LOCUS       PX260a                 10223 bp    DNA     circular     12-FEB-2013
FEATURES             Location/Qualifiers
     misc_feature    1321..5421
                     /note="hSpCsn1"
     misc_feature    5479..5710
                     /note="bGH polyA"
     rep_origin      7718..8024
                     /dnas_title="f1 Origin"
                     /vntifkey="33"
                     /label=f1 Origin
     misc_feature    8543..9400
                     /dnas_title="AmpR"
                     /vntifkey="21"
                     /label=AmpR
     misc_feature    9258..9263
                     /note="BsaI"
     rep_origin      9551..10218
                     /dnas_title="pUC ori"
                     /vntifkey="33"
                     /label=pUC ori
     misc_feature    1345..1347
                     /note="D10"
     misc_feature    6235..6826
                     /note="PGK"
     misc_feature    6829..7428
                     /note="Puro"
     promoter        1..249
                     /dnas_title="Human U6 Promoter"
                     /vntifkey="29"
                     /label=Human U6 Promoter
     misc_feature    5719..5945
                     /note="H1 Promoter"
     misc_feature    5946..6032
                     /note="tracrRNA"
     misc_feature    390..1188
                     /note="CBh"
     misc_feature    5422..5469
                     /note="NLS"
     misc_feature    252..287
                     /note="DR"
     misc_feature    304..339
                     /note="DR"
     misc_feature    340..344
                     /note="U6 terminator"
     source          1..10223
                     /dnas_title="PX260a -
                     U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro"
     misc_feature    1201..1269
                     /note="3xFLAG"
     misc_feature    1270..1320
                     /note="NLS"
ORIGIN      
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt GTGGAAAGGA
      241 CGAAACACCG GGTTTTAGAG CTATGCTGTT TTGAATGGTC CCAAAACggG TCTTCgaGAA
      301 GACGTTTTAG AGCTATGCTG TTTTGAATGG TCCCAAAACT TTTTctagcg cgtgcgccaa
      361 ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct
      421 ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc
      481 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      541 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      601 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      661 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      721 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      781 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg
      841 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
      901 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
      961 gctgcgacgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     1021 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     1081 ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat
     1141 gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc
     1201 ATGGACTATA AGGACCACGA CGGAGACTAC AAGGATCATG ATATTGATTA CAAAGACGAT
     1261 GACGATAAGA TGGCCCCAAA GAAGAAGCGG AAGGTCGGTA TCCACGGAGT CCCAGCAGCC
     1321 GACAAGAAGT ACAGCATCGG CCTGGACATC GGCACCAACT CTGTGGGCTG GGCCGTGATC
     1381 ACCGACGAGT ACAAGGTGCC CAGCAAGAAA TTCAAGGTGC TGGGCAACAC CGACCGGCAC
     1441 AGCATCAAGA AGAACCTGAT CGGAGCCCTG CTGTTCGACA GCGGCGAAAC AGCCGAGGCC
