LOCUS       pGTag_eGFP_caax_B_actin        4840 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_feature    complement(3028..3050)
                     /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_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      4163..4836
                     /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             3233..4042
                     /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(3100..3120)
                     /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(2914..2920)
                     /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    3028..3030
                     /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(3077..3096)
                     /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          2929..3017
                     /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    3020..3026
                     /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(2910..2912)
                     /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    1885..2483
                     /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    2464..2888
                     /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"
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 CGCCACGGAC TGTTACCACT TCACGCCGAC TCAAACTGCG
     1921 CAGAGAAACT TCAAACGACA ACATTGGCAT GGCTTTTGTT ATTTTTGGCG CTTGACTCAG
     1981 GATCTAAAAA CTGGAACGGT GAAGGTGACG GCAATGTTTT TTGGCAAATA AGCATCCCCG
     2041 AAGTTCTACA ATGCATCTGA GGACTCCATG TTTTGTTTTC TTTAGTCATT CCAAATGTTT
     2101 GTTAAATGCA TTGTTCCGAA ACTTATTTGC CTCTATGAAG GCTGCCCAGT AATTGGGAGC
     2161 ATACTTAACA TTGTAGTATT GTATGTAAAT TATGTAACAA AACAATGTCT GGGTTTTTGT
     2221 ACTTTTCAGC CTTAAAATCT TGGGTTTATT TTTTTTTCTT TGTTCCAAAA AAACGAAGCT
     2281 TTACCATTCA AGATGTAAAG GTTTCCATTC CCCCTGGGCA TATTGTAAAA GCTGTGTGGA
     2341 ACGTGGCGGT GCCAGACATT TGGTGGGGCC AACCTGTACA CTGACTAATT CAATTCCAAT
     2401 AAAAGTGCAC ATGTAAGACA TCCTAcCCCT GTCTCATTTC CTGTCTTGGC TCTGAGTGGC
     2461 TTCAATAAAA ATGGCTTTTG TGAAGTTGCA CCATGtTAAT TACATTTCAG CACTTTCACA
     2521 CCAGGTTTTT GACTAATTTG AATTTATCGA AGTTTGACCT TTTTTTTTTT TTTTCTTCAA
     2581 GCTAATTTGA GGTTATCAAA CCATTTCATT GGAGGAAACT TAATGTTTAA AAGCATAAAT
     2641 TCGAAACATG ATTCATTTAT TTAACTGCTT AATGGTGAAA CAAACTTCAA GTAATACACC
     2701 AAGTGTTCCC TCATATGAAG GTAGCTAACT GCAAACGGCC TTAACTTTCC ATCTTAAGTT
     2761 GAGCTACATA AAGGCTTTAA CAGGAAGGAA ctGCCAGTTT AAAAGTCAAA CCACCATGAC
