LOCUS       pU6-(BbsI)_CBh-Cas9-T2A-BFP      9292 bp    DNA     circular UNA 09-MAY-2014
DEFINITION  .
ACCESSION   
VERSION     
SOURCE      . (unknown)
  ORGANISM  .
            .
COMMENT     
FEATURES             Location/Qualifiers
     promoter        1..249
                     /label=hU6
     Site            252..255
                     /label=BbsI
     Site            274..277
                     /label=BbsI
     misc_feature    274..355
                     /label="Guide RNA"
                     /label="sgRNA scaffold"
     promoter        440..1238
                     /label=CBh
                     /label="CBh promoter"
     CDS             1251..1319
                     /label=3xFLAG
     CDS             1320..1370
                     /label=NLS
     CDS             1371..5471
                     /label=hSpCas9
     CDS             5472..5519
                     /label=NLS(1)
     CDS             5529..5583
                     /label=T2A
     CDS             5589..6308
                     /label=BFP
     polyA_signal    6309..6540
                     /label=pGHpA
                     /label=bpA
     CDS             7612..8472
                     /label=ampR
ORIGIN
        1 CCCCACGAGG GCCTATTTCC CATGATTCCT TCATATTTGC ATATACGATA CAAGGCTGTT
       61 AGAGAGATAA TTGGAATTAA TTTGACTGTA AACACAAAGA TATTAGTACA AAATACGTGA
      121 CGTAGAAAGT AATAATTTCT TGGGTAGTTT GCAGTTTTAA AATTATGTTT TAAAATGGAC
      181 TATCATATGC TTACCGTAAC TTGAAAGTAT TTCGATTTCT TGGCTTTATA TATCTTGTGG
      241 AAAGGACGAA ACACCGGGTC TTCGAGAAGA CCTGTTTTAG AGCTAGAAAT AGCAAGTTAA
      301 AATAAGGCTA GTCCGTTATC AACTTGAAAA AGTGGCACCG AGTCGGTGCT TTTTTGTTTT
      361 AGAGCTAGAA ATAGCAAGTT AAAATAAGGC TAGTCCGTTT TTAGCGCGTG CGCCAATTCT
      421 GCAGACAAAT GGCTCTAGAG GTACCCGTTA CATAACTTAC GGTAAATGGC CCGCCTGGCT
      481 GACCGCCCAA CGACCCCCGC CCATTGACGT CAATAGTAAC GCCAATAGGG ACTTTCCATT
      541 GACGTCAATG GGTGGAGTAT TTACGGTAAA CTGCCCACTT GGCAGTACAT CAAGTGTATC
      601 ATATGCCAAG TACGCCCCCT ATTGACGTCA ATGACGGTAA ATGGCCCGCC TGGCATTGTG
      661 CCCAGTACAT GACCTTATGG GACTTTCCTA CTTGGCAGTA CATCTACGTA TTAGTCATCG
      721 CTATTACCAT GGTCGAGGTG AGCCCCACGT TCTGCTTCAC TCTCCCCATC TCCCCCCCCT