     1501 ACCCGGCTGA AGAGAACCGC CAGAAGAAGA TACACCAGAC GGAAGAACCG GATCTGCTAT
     1561 CTGCAAGAGA TCTTCAGCAA CGAGATGGCC AAGGTGGACG ACAGCTTCTT CCACAGACTG
     1621 GAAGAGTCCT TCCTGGTGGA AGAGGATAAG AAGCACGAGC GGCACCCCAT CTTCGGCAAC
     1681 ATCGTGGACG AGGTGGCCTA CCACGAGAAG TACCCCACCA TCTACCACCT GAGAAAGAAA
     1741 CTGGTGGACA GCACCGACAA GGCCGACCTG CGGCTGATCT ATCTGGCCCT GGCCCACATG
     1801 ATCAAGTTCC GGGGCCACTT CCTGATCGAG GGCGACCTGA ACCCCGACAA CAGCGACGTG
     1861 GACAAGCTGT TCATCCAGCT GGTGCAGACC TACAACCAGC TGTTCGAGGA AAACCCCATC
     1921 AACGCCAGCG GCGTGGACGC CAAGGCCATC CTGTCTGCCA GACTGAGCAA GAGCAGACGG
     1981 CTGGAAAATC TGATCGCCCA GCTGCCCGGC GAGAAGAAGA ATGGCCTGTT CGGAAACCTG
     2041 ATTGCCCTGA GCCTGGGCCT GACCCCCAAC TTCAAGAGCA ACTTCGACCT GGCCGAGGAT
     2101 GCCAAACTGC AGCTGAGCAA GGACACCTAC GACGACGACC TGGACAACCT GCTGGCCCAG
     2161 ATCGGCGACC AGTACGCCGA CCTGTTTCTG GCCGCCAAGA ACCTGTCCGA CGCCATCCTG
     2221 CTGAGCGACA TCCTGAGAGT GAACACCGAG ATCACCAAGG CCCCCCTGAG CGCCTCTATG
     2281 ATCAAGAGAT ACGACGAGCA CCACCAGGAC CTGACCCTGC TGAAAGCTCT CGTGCGGCAG
     2341 CAGCTGCCTG AGAAGTACAA AGAGATTTTC TTCGACCAGA GCAAGAACGG CTACGCCGGC
     2401 TACATTGACG GCGGAGCCAG CCAGGAAGAG TTCTACAAGT TCATCAAGCC CATCCTGGAA
     2461 AAGATGGACG GCACCGAGGA ACTGCTCGTG AAGCTGAACA GAGAGGACCT GCTGCGGAAG
     2521 CAGCGGACCT TCGACAACGG CAGCATCCCC CACCAGATCC ACCTGGGAGA GCTGCACGCC
     2581 ATTCTGCGGC GGCAGGAAGA TTTTTACCCA TTCCTGAAGG ACAACCGGGA AAAGATCGAG
     2641 AAGATCCTGA CCTTCCGCAT CCCCTACTAC GTGGGCCCTC TGGCCAGGGG AAACAGCAGA
     2701 TTCGCCTGGA TGACCAGAAA GAGCGAGGAA ACCATCACCC CCTGGAACTT CGAGGAAGTG
     2761 GTGGACAAGG GCGCTTCCGC CCAGAGCTTC ATCGAGCGGA TGACCAACTT CGATAAGAAC
     2821 CTGCCCAACG AGAAGGTGCT GCCCAAGCAC AGCCTGCTGT ACGAGTACTT CACCGTGTAT
     2881 AACGAGCTGA CCAAAGTGAA ATACGTGACC GAGGGAATGA GAAAGCCCGC CTTCCTGAGC
     2941 GGCGAGCAGA AAAAGGCCAT CGTGGACCTG CTGTTCAAGA CCAACCGGAA AGTGACCGTG
     3001 AAGCAGCTGA AAGAGGACTA CTTCAAGAAA ATCGAGTGCT TCGACTCCGT GGAAATCTCC
     3061 GGCGTGGAAG ATCGGTTCAA CGCCTCCCTG GGCACATACC ACGATCTGCT GAAAATTATC
     3121 AAGGACAAGG ACTTCCTGGA CAATGAGGAA AACGAGGACA TTCTGGAAGA TATCGTGCTG
     3181 ACCCTGACAC TGTTTGAGGA CAGAGAGATG ATCGAGGAAC GGCTGAAAAC CTATGCCCAC
     3241 CTGTTCGACG ACAAAGTGAT GAAGCAGCTG AAGCGGCGGA GATACACCGG CTGGGGCAGG
     3301 CTGAGCCGGA AGCTGATCAA CGGCATCCGG GACAAGCAGT CCGGCAAGAC AATCCTGGAT
     3361 TTCCTGAAGT CCGACGGCTT CGCCAACAGA AACTTCATGC AGCTGATCCA CGACGACAGC
     3421 CTGACCTTTA AAGAGGACAT CCAGAAAGCC CAGGTGTCCG GCCAGGGCGA TAGCCTGCAC
     3481 GAGCACATTG CCAATCTGGC CGGCAGCCCC GCCATTAAGA AGGGCATCCT GCAGACAGTG
     3541 AAGGTGGTGG ACGAGCTCGT GAAAGTGATG GGCCGGCACA AGCCCGAGAA CATCGTGATC
     3601 GAAATGGCCA GAGAGAACCA GACCACCCAG AAGGGACAGA AGAACAGCCG CGAGAGAATG
     3661 AAGCGGATCG AAGAGGGCAT CAAAGAGCTG GGCAGCCAGA TCCTGAAAGA ACACCCCGTG
     3721 GAAAACACCC AGCTGCAGAA CGAGAAGCTG TACCTGTACT ACCTGCAGAA TGGGCGGGAT
     3781 ATGTACGTGG ACCAGGAACT GGACATCAAC CGGCTGTCCG ACTACGATGT GGACCATATC
     3841 GTGCCTCAGA GCTTTCTGAA GGACGACTCC ATCGACAACA AGGTGCTGAC CAGAAGCGAC
     3901 AAGAACCGGG GCAAGAGCGA CAACGTGCCC TCCGAAGAGG TCGTGAAGAA GATGAAGAAC
     3961 TACTGGCGGC AGCTGCTGAA CGCCAAGCTG ATTACCCAGA GAAAGTTCGA CAATCTGACC
     4021 AAGGCCGAGA GAGGCGGCCT GAGCGAACTG GATAAGGCCG GCTTCATCAA GAGACAGCTG
     4081 GTGGAAACCC GGCAGATCAC AAAGCACGTG GCACAGATCC TGGACTCCCG GATGAACACT
     4141 AAGTACGACG AGAATGACAA GCTGATCCGG GAAGTGAAAG TGATCACCCT GAAGTCCAAG
