LOCUS AAV_SaCas9_Backb 7216 bp DNA circular 26-JUN-2013 DEFINITION . ACCESSION urn.local.zhangf.1372221258010.8 VERSION KEYWORDS . SOURCE null ORGANISM . FEATURES Location/Qualifiers misc_feature 1..130 /label=L-ITR misc_feature 137..142 /note="XbaI" /note="Geneious type: restriction" /label=XbaI misc_feature 143..148 /note="XhoI" /label=XhoI promoter 149..749 /note="CMV" /label=CMV Promoter misc_feature 4113..4344 /note="bGH polyA" /label=bGH polyA misc_feature 4345..4350 /note="KpnI" /label=KpnI misc_feature 4621..4701 /label=SaCas9 sgRNA scaffold misc_feature 4710..4850 /label=R-ITR rep_origin 4942..5248 /label=f1 Origin misc_feature 5767..6624 /label=AmpR rep_origin 6775..7442 /label=pUC ori promoter 4352..4600 /dnas_title="Human U6 Promoter" /vntifkey="29" /label=Human U6 Promoter misc_feature 762..812 /note="NLS" exon 813..3968 /note="hSaCas9" misc_feature 1212..1214 /note="Ub high" misc_feature 3015..3017 /note="Ub high - AAG = K (original), AGG = R" misc_feature 3969..4016 /note="NLS" misc_feature 4017..4022 /note="BamHI" /label=BamHI CDS 4023..4106 /label=3xHA /note="Geneious type: restriction" /label=EcoRI misc_feature 4702..4709 /note="NotI" misc_feature 4601..4620 /note="BsaI (GGTCTCN1,N5)" source 1..7447 /dnas_title="CMV-SaCas9(reopt)-3xHA-U6-BsaI(sgRNA) (V2" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggcctcta gactcgaggc gttgacattg attattgact agttattaat 181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 541 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 661 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 721 tctatataag cagagctctc tggctaacta ccggtgccac cATGGCCCCA AAGAAGAAGC 781 GGAAGGTCGG TATCCACGGA GTCCCAGCAG CCAAGCGGAA CTACATCCTG GGCCTGGACA 841 TCGGCATCAC CAGCGTGGGC TACGGCATCA TCGACTACGA GACACGGGAC GTGATCGATG 901 CCGGCGTGCG GCTGTTCAAA GAGGCCAACG TGGAAAACAA CGAGGGCAGG CGGAGCAAGA 961 GAGGCGCCAG AAGGCTGAAG CGGCGGAGGC GGCATAGAAT CCAGAGAGTG AAGAAGCTGC 1021 TGTTCGACTA CAACCTGCTG ACCGACCACA GCGAGCTGAG CGGCATCAAC CCCTACGAGG 1081 CCAGAGTGAA GGGCCTGAGC CAGAAGCTGA GCGAGGAAGA GTTCTCTGCC GCCCTGCTGC 1141 ACCTGGCCAA GAGAAGAGGC GTGCACAACG TGAACGAGGT GGAAGAGGAC ACCGGCAACG 1201 AGCTGTCCAC CAAAGAGCAG ATCAGCCGGA ACAGCAAGGC CCTGGAAGAG AAATACGTGG 1261 CCGAACTGCA GCTGGAACGG CTGAAGAAAG ACGGCGAAGT GCGGGGCAGC ATCAACAGAT 1321 TCAAGACCAG CGACTACGTG AAAGAAGCCA AACAGCTGCT GAAGGTGCAG AAGGCCTACC 1381 ACCAGCTGGA CCAGAGCTTC ATCGACACCT ACATCGACCT GCTGGAAACC CGGCGGACCT 1441 ACTATGAGGG ACCTGGCGAG GGCAGCCCCT TCGGCTGGAA GGACATCAAA GAATGGTACG 1501 AGATGCTGAT GGGCCACTGC ACCTACTTCC CCGAGGAACT GCGGAGCGTG AAGTACGCCT 1561 ACAACGCCGA CCTGTACAAC GCCCTGAACG ACCTGAACAA TCTCGTGATC ACCAGGGACG 1621 AGAACGAGAA GCTGGAATAT TACGAGAAGT TCCAGATCAT CGAGAACGTG TTCAAGCAGA 1681 AGAAGAAGCC CACCCTGAAG CAGATCGCCA AAGAAATCCT CGTGAACGAA GAGGATATTA 1741 AGGGCTACAG AGTGACCAGC ACCGGCAAGC CCGAGTTCAC CAACCTGAAG GTGTACCACG 1801 ACATCAAGGA CATTACCGCC CGGAAAGAGA TTATTGAGAA CGCCGAGCTG CTGGATCAGA 1861 TTGCCAAGAT CCTGACCATC TACCAGAGCA GCGAGGACAT CCAGGAAGAA CTGACCAATC 1921 TGAACTCCGA GCTGACCCAG GAAGAGATCG AGCAGATCTC TAATCTGAAG GGCTATACCG 1981 GCACCCACAA CCTGAGCCTG AAGGCCATCA ACCTGATCCT GGACGAGCTG TGGCACACCA 2041 ACGACAACCA GATCGCTATC TTCAACCGGC TGAAGCTGGT GCCCAAGAAG GTGGACCTGT 2101 CCCAGCAGAA AGAGATCCCC ACCACCCTGG TGGACGACTT CATCCTGAGC CCCGTCGTGA 2161 AGAGAAGCTT CATCCAGAGC ATCAAAGTGA TCAACGCCAT CATCAAGAAG TACGGCCTGC 2221 CCAACGACAT CATTATCGAG CTGGCCCGCG AGAAGAACTC CAAGGACGCC CAGAAAATGA 2281 TCAACGAGAT GCAGAAGCGG AACCGGCAGA CCAACGAGCG GATCGAGGAA ATCATCCGGA 2341 CCACCGGCAA AGAGAACGCC AAGTACCTGA TCGAGAAGAT CAAGCTGCAC GACATGCAGG 2401 AAGGCAAGTG CCTGTACAGC CTGGAAGCCA TCCCTCTGGA AGATCTGCTG AACAACCCCT 2461 TCAACTATGA GGTGGACCAC ATCATCCCCA GAAGCGTGTC CTTCGACAAC AGCTTCAACA 2521 ACAAGGTGCT CGTGAAGCAG GAAGAAAACA GCAAGAAGGG CAACCGGACC CCATTCCAGT 2581 ACCTGAGCAG CAGCGACAGC AAGATCAGCT ACGAAACCTT CAAGAAGCAC ATCCTGAATC 2641 TGGCCAAGGG CAAGGGCAGA ATCAGCAAGA CCAAGAAAGA GTATCTGCTG GAAGAACGGG 2701 ACATCAACAG GTTCTCCGTG CAGAAAGACT TCATCAACCG GAACCTGGTG GATACCAGAT 2761 ACGCCACCAG AGGCCTGATG AACCTGCTGC GGAGCTACTT CAGAGTGAAC AACCTGGACG 2821 TGAAAGTGAA GTCCATCAAT GGCGGCTTCA CCAGCTTTCT GCGGCGGAAG TGGAAGTTTA 2881 AGAAAGAGCG GAACAAGGGG TACAAGCACC ACGCCGAGGA CGCCCTGATC ATTGCCAACG 2941 CCGATTTCAT CTTCAAAGAG TGGAAGAAAC TGGACAAGGC CAAAAAAGTG ATGGAAAACC 3001 AGATGTTCGA GGAAAAGCAG GCCGAGAGCA TGCCCGAGAT CGAAACCGAG CAGGAGTACA 3061 AAGAGATCTT CATCACCCCC CACCAGATCA AGCACATTAA GGACTTCAAG GACTACAAGT 3121 ACAGCCACCG GGTGGACAAG AAGCCTAATA GAGAGCTGAT TAACGACACC