      781 CCCCACCCCC AATTTTGTAT TTATTTATTT TTTAATTATT TTGTGCAGCG ATGGGGGCGG
      841 GGGGGGGGGG GGGGCGCGCG CCAGGCGGGG CGGGGCGGGG CGAGGGGCGG GGCGGGGCGA
      901 GGCGGAGAGG TGCGGCGGCA GCCAATCAGA GCGGCGCGCT CCGAAAGTTT CCTTTTATGG
      961 CGAGGCGGCG GCGGCGGCGG CCCTATAAAA AGCGAAGCGC GCGGCGGGCG GGAGTCGCTG
     1021 CGACGCTGCC TTCGCCCCGT GCCCCGCTCC GCCGCCGCCT CGCGCCGCCC GCCCCGGCTC
     1081 TGACTGACCG CGTTACTCCC ACAGGTGAGC GGGCGGGACG GCCCTTCTCC TCCGGGCTGT
     1141 AATTAGCTGA GCAAGAGGTA AGGGTTTAAG GGATGGTTGG TTGGTGGGGT ATTAATGTTT
     1201 AATTACCTGG AGCACCTGCC TGAAATCACT TTTTTTCAGG TTGGACCGGT GCCACCATGG
     1261 ACTATAAGGA CCACGACGGA GACTACAAGG ATCATGATAT TGATTACAAA GACGATGACG
     1321 ATAAGATGGC CCCAAAGAAG AAGCGGAAGG TCGGTATCCA CGGAGTCCCA GCAGCCGACA
     1381 AGAAGTACAG CATCGGCCTG GACATCGGCA CCAACTCTGT GGGCTGGGCC GTGATCACCG
     1441 ACGAGTACAA GGTGCCCAGC AAGAAATTCA AGGTGCTGGG CAACACCGAC CGGCACAGCA
     1501 TCAAGAAGAA CCTGATCGGA GCCCTGCTGT TCGACAGCGG CGAAACAGCC GAGGCCACCC
     1561 GGCTGAAGAG AACCGCCAGA AGAAGATACA CCAGACGGAA GAACCGGATC TGCTATCTGC
     1621 AAGAGATCTT CAGCAACGAG ATGGCCAAGG TGGACGACAG CTTCTTCCAC AGACTGGAAG
     1681 AGTCCTTCCT GGTGGAAGAG GATAAGAAGC ACGAGCGGCA CCCCATCTTC GGCAACATCG
     1741 TGGACGAGGT GGCCTACCAC GAGAAGTACC CCACCATCTA CCACCTGAGA AAGAAACTGG
     1801 TGGACAGCAC CGACAAGGCC GACCTGCGGC TGATCTATCT GGCCCTGGCC CACATGATCA
     1861 AGTTCCGGGG CCACTTCCTG ATCGAGGGCG ACCTGAACCC CGACAACAGC GACGTGGACA
     1921 AGCTGTTCAT CCAGCTGGTG CAGACCTACA ACCAGCTGTT CGAGGAAAAC CCCATCAACG
     1981 CCAGCGGCGT GGACGCCAAG GCCATCCTGT CTGCCAGACT GAGCAAGAGC AGACGGCTGG
     2041 AAAATCTGAT CGCCCAGCTG CCCGGCGAGA AGAAGAATGG CCTGTTCGGA AACCTGATTG
     2101 CCCTGAGCCT GGGCCTGACC CCCAACTTCA AGAGCAACTT CGACCTGGCC GAGGATGCCA
     2161 AACTGCAGCT GAGCAAGGAC ACCTACGACG ACGACCTGGA CAACCTGCTG GCCCAGATCG
     2221 GCGACCAGTA CGCCGACCTG TTTCTGGCCG CCAAGAACCT GTCCGACGCC ATCCTGCTGA
     2281 GCGACATCCT GAGAGTGAAC ACCGAGATCA CCAAGGCCCC CCTGAGCGCC TCTATGATCA
     2341 AGAGATACGA CGAGCACCAC CAGGACCTGA CCCTGCTGAA AGCTCTCGTG CGGCAGCAGC
     2401 TGCCTGAGAA GTACAAAGAG ATTTTCTTCG ACCAGAGCAA GAACGGCTAC GCCGGCTACA
