LOCUS Exported 10000 bp ds-DNA circular SYN 30-SEP-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS PX330_ER50_cas9_ER50 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10000) AUTHORS . TITLE Direct Submission JOURNAL Exported Friday, Sep 30, 2016 from SnapGene 3.2.1 http://www.snapgene.com COMMENT LOCUS PX330 8506 bp DNA circular 12-FEB-2013 COMMENT VNTAUTHOR|Le Cong| COMMENT VNTAUTHOREML|congle.thu@gmail.com| FEATURES Location/Qualifiers source 1..10000 /organism="synthetic DNA construct" /note="PX330a-U6-chimeric_BB" promoter 1..249 /note="Human U6 Promoter" misc_feature 268..343 /note="chimeric guide RNA scaffold" misc_feature 344..349 /note="U6 terminator" misc_feature 440..1238 /note="CBh" misc_feature 1239..6230 /note="hSpCsn1" misc_feature 1320..1370 /note="NLS" misc_feature 1377..2111 /note="ER50" misc_feature 2130..6230 /note="hSpCas9" misc_feature 6231..6278 /note="NLS" misc_feature 6231..6278 /note="NLS" misc_feature 6279..7013 /note="ER50" misc_feature 7023..7254 /note="bGH polyA" misc_feature 7263..7403 /note="R-ITR" rep_origin 7495..7801 /direction=RIGHT /note="f1 Origin" misc_feature 8320..9177 /note="AmpR" rep_origin 9328..9995 /direction=RIGHT /note="pUC ori" 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 ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag 301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct 361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac 421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc 541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt 661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc 1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg 1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag 1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac 1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata 1261 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga 1321 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc ggatccagcc 1381 ttgccctgtc acttacagcc gaccagatgg tttccgcgct tctcgacgct gaacctccaa 1441 ttctctattc cgaatacgac ccaaccaggc cgttctccga ggcatctatg atgggtctgc 1501 tgacaaatct ggcagacagg gaactggtgc acatgatcaa ttgggcgaag cgcgtacccg 1561 gattcgtcga tcttgcactc catgatcagg tgcacttgct ggagtgcgct tggatggaga 1621 tcctcatgat cgggctggtg tggcggagta tggaacaccc cggcaagttg ctgtttgcgc 1681 ctaacctcct gttggacagg aaccagggga aatgtgtgga gggcggtgtg gaaatctttg 1741 acatgctcct cgctacctca agccggttta ggatgatgaa tctgcagggc gaagagttcg 1801 tgtgtctcaa atctatcata ctgttgaaca gcggagtcta caccttcctc tccagtactc 1861 tgaaatctct ggaggagaaa gatcatatcc atcgcgtgct ggacaagata accgacacgt 1921 tgattcactt gatggccaaa gctgggctca ctctgcaaca acaacatcag cgactggcac 1981 agctgttgct gattttgagc cacattcggc acatgtccag caagagaatg gagcacctct 2041 atagtatgaa gtgcaagaac gtcgtacccc tgtcagatct gcttcttgaa atgcttgatg 2101 cccaccggct gggaagcgga agcggatccg acaagaagta cagcatcggc ctggacatcg 2161 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat 2221 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc 2281 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat 2341 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca 2401 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga 2461 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt 2521 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc 2581 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg 2641 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct 2701 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc 2761 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg 2821 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact 2881 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg 2941 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg 3001 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga 3061 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc 3121 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct 3181 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt 3241 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga 3301 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc 3361 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat 3421 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg 3481 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa 3541 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca 3601 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca 3661 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg 3721 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc 3781 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa 3841 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg 3901 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa 3961 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga 4021 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga 4081 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg 4141 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa 4201 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc 4261 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg 4321 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg 4381 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga 4441 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg 4501 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt 4561 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc 4621 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca 4681 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct 4741 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga 4801 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg 4861 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg 4921 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg 4981 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt 5041 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg 5101 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg 5161 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg 5221 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc 5281 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga 5341 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag 5401 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc 5461 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg 5521 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg 5581 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa 5641 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa 5701 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga 5761 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat 5821 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata 5881 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc 5941 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt 6001 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc 6061 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg 6121 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca 6181 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg 6241 cggccacgaa aaaggccggc caggcaaaaa agaaaaagag ccttgccctg tcacttacag 6301 ccgaccagat ggtttccgcg cttctcgacg ctgaacctcc aattctctat tccgaatacg 6361 acccaaccag gccgttctcc gaggcatcta tgatgggtct gctgacaaat ctggcagaca 6421 gggaactggt gcacatgatc aattgggcga agcgcgtacc cggattcgtc gatcttgcac 6481 tccatgatca ggtgcacttg ctggagtgcg cttggatgga gatcctcatg atcgggctgg 6541 tgtggcggag tatggaacac cccggcaagt tgctgtttgc gcctaacctc ctgttggaca 6601 ggaaccaggg gaaatgtgtg gagggcggtg tggaaatctt tgacatgctc ctcgctacct 6661 caagccggtt taggatgatg aatctgcagg gcgaagagtt cgtgtgtctc aaatctatca 6721 tactgttgaa cagcggagtc tacaccttcc tctccagtac tctgaaatct ctggaggaga 6781 aagatcatat ccatcgcgtg ctggacaaga taaccgacac gttgattcac ttgatggcca 6841 aagctgggct cactctgcaa caacaacatc agcgactggc acagctgttg ctgattttga 6901 gccacattcg gcacatgtcc agcaagagaa tggagcacct ctatagtatg aagtgcaaga 6961 acgtcgtacc cctgtcagat ctgcttcttg aaatgcttga tgcccaccgg ctgtaagaat 7021 tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 7081 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 7141 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 7201 ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc 7261 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 7321 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga 7381 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg 7441 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta 7501 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 7561 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 7621 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 7681 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt 7741 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 7801 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc 7861 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta 7921 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 7981 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 8041 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 8101 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 8161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 8221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 8281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 8341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 8401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 8461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 8521 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 8581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 8641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 8701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 8761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 8821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 8881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 8941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 9001 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg 9061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 9121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 9181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 9241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 9301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 9361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 9421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 9481 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct 9541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 9601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 9661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 9721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 9781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 9841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 9901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 9961 ttacggttcc tggccttttg ctggcctttt gctcacatgt //