CTGTACTCCA 3181 CCCGGAAGGA CGACAAGGGC AACACCCTGA TCGTGAACAA TCTGAACGGC CTGTACGACA 3241 AGGACAATGA CAAGCTGAAA AAGCTGATCA ACAAGAGCCC CGAAAAGCTG CTGATGTACC 3301 ACCACGACCC CCAGACCTAC CAGAAACTGA AGCTGATTAT GGAACAGTAC GGCGACGAGA 3361 AGAATCCCCT GTACAAGTAC TACGAGGAAA CCGGGAACTA CCTGACCAAG TACTCCAAAA 3421 AGGACAACGG CCCCGTGATC AAGAAGATTA AGTATTACGG CAACAAACTG AACGCCCATC 3481 TGGACATCAC CGACGACTAC CCCAACAGCA GAAACAAGGT CGTGAAGCTG TCCCTGAAGC 3541 CCTACAGATT CGACGTGTAC CTGGACAATG GCGTGTACAA GTTCGTGACC GTGAAGAATC 3601 TGGATGTGAT CAAAAAAGAA AACTACTACG AAGTGAATAG CAAGTGCTAT GAGGAAGCTA 3661 AGAAGCTGAA GAAGATCAGC AACCAGGCCG AGTTTATCGC CTCCTTCTAC AACAACGATC 3721 TGATCAAGAT CAACGGCGAG CTGTATAGAG TGATCGGCGT GAACAACGAC CTGCTGAACC 3781 GGATCGAAGT GAACATGATC GACATCACCT ACCGCGAGTA CCTGGAAAAC ATGAACGACA 3841 AGAGGCCCCC CAGGATCATT AAGACAATCG CCTCCAAGAC CCAGAGCATT AAGAAGTACA 3901 GCACAGACAT TCTGGGCAAC CTGTATGAAG TGAAATCTAA GAAGCACCCT CAGATCATCA 3961 AAAAGGGCAA AAGGCCGGCG GCCACGAAAA AGGCCGGCCA GGCAAAAAAG AAAAAGggat 4021 cctacccata cgatgttcca gattacgctt acccatacga tgttccagat tacgcttacc 4081 catacgatgt tccagattac gcttaagaat tcctagagct cgctgatcag cctcgactgt 4141 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 4201 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 4261 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 4321 agagaatagc aggcatgctg gggaggtacc tgagggccta tttcccatga ttccttcata 4381 tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac 4441 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt 4501 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga 4561 tttcttggct ttatatatct tGTGGAAAGG ACGAAACACC gGAGACCacg gcaGGTCTCa 4621 gttttagtac tctggaaaca gaatctacta aaacaaggca aaatgccgtg tttatctcgt 4681 caacttgttg gcgagatttt tgcggccgca ggaaccccta gtgatggagt tggccactcc 4741 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg 4801 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat 4861 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca 4921 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 4981 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 5041 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 5101 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt 5161 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 5221 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat 5281 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 5341 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca 5401 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 5461 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 5521 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 5581 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 5641 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 5701 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 5761 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 5821 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 5881 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 5941 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 6001 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 6061 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 6121 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 6181 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 6241 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 6301 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 6361 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 6421 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 6481 ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 6541 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 6601 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 6661 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 6721 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 6781 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 6841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 6901 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 6961 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 7021 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 7081 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 7141 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 7201 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 7261 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 7321 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 7381 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 7441 cacatgt //