     2461 TTGACGGCGG AGCCAGCCAG GAAGAGTTCT ACAAGTTCAT CAAGCCCATC CTGGAAAAGA
     2521 TGGACGGCAC CGAGGAACTG CTCGTGAAGC TGAACAGAGA GGACCTGCTG CGGAAGCAGC
     2581 GGACCTTCGA CAACGGCAGC ATCCCCCACC AGATCCACCT GGGAGAGCTG CACGCCATTC
     2641 TGCGGCGGCA GGAAGATTTT TACCCATTCC TGAAGGACAA CCGGGAAAAG ATCGAGAAGA
     2701 TCCTGACCTT CCGCATCCCC TACTACGTGG GCCCTCTGGC CAGGGGAAAC AGCAGATTCG
     2761 CCTGGATGAC CAGAAAGAGC GAGGAAACCA TCACCCCCTG GAACTTCGAG GAAGTGGTGG
     2821 ACAAGGGCGC TTCCGCCCAG AGCTTCATCG AGCGGATGAC CAACTTCGAT AAGAACCTGC
     2881 CCAACGAGAA GGTGCTGCCC AAGCACAGCC TGCTGTACGA GTACTTCACC GTGTATAACG
     2941 AGCTGACCAA AGTGAAATAC GTGACCGAGG GAATGAGAAA GCCCGCCTTC CTGAGCGGCG
     3001 AGCAGAAAAA GGCCATCGTG GACCTGCTGT TCAAGACCAA CCGGAAAGTG ACCGTGAAGC
     3061 AGCTGAAAGA GGACTACTTC AAGAAAATCG AGTGCTTCGA CTCCGTGGAA ATCTCCGGCG
     3121 TGGAAGATCG GTTCAACGCC TCCCTGGGCA CATACCACGA TCTGCTGAAA ATTATCAAGG
     3181 ACAAGGACTT CCTGGACAAT GAGGAAAACG AGGACATTCT GGAAGATATC GTGCTGACCC
     3241 TGACACTGTT TGAGGACAGA GAGATGATCG AGGAACGGCT GAAAACCTAT GCCCACCTGT
     3301 TCGACGACAA AGTGATGAAG CAGCTGAAGC GGCGGAGATA CACCGGCTGG GGCAGGCTGA
     3361 GCCGGAAGCT GATCAACGGC ATCCGGGACA AGCAGTCCGG CAAGACAATC CTGGATTTCC
     3421 TGAAGTCCGA CGGCTTCGCC AACAGAAACT TCATGCAGCT GATCCACGAC GACAGCCTGA
     3481 CCTTTAAAGA GGACATCCAG AAAGCCCAGG TGTCCGGCCA GGGCGATAGC CTGCACGAGC
     3541 ACATTGCCAA TCTGGCCGGC AGCCCCGCCA TTAAGAAGGG CATCCTGCAG ACAGTGAAGG
     3601 TGGTGGACGA GCTCGTGAAA GTGATGGGCC GGCACAAGCC CGAGAACATC GTGATCGAAA
     3661 TGGCCAGAGA GAACCAGACC ACCCAGAAGG GACAGAAGAA CAGCCGCGAG AGAATGAAGC
     3721 GGATCGAAGA GGGCATCAAA GAGCTGGGCA GCCAGATCCT GAAAGAACAC CCCGTGGAAA
     3781 ACACCCAGCT GCAGAACGAG AAGCTGTACC TGTACTACCT GCAGAATGGG CGGGATATGT
     3841 ACGTGGACCA GGAACTGGAC ATCAACCGGC TGTCCGACTA CGATGTGGAC CATATCGTGC
     3901 CTCAGAGCTT TCTGAAGGAC GACTCCATCG ACAACAAGGT GCTGACCAGA AGCGACAAGA
     3961 ACCGGGGCAA GAGCGACAAC GTGCCCTCCG AAGAGGTCGT GAAGAAGATG AAGAACTACT
     4021 GGCGGCAGCT GCTGAACGCC AAGCTGATTA CCCAGAGAAA GTTCGACAAT CTGACCAAGG
     4081 CCGAGAGAGG CGGCCTGAGC GAACTGGATA AGGCCGGCTT CATCAAGAGA CAGCTGGTGG
     4141 AAACCCGGCA GATCACAAAG CACGTGGCAC AGATCCTGGA CTCCCGGATG AACACTAAGT
