AATTGGTTAA TTGGTTGTAA CACTGGCAGA GCATTACGCT GACTTGACGG 3421 GACGGCGCAA GCTCATGACC AAAATCCCTT AACGTGAGTT ACGCGTCGTT CCACTGAGCG 3481 TCAGACCCCG TAGAAAAGAT CAAAGGATCT TCTTGAGATC CTTTTTTTCT GCGCGTAATC 3541 TGCTGCTTGC AAACAAAAAA ACCACCGCTA CCAGCGGTGG TTTGTTTGCC GGATCAAGAG 3601 CTACCAACTC TTTTTCCGAA GGTAACTGGC TTCAGCAGAG CGCAGATACC AAATACTGTC 3661 CTTCTAGTGT AGCCGTAGTT AGGCCACCAC TTCAAGAACT CTGTAGCACC GCCTACATAC 3721 CTCGCTCTGC TAATCCTGTT ACCAGTGGCT GCTGCCAGTG GCGATAAGTC GTGTCTTACC 3781 GGGTTGGACT CAAGACGATA GTTACCGGAT AAGGCGCAGC GGTCGGGCTG AACGGGGGGT 3841 TCGTGCACAC AGCCCAGCTT GGAGCGAACG ACCTACACCG AACTGAGATA CCTACAGCGT 3901 GAGCATTGAG AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG CGGACAGGTA TCCGGTAAGC 3961 GGCAGGGTCG GAACAGGAGA GCGCACGAGG GAGCTTCCAG GGGGAAACGC CTGGTATCTT 4021 TATAGTCCTG TCGGGTTTCG CCACCTCTGA CTTGAGCGTC GATTTTTGTG ATGCTCGTCA 4081 GGGGGGCGGA GCCTATGGAA AAACGCCAGC AACGCGGCCT TTTTACGGTT CCTGGCCTTT 4141 TGCTGGCCTT TTGCTCACAT GTTC //