     4201 CTGGTGTCCG ATTTCCGGAA GGATTTCCAG TTTTACAAAG TGCGCGAGAT CAACAACTAC
     4261 CACCACGCCC ACGACGCCTA CCTGAACGCC GTCGTGGGAA CCGCCCTGAT CAAAAAGTAC
     4321 CCTAAGCTGG AAAGCGAGTT CGTGTACGGC GACTACAAGG TGTACGACGT GCGGAAGATG
     4381 ATCGCCAAGA GCGAGCAGGA AATCGGCAAG GCTACCGCCA AGTACTTCTT CTACAGCAAC
     4441 ATCATGAACT TTTTCAAGAC CGAGATTACC CTGGCCAACG GCGAGATCCG GAAGCGGCCT
     4501 CTGATCGAGA CAAACGGCGA AACCGGGGAG ATCGTGTGGG ATAAGGGCCG GGATTTTGCC
     4561 ACCGTGCGGA AAGTGCTGAG CATGCCCCAA GTGAATATCG TGAAAAAGAC CGAGGTGCAG
     4621 ACAGGCGGCT TCAGCAAAGA GTCTATCCTG CCCAAGAGGA ACAGCGATAA GCTGATCGCC
     4681 AGAAAGAAGG ACTGGGACCC TAAGAAGTAC GGCGGCTTCG ACAGCCCCAC CGTGGCCTAT
     4741 TCTGTGCTGG TGGTGGCCAA AGTGGAAAAG GGCAAGTCCA AGAAACTGAA GAGTGTGAAA
     4801 GAGCTGCTGG GGATCACCAT CATGGAAAGA AGCAGCTTCG AGAAGAATCC CATCGACTTT
     4861 CTGGAAGCCA AGGGCTACAA AGAAGTGAAA AAGGACCTGA TCATCAAGCT GCCTAAGTAC
     4921 TCCCTGTTCG AGCTGGAAAA CGGCCGGAAG AGAATGCTGG CCTCTGCCGG CGAACTGCAG
     4981 AAGGGAAACG AACTGGCCCT GCCCTCCAAA TATGTGAACT TCCTGTACCT GGCCAGCCAC
     5041 TATGAGAAGC TGAAGGGCTC CCCCGAGGAT AATGAGCAGA AACAGCTGTT TGTGGAACAG
     5101 CACAAGCACT ACCTGGACGA GATCATCGAG CAGATCAGCG AGTTCTCCAA GAGAGTGATC
     5161 CTGGCCGACG CTAATCTGGA CAAAGTGCTG TCCGCCTACA ACAAGCACCG GGATAAGCCC
     5221 ATCAGAGAGC AGGCCGAGAA TATCATCCAC CTGTTTACCC TGACCAATCT GGGAGCCCCT
     5281 GCCGCCTTCA AGTACTTTGA CACCACCATC GACCGGAAGA GGTACACCAG CACCAAAGAG
     5341 GTGCTGGACG CCACCCTGAT CCACCAGAGC ATCACCGGCC TGTACGAGAC ACGGATCGAC
     5401 CTGTCTCAGC TGGGAGGCGA CAAAAGGCCG GCGGCCACGA AAAAGGCCGG CCAGGCAAAA
     5461 AAGAAAAAGt aagaattcCT AGAGCTCGCT GATCAGCCTC GACTGTGCCT TCTAGTTGCC
     5521 AGCCATCTGT TGTTTGCCCC TCCCCCGTGC CTTCCTTGAC CCTGGAAGGT GCCACTCCCA
     5581 CTGTCCTTTC CTAATAAAAT GAGGAAATTG CATCGCATTG TCTGAGTAGG TGTCATTCTA
     5641 TTCTGGGGGG TGGGGTGGGG CAGGACAGCA AGGGGGAGGA TTGGGAAGAg AATAGCAGGC
     5701 ATGCTGGGGA gcggccgcAA TTCGAACGCT GACGTCATCA ACCCGCTCCA AGGAATCGCG
     5761 GGCCCAGTGT CACTAGGCGG GAACACCCAG CGCGCGTGCG CCCTGGCAGG AAGATGGCTG
     5821 TGAGGGACAG GGGAGTGGCG CCCTGCAATA TTTGCATGTC GCTATGTGTT CTGGGAAATC
     5881 ACCATAAACG TGAAATGTCT TTGGATTTGG GAATCTTATA AGTTCTGTAT GAGACCACAG
     5941 ATCTAgttgg aaccattcaa aacagcatag caagttaaaa taaggctagt ccgttatcaa
     6001 cttgaaaaag tggcaccgag tcggtgcttt ttggcgccga caaccactac ctgagcaccc
     6061 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     6121 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaGAA TTCcgatcat
     6181 attcaataac ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtctga
     6241 agaggagttt acgtccagcc aagcttagga tctcgacctc gaaattctac cgggtagggg
     6301 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
     6361 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagc gccaaccggc
     6421 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     6481 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     6541 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     6601 ccaatagcag ctttgctcct tcgctttctg ggctcagcag ctgggaaggg tgggtccggg
     6661 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
     6721 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
     6781 ggcctttcga cctgcatcca tctagatctc gagcagctga agcttaccat gaccgagtac
     6841 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     6901 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     6961 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     7021 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     7081 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     7141 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     7201 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     7261 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     7321 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     7381 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     7441 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgatgat atcagatccc
     7501 cgggatgcag aaattgatga tctattaaac aataaagatg tccactaaaa tggaagtttt
     7561 tcctgtcata ctttgttaag aagggtgaga acagagtacc tacattttga atggaaggat
     7621 tggagcggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     7681 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     7741 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     7801 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     7861 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     7921 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     7981 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     8041 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     8101 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     8161 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     8221 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     8281 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     8341 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     8401 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     8461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     8521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     8581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     8641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     8701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     8761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     8821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     8881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     8941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     9001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     9061 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     9121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     9181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     9241 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     9301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     9361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     9421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     9481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     9541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     9601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     9661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     9721 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     9781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     9841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     9901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     9961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
    10021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
    10081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
    10141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
    10201 ttgctggcct tttgctcaca tgt
//

