LOCUS       pGTag_NLS_TagRFP_B_actin        4801 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      4125..4798
                     /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             3195..4004
                     /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(3062..3082)
                     /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(3039..3058)
                     /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(2990..3012)
                     /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(2876..2882)
                     /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(2872..2874)
                     /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          2891..2979
                     /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    2990..2992
                     /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    2982..2988
                     /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_structure  1098..1124
                     /locus_tag="NLS"
                     /ApEinfo_fwdcolor="#ff0000"
                     /ApEinfo_revcolor="#ff0000"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
     CDS             1125..1835
                     /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    1847..2445
                     /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    2426..2850
                     /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 GCTCCAAAGA AGAAGCGTAA GGTAATGGTG TCTAAGGGCG
     1141 AgGAaCTGAT TAAGGAGAAC ATGCACATGA AGCTGTACAT GGAGGGCACC GTGAACAACC
     1201 ACCACTTCAA GTGCACATCC GAGGGCGAAG GCAAGCCCTA CGAGGGCACC CAGACCATGA
     1261 GAATCAAGGT GGTCGAGGGC GGCCCTCTCC CCTTCGCCTT CGACATCCTG GCTACCAGCT
     1321 TCATGTACGG CAGCAGAACC TTCATCAACC ACACCCAGGG CATCCCCGAC TTCTTTAAGC
     1381 AGTCCTTCCC TGAGGGCTTC ACATGGGAGA GAGTCACCAC ATACGAAGAC GGGGGCGTGC
     1441 TGACCGCTAC CCAGGACACC AGCCTCCAGG ACGGCTGCCT CATCTACAAC GTCAAGATCA
     1501 GAGGGGTGAA CTTCCCATCC AACGGCCCTG TGATGCAGAA GAAAACACTC GGCTGGGAGG
     1561 CCAACACCGA GATGCTGTAC CCCGCTGACG GCGGCCTGGA AGGCAGAAGC GACATGGCCC
     1621 TGAAGCTCGT GGGCGGGGGC CACCTGATCT GCAACTTCAA GACCACATAC AGATCCAAGA
     1681 AACCCGCTAA GAACCTCAAG ATGCCCGGCG TCTACTATGT GGACCACAGA CTGGAAAGAA
     1741 TCAAGGAGGC CGACAAAGAG ACCTACGTCG AGCAGCACGA GGTGGCTGTG GCCAGATACT
     1801 GCGACCTCCC TAGCAAACTG GGGCACAAAC TTAATTGAGG CGCGCCACGG ACTGTTACCA
     1861 CTTCACGCCG ACTCAAACTG CGCAGAGAAA CTTCAAACGA CAACATTGGC ATGGCTTTTG
     1921 TTATTTTTGG CGCTTGACTC AGGATCTAAA AACTGGAACG GTGAAGGTGA CGGCAATGTT
     1981 TTTTGGCAAA TAAGCATCCC CGAAGTTCTA CAATGCATCT GAGGACTCCA TGTTTTGTTT
     2041 TCTTTAGTCA TTCCAAATGT TTGTTAAATG CATTGTTCCG AAACTTATTT GCCTCTATGA
     2101 AGGCTGCCCA GTAATTGGGA GCATACTTAA CATTGTAGTA TTGTATGTAA ATTATGTAAC
     2161 AAAACAATGT CTGGGTTTTT GTACTTTTCA GCCTTAAAAT CTTGGGTTTA TTTTTTTTTC
     2221 TTTGTTCCAA AAAAACGAAG CTTTACCATT CAAGATGTAA AGGTTTCCAT TCCCCCTGGG
     2281 CATATTGTAA AAGCTGTGTG GAACGTGGCG GTGCCAGACA TTTGGTGGGG CCAACCTGTA
     2341 CACTGACTAA TTCAATTCCA ATAAAAGTGC ACATGTAAGA CATCCTAcCC CTGTCTCATT
