LOCUS pGTag_eGFP_SV40 4110 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 misc_recomb 2058..2153 /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 2058..2072 /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 1861..2056 /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_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" rep_origin 3434..4107 /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 2504..3313 /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(2371..2391) /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(2348..2367) /locus_tag="T7" /ApEinfo_fwdcolor="#777777" /ApEinfo_revcolor="#777777" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" misc_feature 1098..1817 /locus_tag="eGFP" /ApEinfo_fwdcolor="#00ff00" /ApEinfo_revcolor="green" /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 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" misc_feature complement(2299..2321) /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(2185..2191) /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(2181..2183) /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 2200..2288 /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 2299..2301 /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 2291..2297 /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" 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 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 1141 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 1201 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 1261 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct 1321 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 1381 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 1441 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 1501 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg 1561 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg 1621 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 1681 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga 1741 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 1801 acgagctgta caagtaactc gagGGCGCGC CTCTAGAACT ATAGTGAGTC GTATTACGTA 1861 GATCCAGACA TGATAAGATA CATTGATGAG TTTGGACAAA CCACAACTAG AATGCAGTGA 1921 AAAAAATGCT TTATTTGTGA AATTTGTGAT GCTATTGCTT TATTTGTAAC CATTATAAGC 1981 TGCAATAAAC AAGTTAACAA CAACAATTGC ATTCATTTTA TGTTTCAGGT TCAGGGGGAG 2041 GTGTGGGAGG TTTTTTCCAA CTTTATTATA CAAAGTTGGC ATTATAAAAA AGCATTGCTT 2101 ATCAATTTGT TGCAACGAAC AGGTCACTAT CAGTCAAAAT AAAATCATTA TTTGGAGCTC 2161 CATGGcgttg tctagcaagg aagtgaagag ctatccggat atatatgtat gttatcataa 2221 ctgtactggc tatgtcaggt ggtaatgact gtaacgttac gttactcgtt gtaggcacga 2281 cattgaatat gctcttctCC GCTGTGGCCC GAACGCCTCC Cgctagcgat gggatctCGG 2341 CCGCGATATC CCCTATAGTG AGTCGTATTA CATGGTCATA GCTGTTTCCT GGCAGCTCTG 2401 GCCCGTGTCT CAAAATCTCT GATGTTACAT TGCACAAGAT AAAAATATAT CATCATGAAC 2461 AATAAAACTG TCTGCTTACA TAAACAGTAA TACAAGGGGT GTTATGAGCC ATATTCAACG 2521 GGAAACGTCG AGGCCGCGAT TAAATTCCAA CATGGATGCT GATTTATATG GGTATAAATG 2581 GGCTCGCGAT AATGTCGGGC AATCAGGTGC GACAATCTAT CGCTTGTATG GGAAGCCCGA 2641 TGCGCCAGAG TTGTTTCTGA AACATGGCAA AGGTAGCGTT GCCAATGATG TTACAGATGA 2701 GATGGTCAGA CTAAACTGGC TGACGGAATT TATGCCTCTT CCGACCATCA AGCATTTTAT 2761 CCGTACTCCT GATGATGCAT GGTTACTCAC CACTGCGATC CCCGGAAAAA CAGCATTCCA 2821 GGTATTAGAA GAATATCCTG ATTCAGGTGA AAATATTGTT GATGCGCTGG CAGTGTTCCT 2881 GCGCCGGTTG CATTCGATTC CTGTTTGTAA TTGTCCTTTT AACAGCGATC GCGTATTTCG 2941 TCTCGCTCAG GCGCAATCAC GAATGAATAA CGGTTTGGTT GATGCGAGTG ATTTTGATGA 3001 CGAGCGTAAT GGCTGGCCTG TTGAACAAGT CTGGAAAGAA ATGCATAAAC TTTTGCCATT 3061 CTCACCGGAT TCAGTCGTCA CTCATGGTGA TTTCTCACTT GATAACCTTA TTTTTGACGA 3121 GGGGAAATTA ATAGGTTGTA TTGATGTTGG ACGAGTCGGA ATCGCAGACC GATACCAGGA 3181 TCTTGCCATC CTATGGAACT GCCTCGGTGA GTTTTCTCCT TCATTACAGA AACGGCTTTT 3241 TCAAAAATAT GGTATTGATA ATCCTGATAT GAATAAATTG CAGTTTCATT TGATGCTCGA 3301 TGAGTTTTTC TAATCAGAAT TGGTTAATTG GTTGTAACAC TGGCAGAGCA TTACGCTGAC 3361 TTGACGGGAC GGCGCAAGCT CATGACCAAA ATCCCTTAAC GTGAGTTACG CGTCGTTCCA 3421 CTGAGCGTCA GACCCCGTAG AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG 3481 CGTAATCTGC TGCTTGCAAA CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA 3541 TCAAGAGCTA CCAACTCTTT TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA 3601 TACTGTCCTT CTAGTGTAGC CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC 3661 TACATACCTC GCTCTGCTAA TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG 3721 TCTTACCGGG TTGGACTCAA GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC 3781 GGGGGGTTCG TGCACACAGC CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT 3841 ACAGCGTGAG CATTGAGAAA GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC 3901 GGTAAGCGGC AGGGTCGGAA CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG 3961 GTATCTTTAT AGTCCTGTCG GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG 4021 CTCGTCAGGG GGGCGGAGCC TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT 4081 GGCCTTTTGC TGGCCTTTTG CTCACATGTT //