LOCUS       140708_SpSelfcut        7624 bp    DNA     linear       25-JUL-2017
FEATURES             Location/Qualifiers
     misc_feature    7331..7407
                     /label=SpCas9 gRNA scaffold
     terminator      7408..7413
                     /note="U6 terminator"
     variation       7358..7358
                     /mod_base=A > C
     variation       7359..7359
                     /mod_base=A > C
     gene            590..1624
                     /note="Hygromycin"
     PCR_primer      complement(1602..1621)
                     /label=PB-Cas9-Hyg_n8647_R
                     /note="Ordered sequence:       PB-Cas9-Hyg_n8647_R:  
                     CTAAGCGTAATCTGGAACATCGTATGGGTA TTCCTTTGCCCTCGGACGAG"
     gene            3141..4001
                     /note="Amp"
     promoter        7061..7302
                     /note="U6 Promoter"
     PCR_primer      complement(7364..7385)
                     /note="SP-Hyg-LV_R1"
     PCR_primer      7229..7251
                     /note="Sp-Hyg-LV_F1"
     primer_bind     7232..7251
                     /label=pLKO.1 5'
                     /note="Human U6 promoter, forward primer"
     variation       7332..7332
                     /mod_base=T > G
     variation       7333..7333
                     /mod_base=T > G
     misc_feature    7310..7310
                     /note="TSS"
     misc_feature    7311..7330
                     /label=V6 Protospacer
                     /note=""
     source          1..7624
                     /dnas_title="140708_SpSelfcuttingGuide_d1"
ORIGIN      
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccaccgg atcaagcttg ccagctgggg
      541 cgccctctgg taaggttggg aagccctgca gatctgatcc cggggggcaA TGGATAGATC
      601 CGGAAAGCCT GAACTCACCG CGACGTCTGT CGAGAAGTTT CTGATCGAAA AGTTCGACAG
      661 CGTCTCCGAC CTGATGCAGC TCTCGGAGGG CGAAGAATCT CGTGCTTTCA GCTTCGATGT
      721 AGGAGGGCGT GGATATGTCC TGCGGGTAAA TAGCTGCGCC GATGGTTTCT ACAAAGATCG
      781 TTATGTTTAT CGGCACTTTG CATCGGCCGC GCTCCCGATT CCGGAAGTGC TTGACATTGG
      841 GGAATTTAGC GAGAGCCTGA CCTATTGCAT CTCCCGCCGT GCACAGGGTG TCACGTTGCA
      901 AGACCTGCCT GAAACCGAAC TGCCCGCTGT TCTGCAGCCG GTCGCGGAGG CCATGGATGC
      961 GATCGCTGCG GCCGATCTTA GCCAGACGAG CGGGTTCGGC CCATTCGGAC CGCAAGGAAT
     1021 CGGTCAATAC ACTACATGGC GTGATTTCAT ATGCGCGATT GCTGATCCCC ATGTGTATCA
     1081 CTGGCAAACT GTGATGGACG ACACCGTCAG TGCGTCCGTC GCGCAGGCTC TCGATGAGCT
     1141 GATGCTTTGG GCCGAGGACT GCCCCGAAGT CCGGCACCTC GTGCACGCGG ATTTCGGCTC
     1201 CAACAATGTC CTGACGGACA ATGGCCGCAT AACAGCGGTC ATTGACTGGA GCGAGGCGAT
     1261 GTTCGGGGAT TCCCAATACG AGGTCGCCAA CATCTTCTTC TGGAGGCCGT GGTTGGCTTG
     1321 TATGGAGCAG CAGACGCGCT ACTTCGAGCG GAGGCATCCG GAGCTTGCAG GATCGCCGCG
     1381 GCTCCGGGCG TATATGCTCC GCATTGGTCT TGACCAACTC TATCAGAGCT TGGTTGACGG
     1441 CAATTTCGAT GATGCAGCTT GGGCGCAGGG TCGATGCGAC GCAATCGTCC GATCCGGAGC
     1501 CGGGACTGTC GGGCGTACAC AAATCGCCCG CAGAAGCGCG GCCGTCTGGA CCGATGGCTG
     1561 TGTAGAAGTA CTCGCCGATA GTGGAAACCG ACGCCCCAGC ACTCGTCCGA GGGCAAAGGA
     1621 ATAGAGTAGA TGCCGACCGG GATCTATCGA TACCGTCGAC CTCGAGGGGG GGCCcggtac
     1681 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     1741 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     1801 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     1861 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     1921 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     1981 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     2041 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2101 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2161 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     2221 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2281 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2341 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     2401 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2461 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2521 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2581 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2641 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2701 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2761 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2821 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2881 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2941 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3001 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3061 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3121 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3181 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3241 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3301 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3361 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3421 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3481 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3541 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3601 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3661 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3721 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3781 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3841 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3901 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3961 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4021 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4081 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4141 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4201 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4261 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4321 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4381 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4441 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4501 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4561 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4621 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4681 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4741 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4801 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4861 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4921 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4981 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5041 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     5101 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     5161 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
     5221 aagcttaatg tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta
     5281 gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg
     5341 gtacgatcgt gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac
     5401 tgaattgccg cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc
     5461 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     5521 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     5581 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
     5641 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
     5701 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
     5761 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
     5821 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
     5881 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
     5941 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
     6001 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
     6061 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
     6121 aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat
     6181 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     6241 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     6301 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     6361 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     6421 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     6481 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     6541 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     6601 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     6661 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     6721 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     6781 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     6841 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     6901 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     6961 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta
     7021 tcgatcacga gactagcctc gagcggccgc ccccttcacc gagggcctat ttcccatgat
     7081 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
     7141 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
     7201 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
     7261 gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccG GTTCCAGAGA
     7321 TACTGACGTG GGGTTAGAGC TAGAAATAGC AAGTTAACCT AAGGCTAGTC CGTTATCAAC
     7381 TTGAAAAAGT GGCACCGAGT CGGTGCTTTT TTTGAATTCt cgacctcgag acaaatggca
     7441 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     7501 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     7561 attcaaaatt ttcgggttta ttacagggac agcagagatc cactttggcc gcggctcgag
     7621 gggg
//

