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
//

