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   
//
