LOCUS       pGTag_TagRFP_caax_SV40        4158 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     2106..2201
                     /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    2106..2120
                     /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(2347..2369)
                     /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"
     polyA_signal    1909..2104
                     /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"
     misc_structure  1809..1868
                     /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      3482..4155
                     /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             2552..3361
                     /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(2419..2439)
                     /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(2396..2415)
                     /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    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"
     intron          2248..2336
                     /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    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    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    2339..2345
                     /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(2233..2239)
                     /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    complement(2229..2231)
                     /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"
     misc_feature    2347..2349
                     /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        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"
     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"
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 AACTTAATAA GCTGAACCCT CCTGATGAGA GTGGCCCCGG CTGCATGAGC TGCAAGTGTG
     1861 TGCTCTCCTG AGGCGCGCCT CTAGAACTAT AGTGAGTCGT ATTACGTAGA TCCAGACATG
     1921 ATAAGATACA TTGATGAGTT TGGACAAACC ACAACTAGAA TGCAGTGAAA AAAATGCTTT
     1981 ATTTGTGAAA TTTGTGATGC TATTGCTTTA TTTGTAACCA TTATAAGCTG CAATAAACAA
     2041 GTTAACAACA ACAATTGCAT TCATTTTATG TTTCAGGTTC AGGGGGAGGT GTGGGAGGTT
     2101 TTTTCCAACT TTATTATACA AAGTTGGCAT TATAAAAAAG CATTGCTTAT CAATTTGTTG
     2161 CAACGAACAG GTCACTATCA GTCAAAATAA AATCATTATT TGGAGCTCCA TGGcgttgtc
     2221 tagcaaggaa gtgaagagct atccggatat atatgtatgt tatcataact gtactggcta
     2281 tgtcaggtgg taatgactgt aacgttacgt tactcgttgt aggcacgaca ttgaatatgc
     2341 tcttctCCGC TGTGGCCCGA ACGCCTCCCg ctagcgatgg gatctCGGCC GCGATATCCC
     2401 CTATAGTGAG TCGTATTACA TGGTCATAGC TGTTTCCTGG CAGCTCTGGC CCGTGTCTCA
     2461 AAATCTCTGA TGTTACATTG CACAAGATAA AAATATATCA TCATGAACAA TAAAACTGTC
     2521 TGCTTACATA AACAGTAATA CAAGGGGTGT TATGAGCCAT ATTCAACGGG AAACGTCGAG
     2581 GCCGCGATTA AATTCCAACA TGGATGCTGA TTTATATGGG TATAAATGGG CTCGCGATAA
     2641 TGTCGGGCAA TCAGGTGCGA CAATCTATCG CTTGTATGGG AAGCCCGATG CGCCAGAGTT
     2701 GTTTCTGAAA CATGGCAAAG GTAGCGTTGC CAATGATGTT ACAGATGAGA TGGTCAGACT
     2761 AAACTGGCTG ACGGAATTTA TGCCTCTTCC GACCATCAAG CATTTTATCC GTACTCCTGA
     2821 TGATGCATGG TTACTCACCA CTGCGATCCC CGGAAAAACA GCATTCCAGG TATTAGAAGA
     2881 ATATCCTGAT TCAGGTGAAA ATATTGTTGA TGCGCTGGCA GTGTTCCTGC GCCGGTTGCA
     2941 TTCGATTCCT GTTTGTAATT GTCCTTTTAA CAGCGATCGC GTATTTCGTC TCGCTCAGGC
     3001 GCAATCACGA ATGAATAACG GTTTGGTTGA TGCGAGTGAT TTTGATGACG AGCGTAATGG
     3061 CTGGCCTGTT GAACAAGTCT GGAAAGAAAT GCATAAACTT TTGCCATTCT CACCGGATTC
     3121 AGTCGTCACT CATGGTGATT TCTCACTTGA TAACCTTATT TTTGACGAGG GGAAATTAAT
     3181 AGGTTGTATT GATGTTGGAC GAGTCGGAAT CGCAGACCGA TACCAGGATC TTGCCATCCT
     3241 ATGGAACTGC CTCGGTGAGT TTTCTCCTTC ATTACAGAAA CGGCTTTTTC AAAAATATGG
     3301 TATTGATAAT CCTGATATGA ATAAATTGCA GTTTCATTTG ATGCTCGATG AGTTTTTCTA
     3361 ATCAGAATTG GTTAATTGGT TGTAACACTG GCAGAGCATT ACGCTGACTT GACGGGACGG
     3421 CGCAAGCTCA TGACCAAAAT CCCTTAACGT GAGTTACGCG TCGTTCCACT GAGCGTCAGA
     3481 CCCCGTAGAA AAGATCAAAG GATCTTCTTG AGATCCTTTT TTTCTGCGCG TAATCTGCTG
     3541 CTTGCAAACA AAAAAACCAC CGCTACCAGC GGTGGTTTGT TTGCCGGATC AAGAGCTACC
     3601 AACTCTTTTT CCGAAGGTAA CTGGCTTCAG CAGAGCGCAG ATACCAAATA CTGTCCTTCT
     3661 AGTGTAGCCG TAGTTAGGCC ACCACTTCAA GAACTCTGTA GCACCGCCTA CATACCTCGC
     3721 TCTGCTAATC CTGTTACCAG TGGCTGCTGC CAGTGGCGAT AAGTCGTGTC TTACCGGGTT
     3781 GGACTCAAGA CGATAGTTAC CGGATAAGGC GCAGCGGTCG GGCTGAACGG GGGGTTCGTG
     3841 CACACAGCCC AGCTTGGAGC GAACGACCTA CACCGAACTG AGATACCTAC AGCGTGAGCA
     3901 TTGAGAAAGC GCCACGCTTC CCGAAGGGAG AAAGGCGGAC AGGTATCCGG TAAGCGGCAG
     3961 GGTCGGAACA GGAGAGCGCA CGAGGGAGCT TCCAGGGGGA AACGCCTGGT ATCTTTATAG
     4021 TCCTGTCGGG TTTCGCCACC TCTGACTTGA GCGTCGATTT TTGTGATGCT CGTCAGGGGG
     4081 GCGGAGCCTA TGGAAAAACG CCAGCAACGC GGCCTTTTTA CGGTTCCTGG CCTTTTGCTG
     4141 GCCTTTTGCT CACATGTT    
//