     4201 ACGACGAGAA TGACAAGCTG ATCCGGGAAG TGAAAGTGAT CACCCTGAAG TCCAAGCTGG
     4261 TGTCCGATTT CCGGAAGGAT TTCCAGTTTT ACAAAGTGCG CGAGATCAAC AACTACCACC
     4321 ACGCCCACGA CGCCTACCTG AACGCCGTCG TGGGAACCGC CCTGATCAAA AAGTACCCTA
     4381 AGCTGGAAAG CGAGTTCGTG TACGGCGACT ACAAGGTGTA CGACGTGCGG AAGATGATCG
     4441 CCAAGAGCGA GCAGGAAATC GGCAAGGCTA CCGCCAAGTA CTTCTTCTAC AGCAACATCA
     4501 TGAACTTTTT CAAGACCGAG ATTACCCTGG CCAACGGCGA GATCCGGAAG CGGCCTCTGA
     4561 TCGAGACAAA CGGCGAAACC GGGGAGATCG TGTGGGATAA GGGCCGGGAT TTTGCCACCG
     4621 TGCGGAAAGT GCTGAGCATG CCCCAAGTGA ATATCGTGAA AAAGACCGAG GTGCAGACAG
     4681 GCGGCTTCAG CAAAGAGTCT ATCCTGCCCA AGAGGAACAG CGATAAGCTG ATCGCCAGAA
     4741 AGAAGGACTG GGACCCTAAG AAGTACGGCG GCTTCGACAG CCCCACCGTG GCCTATTCTG
     4801 TGCTGGTGGT GGCCAAAGTG GAAAAGGGCA AGTCCAAGAA ACTGAAGAGT GTGAAAGAGC
     4861 TGCTGGGGAT CACCATCATG GAAAGAAGCA GCTTCGAGAA GAATCCCATC GACTTTCTGG
     4921 AAGCCAAGGG CTACAAAGAA GTGAAAAAGG ACCTGATCAT CAAGCTGCCT AAGTACTCCC
     4981 TGTTCGAGCT GGAAAACGGC CGGAAGAGAA TGCTGGCCTC TGCCGGCGAA CTGCAGAAGG
     5041 GAAACGAACT GGCCCTGCCC TCCAAATATG TGAACTTCCT GTACCTGGCC AGCCACTATG
     5101 AGAAGCTGAA GGGCTCCCCC GAGGATAATG AGCAGAAACA GCTGTTTGTG GAACAGCACA
     5161 AGCACTACCT GGACGAGATC ATCGAGCAGA TCAGCGAGTT CTCCAAGAGA GTGATCCTGG
     5221 CCGACGCTAA TCTGGACAAA GTGCTGTCCG CCTACAACAA GCACCGGGAT AAGCCCATCA
     5281 GAGAGCAGGC CGAGAATATC ATCCACCTGT TTACCCTGAC CAATCTGGGA GCCCCTGCCG
     5341 CCTTCAAGTA CTTTGACACC ACCATCGACC GGAAGAGGTA CACCAGCACC AAAGAGGTGC
     5401 TGGACGCCAC CCTGATCCAC CAGAGCATCA CCGGCCTGTA CGAGACACGG ATCGACCTGT
     5461 CTCAGCTGGG AGGCGACAAA AGGCCGGCGG CCACGAAAAA GGCCGGCCAG GCAAAAAAGA
     5521 AAAAGCTTGA GGGCAGAGGA AGTCTGCTAA CATGCGGTGA CGTGGAGGAG AATCCCGGCC
     5581 CTGCTAGCAT GGTGAGCAAG GGCGAGGAGC TGTTCACCGG GGTGGTGCCC ATCCTGGTCG
     5641 AGCTGGACGG CGACGTAAAC GGCCACAAGT TCAGCGTGAG GGGCGAGGGC GAGGGCGATG
     5701 CCACCAACGG CAAGCTGACC CTGAAGTTCA TCTGCACCAC CGGCAAGCTG CCCGTGCCCT
     5761 GGCCCACCCT CGTGACCACC CTGAGCCACG GCGTGCAGTG CTTCGCCCGC TACCCCGACC
     5821 ACATGAAGCA GCACGACTTC TTCAAGTCCG CCATGCCCGA AGGCTACGTC CAGGAGCGCA
