LOCUS       pGTag_TagRFP_B_actin        4774 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      4098..4771
                     /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             3168..3977
                     /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(3035..3055)
                     /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(3012..3031)
                     /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(2963..2985)
                     /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(2849..2855)
                     /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(2845..2847)
                     /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          2864..2952
                     /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    2963..2965
                     /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    2955..2961
                     /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"
     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    1820..2418
                     /locus_tag="Danio b-Actin 3' UTR"
                     /ApEinfo_fwdcolor="#0000ff"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     misc_feature    2399..2823
                     /locus_tag="Danio b-Actin poly(A) putative"
                     /ApEinfo_fwdcolor="#004080"
                     /ApEinfo_revcolor="#004080"
                     /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 AGGCGCGCCA CGGACTGTTA CCACTTCACG CCGACTCAAA CTGCGCAGAG
     1861 AAACTTCAAA CGACAACATT GGCATGGCTT TTGTTATTTT TGGCGCTTGA CTCAGGATCT
     1921 AAAAACTGGA ACGGTGAAGG TGACGGCAAT GTTTTTTGGC AAATAAGCAT CCCCGAAGTT
     1981 CTACAATGCA TCTGAGGACT CCATGTTTTG TTTTCTTTAG TCATTCCAAA TGTTTGTTAA
     2041 ATGCATTGTT CCGAAACTTA TTTGCCTCTA TGAAGGCTGC CCAGTAATTG GGAGCATACT
     2101 TAACATTGTA GTATTGTATG TAAATTATGT AACAAAACAA TGTCTGGGTT TTTGTACTTT
     2161 TCAGCCTTAA AATCTTGGGT TTATTTTTTT TTCTTTGTTC CAAAAAAACG AAGCTTTACC
     2221 ATTCAAGATG TAAAGGTTTC CATTCCCCCT GGGCATATTG TAAAAGCTGT GTGGAACGTG
     2281 GCGGTGCCAG ACATTTGGTG GGGCCAACCT GTACACTGAC TAATTCAATT CCAATAAAAG
     2341 TGCACATGTA AGACATCCTA cCCCTGTCTC ATTTCCTGTC TTGGCTCTGA GTGGCTTCAA
     2401 TAAAAATGGC TTTTGTGAAG TTGCACCATG tTAATTACAT TTCAGCACTT TCACACCAGG
     2461 TTTTTGACTA ATTTGAATTT ATCGAAGTTT GACCTTTTTT TTTTTTTTTC TTCAAGCTAA
     2521 TTTGAGGTTA TCAAACCATT TCATTGGAGG AAACTTAATG TTTAAAAGCA TAAATTCGAA
     2581 ACATGATTCA TTTATTTAAC TGCTTAATGG TGAAACAAAC TTCAAGTAAT ACACCAAGTG
     2641 TTCCCTCATA TGAAGGTAGC TAACTGCAAA CGGCCTTAAC TTTCCATCTT AAGTTGAGCT