     2821 TGATATAAAT TCTTGTCCAT ATAACCTTGC GTTTCACACA ATATAGCTAT CTACTGCTAA
     2881 ATAAATTACC ATGGcgttgt ctagcaagga agtgaagagc tatccggata tatatgtatg
     2941 ttatcataac tgtactggct atgtcaggtg gtaatgactg taacgttacg ttactcgttg
     3001 taggcacgac attgaatatg ctcttctCCG CTGTGGCCCG AACGCCTCCC gctagcgatg
     3061 ggatctCGGC CGCGATATCC CCTATAGTGA GTCGTATTAC ATGGTCATAG CTGTTTCCTG
     3121 GCAGCTCTGG CCCGTGTCTC AAAATCTCTG ATGTTACATT GCACAAGATA AAAATATATC
     3181 ATCATGAACA ATAAAACTGT CTGCTTACAT AAACAGTAAT ACAAGGGGTG TTATGAGCCA
     3241 TATTCAACGG GAAACGTCGA GGCCGCGATT AAATTCCAAC ATGGATGCTG ATTTATATGG
     3301 GTATAAATGG GCTCGCGATA ATGTCGGGCA ATCAGGTGCG ACAATCTATC GCTTGTATGG
     3361 GAAGCCCGAT GCGCCAGAGT TGTTTCTGAA ACATGGCAAA GGTAGCGTTG CCAATGATGT
     3421 TACAGATGAG ATGGTCAGAC TAAACTGGCT GACGGAATTT ATGCCTCTTC CGACCATCAA
     3481 GCATTTTATC CGTACTCCTG ATGATGCATG GTTACTCACC ACTGCGATCC CCGGAAAAAC
     3541 AGCATTCCAG GTATTAGAAG AATATCCTGA TTCAGGTGAA AATATTGTTG ATGCGCTGGC
     3601 AGTGTTCCTG CGCCGGTTGC ATTCGATTCC TGTTTGTAAT TGTCCTTTTA ACAGCGATCG
     3661 CGTATTTCGT CTCGCTCAGG CGCAATCACG AATGAATAAC GGTTTGGTTG ATGCGAGTGA
     3721 TTTTGATGAC GAGCGTAATG GCTGGCCTGT TGAACAAGTC TGGAAAGAAA TGCATAAACT
     3781 TTTGCCATTC TCACCGGATT CAGTCGTCAC TCATGGTGAT TTCTCACTTG ATAACCTTAT
     3841 TTTTGACGAG GGGAAATTAA TAGGTTGTAT TGATGTTGGA CGAGTCGGAA TCGCAGACCG
     3901 ATACCAGGAT CTTGCCATCC TATGGAACTG CCTCGGTGAG TTTTCTCCTT CATTACAGAA
     3961 ACGGCTTTTT CAAAAATATG GTATTGATAA TCCTGATATG AATAAATTGC AGTTTCATTT
     4021 GATGCTCGAT GAGTTTTTCT AATCAGAATT GGTTAATTGG TTGTAACACT GGCAGAGCAT
     4081 TACGCTGACT TGACGGGACG GCGCAAGCTC ATGACCAAAA TCCCTTAACG TGAGTTACGC
     4141 GTCGTTCCAC TGAGCGTCAG ACCCCGTAGA AAAGATCAAA GGATCTTCTT GAGATCCTTT
     4201 TTTTCTGCGC GTAATCTGCT GCTTGCAAAC AAAAAAACCA CCGCTACCAG CGGTGGTTTG
     4261 TTTGCCGGAT CAAGAGCTAC CAACTCTTTT TCCGAAGGTA ACTGGCTTCA GCAGAGCGCA
     4321 GATACCAAAT ACTGTCCTTC TAGTGTAGCC GTAGTTAGGC CACCACTTCA AGAACTCTGT
     4381 AGCACCGCCT ACATACCTCG CTCTGCTAAT CCTGTTACCA GTGGCTGCTG CCAGTGGCGA
     4441 TAAGTCGTGT CTTACCGGGT TGGACTCAAG ACGATAGTTA CCGGATAAGG CGCAGCGGTC
     4501 GGGCTGAACG GGGGGTTCGT GCACACAGCC CAGCTTGGAG CGAACGACCT ACACCGAACT
     4561 GAGATACCTA CAGCGTGAGC ATTGAGAAAG CGCCACGCTT CCCGAAGGGA GAAAGGCGGA
     4621 CAGGTATCCG GTAAGCGGCA GGGTCGGAAC AGGAGAGCGC ACGAGGGAGC TTCCAGGGGG
     4681 AAACGCCTGG TATCTTTATA GTCCTGTCGG GTTTCGCCAC CTCTGACTTG AGCGTCGATT
     4741 TTTGTGATGC TCGTCAGGGG GGCGGAGCCT ATGGAAAAAC GCCAGCAACG CGGCCTTTTT
     4801 ACGGTTCCTG GCCTTTTGCT GGCCTTTTGC TCACATGTTC  
//