     5881 CCATCTTCTT CAAGGACGAC GGCACCTACA AGACCCGCGC CGAGGTGAAG TTCGAGGGCG
     5941 ACACCCTGGT GAACCGCATC GAGCTGAAGG GCGTCGACTT CAAGGAGGAC GGCAACATCC
     6001 TGGGGCACAA GCTGGAGTAC AACTTCAACA GCCACAACAT CTATATCATG GCCGTCAAGC
     6061 AGAAGAACGG CATCAAGGTG AACTTCAAGA TCCGCCACAA CGTGGAGGAC GGCAGCGTGC
     6121 AGCTCGCCGA CCACTACCAG CAGAACACCC CCATCGGCGA CGGCCCCGTG CTGCTGCCCG
     6181 ACAGCCACTA CCTGAGCACC CAGTCCGTGC TGAGCAAAGA CCCCAACGAG AAGCGCGATC
     6241 ACATGGTCCT GCTGGAGTTC CGCACCGCCG CCGGGATCAC TCTCGGCATG GACGAGCTGT
     6301 ACAAGTGAGA ATTCCTAGAG CTCGCTGATC AGCCTCGACT GTGCCTTCTA GTTGCCAGCC
     6361 ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC CTTGACCCTG GAAGGTGCCA CTCCCACTGT
     6421 CCTTTCCTAA TAAAATGAGG AAATTGCATC GCATTGTCTG AGTAGGTGTC ATTCTATTCT
     6481 GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG GGAGGATTGG GAAGAGAATA GCAGGCATGC
     6541 TGGGGAGCGG CCGCAGGAAC CCCTAGTGAT GGAGTTGGCC ACTCCCTCTC TGCGCGCTCG
     6601 CTCGCTCACT GAGGCCGGGC GACCAAAGGT CGCCCGACGC CCGGGCTTTG CCCGGGCGGC
     6661 CTCAGTGAGC GAGCGAGCGC GCAGCTGCCT GCAGGGGCGC CTGATGCGGT ATTTTCTCCT
     6721 TACGCATCTG TGCGGTATTT CACACCGCAT ACGTCAAAGC AACCATAGTA CGCGCCCTGT
     6781 AGCGGCGCAT TAAGCGCGGC GGGTGTGGTG GTTACGCGCA GCGTGACCGC TACACTTGCC
     6841 AGCGCCCTAG CGCCCGCTCC TTTCGCTTTC TTCCCTTCCT TTCTCGCCAC GTTCGCCGGC
     6901 TTTCCCCGTC AAGCTCTAAA TCGGGGGCTC CCTTTAGGGT TCCGATTTAG TGCTTTACGG
     6961 CACCTCGACC CCAAAAAACT TGATTTGGGT GATGGTTCAC GTAGTGGGCC ATCGCCCTGA
     7021 TAGACGGTTT TTCGCCCTTT GACGTTGGAG TCCACGTTCT TTAATAGTGG ACTCTTGTTC
     7081 CAAACTGGAA CAACACTCAA CCCTATCTCG GGCTATTCTT TTGATTTATA AGGGATTTTG
     7141 CCGATTTCGG CCTATTGGTT AAAAAATGAG CTGATTTAAC AAAAATTTAA CGCGAATTTT
     7201 AACAAAATAT TAACGTTTAC AATTTTATGG TGCACTCTCA GTACAATCTG CTCTGATGCC
     7261 GCATAGTTAA GCCAGCCCCG ACACCCGCCA ACACCCGCTG ACGCGCCCTG ACGGGCTTGT
     7321 CTGCTCCCGG CATCCGCTTA CAGACAAGCT GTGACCGTCT CCGGGAGCTG CATGTGTCAG
     7381 AGGTTTTCAC CGTCATCACC GAAACGCGCG AGACGAAAGG GCCTCGTGAT ACGCCTATTT
     7441 TTATAGGTTA ATGTCATGAT AATAATGGTT TCTTAGACGT CAGGTGGCAC TTTTCGGGGA
     7501 AATGTGCGCG GAACCCCTAT TTGTTTATTT TTCTAAATAC ATTCAAATAT GTATCCGCTC