     2401 TCCTGTCTTG GCTCTGAGTG GCTTCAATAA AAATGGCTTT TGTGAAGTTG CACCATGtTA
     2461 ATTACATTTC AGCACTTTCA CACCAGGTTT TTGACTAATT TGAATTTATC GAAGTTTGAC
     2521 CTTTTTTTTT TTTTTTCTTC AAGCTAATTT GAGGTTATCA AACCATTTCA TTGGAGGAAA
     2581 CTTAATGTTT AAAAGCATAA ATTCGAAACA TGATTCATTT ATTTAACTGC TTAATGGTGA
     2641 AACAAACTTC AAGTAATACA CCAAGTGTTC CCTCATATGA AGGTAGCTAA CTGCAAACGG
     2701 CCTTAACTTT CCATCTTAAG TTGAGCTACA TAAAGGCTTT AACAGGAAGG AActGCCAGT
     2761 TTAAAAGTCA AACCACCATG ACTGATATAA ATTCTTGTCC ATATAACCTT GCGTTTCACA
     2821 CAATATAGCT ATCTACTGCT AAATAAATTA CCATGGcgtt gtctagcaag gaagtgaaga
     2881 gctatccgga tatatatgta tgttatcata actgtactgg ctatgtcagg tggtaatgac
     2941 tgtaacgtta cgttactcgt tgtaggcacg acattgaata tgctcttctC CGCTGTGGCC
     3001 CGAACGCCTC CCgctagcga tgggatctCG GCCGCGATAT CCCCTATAGT GAGTCGTATT
     3061 ACATGGTCAT AGCTGTTTCC TGGCAGCTCT GGCCCGTGTC TCAAAATCTC TGATGTTACA
     3121 TTGCACAAGA TAAAAATATA TCATCATGAA CAATAAAACT GTCTGCTTAC ATAAACAGTA
     3181 ATACAAGGGG TGTTATGAGC CATATTCAAC GGGAAACGTC GAGGCCGCGA TTAAATTCCA
     3241 ACATGGATGC TGATTTATAT GGGTATAAAT GGGCTCGCGA TAATGTCGGG CAATCAGGTG
     3301 CGACAATCTA TCGCTTGTAT GGGAAGCCCG ATGCGCCAGA GTTGTTTCTG AAACATGGCA
     3361 AAGGTAGCGT TGCCAATGAT GTTACAGATG AGATGGTCAG ACTAAACTGG CTGACGGAAT
     3421 TTATGCCTCT TCCGACCATC AAGCATTTTA TCCGTACTCC TGATGATGCA TGGTTACTCA
     3481 CCACTGCGAT CCCCGGAAAA ACAGCATTCC AGGTATTAGA AGAATATCCT GATTCAGGTG
     3541 AAAATATTGT TGATGCGCTG GCAGTGTTCC TGCGCCGGTT GCATTCGATT CCTGTTTGTA
     3601 ATTGTCCTTT TAACAGCGAT CGCGTATTTC GTCTCGCTCA GGCGCAATCA CGAATGAATA
     3661 ACGGTTTGGT TGATGCGAGT GATTTTGATG ACGAGCGTAA TGGCTGGCCT GTTGAACAAG
     3721 TCTGGAAAGA AATGCATAAA CTTTTGCCAT TCTCACCGGA TTCAGTCGTC ACTCATGGTG
     3781 ATTTCTCACT TGATAACCTT ATTTTTGACG AGGGGAAATT AATAGGTTGT ATTGATGTTG
     3841 GACGAGTCGG AATCGCAGAC CGATACCAGG ATCTTGCCAT CCTATGGAAC TGCCTCGGTG
     3901 AGTTTTCTCC TTCATTACAG AAACGGCTTT TTCAAAAATA TGGTATTGAT AATCCTGATA
     3961 TGAATAAATT GCAGTTTCAT TTGATGCTCG ATGAGTTTTT CTAATCAGAA TTGGTTAATT
     4021 GGTTGTAACA CTGGCAGAGC ATTACGCTGA CTTGACGGGA CGGCGCAAGC TCATGACCAA
     4081 AATCCCTTAA CGTGAGTTAC GCGTCGTTCC ACTGAGCGTC AGACCCCGTA GAAAAGATCA
     4141 AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA ACAAAAAAAC
     4201 CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT TTTCCGAAGG
     4261 TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTCCT TCTAGTGTAG CCGTAGTTAG
     4321 GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA ATCCTGTTAC
     4381 CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA AGACGATAGT
     4441 TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG CCCAGCTTGG
     4501 AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA GCATTGAGAA AGCGCCACGC
     4561 TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC
     4621 GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC GGGTTTCGCC
     4681 ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC CTATGGAAAA
     4741 ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT GCTCACATGT
     4801 T     
//
