LOCUS       Exported                4390 bp ds-DNA     circular SYN 29-JUN-2020
DEFINITION  synthetic circular DNA, gene-targeting cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4390)
  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
REFERENCE   2  (bases 1 to 4390)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 29, 2020 from SnapGene Viewer 5.1.4.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4390
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      268..295
                     /locus_tag="T2"
                     /label=T2 terminator
     terminator      427..470
                     /locus_tag="T1"
                     /label=T1 terminator
     primer_bind     537..552
                     /locus_tag="M13F"
                     /label=M13F
     misc_feature    591..613
                     /locus_tag="5' universal guide (w/PAM)"
                     /label=5' universal guide (w/PAM)
     misc_feature    complement(610..613)
                     /locus_tag="BfuAI site 1 overhang"
                     /label=BfuAI site 1 overhang
     misc_feature    complement(618..623)
                     /locus_tag="BfuAI site 1"
                     /label=BfuAI site 1
     promoter        624..745
                     /locus_tag="lacZ\promoter"
                     /label=lacZ promoter
                     /note="/vntifkey=30"
     misc_feature    746..1021
                     /locus_tag="lacZ'"
                     /label=lacZ'
                     /note="/vntifkey=21"
     misc_feature    1022..1027
                     /locus_tag="BfuAI site 2"
                     /label=BfuAI site 2
     misc_feature    1032..1037
                     /locus_tag="PTV1-2A peptide(1)"
                     /label=PTV1-2A peptide
     misc_feature    1032..1037
                     /locus_tag="BamHI"
                     /label=BamHI
     misc_feature    1032..1035
                     /locus_tag="BfuAI site 2 overhang"
                     /label=BfuAI site 2 overhang
     misc_feature    1038..1094
                     /locus_tag="PTV1-2A peptide"
                     /label=PTV1-2A peptide
     misc_feature    1095..1100
                     /locus_tag="MCS"
                     /label=MCS
     misc_feature    1095..1100
                     /locus_tag="XmaI"
                     /label=XmaI
     misc_feature    1101..1124
                     /locus_tag="NLS"
                     /label=NLS
     misc_feature    1125..1679
                     /locus_tag="QFDBD"
                     /label=QFDBD
     misc_feature    1686..1691
                     /locus_tag="SpeI"
                     /label=SpeI
     misc_feature    1701..2102
                     /locus_tag="Gal4AD"
                     /label=Gal4AD
     misc_feature    2104..2111
                     /locus_tag="AscI"
                     /label=AscI
     polyA_signal    2141..2336
                     /locus_tag="SV40 late polyA (from pCS2+)"
                     /label=SV40 late polyA (from pCS2+)
     misc_recomb     2338..2433
                     /locus_tag="attL3"
                     /label=attL3
     misc_feature    2338..2352
                     /locus_tag="att3_shared"
                     /label=att3_shared
     misc_feature    complement(2461..2463)
                     /locus_tag="BspQI site 1 overhang"
                     /label=BspQI site 1 overhang
     misc_feature    complement(2465..2471)
                     /locus_tag="BspQI site 1"
                     /label=BspQI site 1
     intron          2480..2568
                     /locus_tag="Carp beta-actin intron 1 filler"
                     /label=Carp beta-actin intron 1 filler
                     /note="Carp beta-actin intron 1 filler"
     misc_feature    2571..2577
                     /locus_tag="BspQI site 2"
                     /label=BspQI site 2
     misc_feature    complement(2579..2601)
                     /locus_tag="3' Universal Guide (w/PAM)"
                     /label=3' Universal Guide (w/PAM)
     misc_feature    2579..2581
                     /locus_tag="BspQI site 2 overhang"
                     /label=BspQI site 2 overhang
     promoter        complement(2628..2647)
                     /locus_tag="T7"
                     /label=T7 promoter
     primer_bind     complement(2651..2671)
                     /locus_tag="M13R"
                     /label=M13R
     CDS             2784..3593
                     /locus_tag="kanR"
                     /label=kanR
     rep_origin      3714..4387
                     /locus_tag="pUC ori"
                     /label=pUC ori
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 cggtcccggg atggcaccca agaagaagag gaagatgccg cctaaacgca
     1141 agacactcaa tgccgctgcc gaagccaatg cccacgctga tggccatgct gatggcaatg
     1201 ctgatggtca tgtcgctaac actgcagcaa gcagcaacaa cgcccgtttt gcggacttga
     1261 ccaacattga cacacccggc ctcggcccta ccacgacgac gttacttgtc gagcccgctc
     1321 gttcgaaacg ccagagagtc tcgagggcct gtgatcagtg tcgagctgca cgtgaaaagt
     1381 gtgatggaat ccagccggct tgcttcccct gtgtgtcgca gggccggtcg tgtacctacc
     1441 aggccagtcc caagaagcga ggagtccaga cgggctacat ccgcactctc gaactggctc
     1501 tggcttggat gttcgagaac gttgcccgca gcgaggacgc cctccacaat cttttggtcc
     1561 gtgatgctgg ccagggcagc gctctcctgg tcggcaaaga ctcgcctgct gcagaacgcc
     1621 tgcatgcaag atgggcgacg agtcgagtca acaaaagcat cacccgtctt ctctcaggtg
     1681 ctagcactag tagcgccggt caacagcaac agctgcaatc attagtgcca ctgaccccgt
     1741 ctgctttgtt tggtggcgcc aattttaatc aaagtgggaa tattgctgat agctcattgt
     1801 ccttcacttt cactaacagt agcaacggtc cgaacctcat aacaactcaa acaaattctc
     1861 aagcgctttc acaaccaatt gcctcctcta acgttcatga taacttcatg aataatgaaa
     1921 tcacggctag taaaattgat gatggtaata attcaaaacc actgtcacct ggttggacgg
     1981 accaaactgc gtataacgcg tttggaatca ctacagggat gtttaatacc actacaatgg
     2041 atgatgtata taactatcta ttcgatgatg aagatacccc accaaaccca aaaaaagagt
     2101 aaaggcgcgc ctctagaact atagtgagtc gtattacgta gatccagaca tgataagata
     2161 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2221 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2281 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttccaa
     2341 ctttattata caaagttggc attataaaaa agcattgctt atcaatttgt tgcaacgaac
     2401 aggtcactat cagtcaaaat aaaatcatta tttggagctc catggcgttg tctagcaagg
     2461 aagtgaagag ctatccggat atatatgtat gttatcataa ctgtactggc tatgtcaggt
     2521 ggtaatgact gtaacgttac gttactcgtt gtaggcacga cattgaatat gctcttctcc
     2581 gctgtggccc gaacgcctcc cgctagcgat gggatctcgg ccgcgatatc ccctatagtg
     2641 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     2701 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     2761 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     2821 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     2881 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     2941 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     3001 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     3061 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     3121 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     3181 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     3241 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     3301 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     3361 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     3421 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     3481 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     3541 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     3601 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     3661 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     3721 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3781 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3841 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     3901 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3961 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4021 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4081 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa
     4141 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4201 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4261 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4321 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4381 ctcacatgtt
//