     7561 ATGAGACAAT AACCCTGATA AATGCTTCAA TAATATTGAA AAAGGAAGAG TATGAGTATT
     7621 CAACATTTCC GTGTCGCCCT TATTCCCTTT TTTGCGGCAT TTTGCCTTCC TGTTTTTGCT
     7681 CACCCAGAAA CGCTGGTGAA AGTAAAAGAT GCTGAAGATC AGTTGGGTGC ACGAGTGGGT
     7741 TACATCGAAC TGGATCTCAA CAGCGGTAAG ATCCTTGAGA GTTTTCGCCC CGAAGAACGT
     7801 TTTCCAATGA TGAGCACTTT TAAAGTTCTG CTATGTGGCG CGGTATTATC CCGTATTGAC
     7861 GCCGGGCAAG AGCAACTCGG TCGCCGCATA CACTATTCTC AGAATGACTT GGTTGAGTAC
     7921 TCACCAGTCA CAGAAAAGCA TCTTACGGAT GGCATGACAG TAAGAGAATT ATGCAGTGCT
     7981 GCCATAACCA TGAGTGATAA CACTGCGGCC AACTTACTTC TGACAACGAT CGGAGGACCG
     8041 AAGGAGCTAA CCGCTTTTTT GCACAACATG GGGGATCATG TAACTCGCCT TGATCGTTGG
     8101 GAACCGGAGC TGAATGAAGC CATACCAAAC GACGAGCGTG ACACCACGAT GCCTGTAGCA
     8161 ATGGCAACAA CGTTGCGCAA ACTATTAACT GGCGAACTAC TTACTCTAGC TTCCCGGCAA
     8221 CAATTAATAG ACTGGATGGA GGCGGATAAA GTTGCAGGAC CACTTCTGCG CTCGGCCCTT
     8281 CCGGCTGGCT GGTTTATTGC TGATAAATCT GGAGCCGGTG AGCGTGGAAG CCGCGGTATC
     8341 ATTGCAGCAC TGGGGCCAGA TGGTAAGCCC TCCCGTATCG TAGTTATCTA CACGACGGGG
     8401 AGTCAGGCAA CTATGGATGA ACGAAATAGA CAGATCGCTG AGATAGGTGC CTCACTGATT
     8461 AAGCATTGGT AACTGTCAGA CCAAGTTTAC TCATATATAC TTTAGATTGA TTTAAAACTT
     8521 CATTTTTAAT TTAAAAGGAT CTAGGTGAAG ATCCTTTTTG ATAATCTCAT GACCAAAATC
     8581 CCTTAACGTG AGTTTTCGTT CCACTGAGCG TCAGACCCCG TAGAAAAGAT CAAAGGATCT
     8641 TCTTGAGATC CTTTTTTTCT GCGCGTAATC TGCTGCTTGC AAACAAAAAA ACCACCGCTA
     8701 CCAGCGGTGG TTTGTTTGCC GGATCAAGAG CTACCAACTC TTTTTCCGAA GGTAACTGGC
     8761 TTCAGCAGAG CGCAGATACC AAATACTGTC CTTCTAGTGT AGCCGTAGTT AGGCCACCAC
     8821 TTCAAGAACT CTGTAGCACC GCCTACATAC CTCGCTCTGC TAATCCTGTT ACCAGTGGCT
     8881 GCTGCCAGTG GCGATAAGTC GTGTCTTACC GGGTTGGACT CAAGACGATA GTTACCGGAT
     8941 AAGGCGCAGC GGTCGGGCTG AACGGGGGGT TCGTGCACAC AGCCCAGCTT GGAGCGAACG
     9001 ACCTACACCG AACTGAGATA CCTACAGCGT GAGCTATGAG AAAGCGCCAC GCTTCCCGAA
     9061 GGGAGAAAGG CGGACAGGTA TCCGGTAAGC GGCAGGGTCG GAACAGGAGA GCGCACGAGG
     9121 GAGCTTCCAG GGGGAAACGC CTGGTATCTT TATAGTCCTG TCGGGTTTCG CCACCTCTGA
     9181 CTTGAGCGTC GATTTTTGTG ATGCTCGTCA GGGGGGCGGA GCCTATGGAA AAACGCCAGC
     9241 AACGCGGCCT TTTTACGGTT CCTGGCCTTT TGCTGGCCTT TTGCTCACAT GT
//