     2701 ACATAAAGGC TTTAACAGGA AGGAActGCC AGTTTAAAAG TCAAACCACC ATGACTGATA
     2761 TAAATTCTTG TCCATATAAC CTTGCGTTTC ACACAATATA GCTATCTACT GCTAAATAAA
     2821 TTACCATGGc gttgtctagc aaggaagtga agagctatcc ggatatatat gtatgttatc
     2881 ataactgtac tggctatgtc aggtggtaat gactgtaacg ttacgttact cgttgtaggc
     2941 acgacattga atatgctctt ctCCGCTGTG GCCCGAACGC CTCCCgctag cgatgggatc
     3001 tCGGCCGCGA TATCCCCTAT AGTGAGTCGT ATTACATGGT CATAGCTGTT TCCTGGCAGC
     3061 TCTGGCCCGT GTCTCAAAAT CTCTGATGTT ACATTGCACA AGATAAAAAT ATATCATCAT
     3121 GAACAATAAA ACTGTCTGCT TACATAAACA GTAATACAAG GGGTGTTATG AGCCATATTC
     3181 AACGGGAAAC GTCGAGGCCG CGATTAAATT CCAACATGGA TGCTGATTTA TATGGGTATA
     3241 AATGGGCTCG CGATAATGTC GGGCAATCAG GTGCGACAAT CTATCGCTTG TATGGGAAGC
     3301 CCGATGCGCC AGAGTTGTTT CTGAAACATG GCAAAGGTAG CGTTGCCAAT GATGTTACAG
     3361 ATGAGATGGT CAGACTAAAC TGGCTGACGG AATTTATGCC TCTTCCGACC ATCAAGCATT
     3421 TTATCCGTAC TCCTGATGAT GCATGGTTAC TCACCACTGC GATCCCCGGA AAAACAGCAT
     3481 TCCAGGTATT AGAAGAATAT CCTGATTCAG GTGAAAATAT TGTTGATGCG CTGGCAGTGT
     3541 TCCTGCGCCG GTTGCATTCG ATTCCTGTTT GTAATTGTCC TTTTAACAGC GATCGCGTAT
     3601 TTCGTCTCGC TCAGGCGCAA TCACGAATGA ATAACGGTTT GGTTGATGCG AGTGATTTTG
     3661 ATGACGAGCG TAATGGCTGG CCTGTTGAAC AAGTCTGGAA AGAAATGCAT AAACTTTTGC
     3721 CATTCTCACC GGATTCAGTC GTCACTCATG GTGATTTCTC ACTTGATAAC CTTATTTTTG
     3781 ACGAGGGGAA ATTAATAGGT TGTATTGATG TTGGACGAGT CGGAATCGCA GACCGATACC
     3841 AGGATCTTGC CATCCTATGG AACTGCCTCG GTGAGTTTTC TCCTTCATTA CAGAAACGGC
     3901 TTTTTCAAAA ATATGGTATT GATAATCCTG ATATGAATAA ATTGCAGTTT CATTTGATGC
     3961 TCGATGAGTT TTTCTAATCA GAATTGGTTA ATTGGTTGTA ACACTGGCAG AGCATTACGC
     4021 TGACTTGACG GGACGGCGCA AGCTCATGAC CAAAATCCCT TAACGTGAGT TACGCGTCGT
     4081 TCCACTGAGC GTCAGACCCC GTAGAAAAGA TCAAAGGATC TTCTTGAGAT CCTTTTTTTC
     4141 TGCGCGTAAT CTGCTGCTTG CAAACAAAAA AACCACCGCT ACCAGCGGTG GTTTGTTTGC
     4201 CGGATCAAGA GCTACCAACT CTTTTTCCGA AGGTAACTGG CTTCAGCAGA GCGCAGATAC
     4261 CAAATACTGT CCTTCTAGTG TAGCCGTAGT TAGGCCACCA CTTCAAGAAC TCTGTAGCAC
     4321 CGCCTACATA CCTCGCTCTG CTAATCCTGT TACCAGTGGC TGCTGCCAGT GGCGATAAGT
     4381 CGTGTCTTAC CGGGTTGGAC TCAAGACGAT AGTTACCGGA TAAGGCGCAG CGGTCGGGCT
     4441 GAACGGGGGG TTCGTGCACA CAGCCCAGCT TGGAGCGAAC GACCTACACC GAACTGAGAT
     4501 ACCTACAGCG TGAGCATTGA GAAAGCGCCA CGCTTCCCGA AGGGAGAAAG GCGGACAGGT
     4561 ATCCGGTAAG CGGCAGGGTC GGAACAGGAG AGCGCACGAG GGAGCTTCCA GGGGGAAACG
     4621 CCTGGTATCT TTATAGTCCT GTCGGGTTTC GCCACCTCTG ACTTGAGCGT CGATTTTTGT
     4681 GATGCTCGTC AGGGGGGCGG AGCCTATGGA AAAACGCCAG CAACGCGGCC TTTTTACGGT
     4741 TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTT  
//
