LOCUS       pSAMca116_-_pHKO30-sgRN 8387 bp ds-DNA     linear       10-FEB-2016
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    complement(790..807)
                     /label="XRP008 partial binding site"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     LTR             6672..7084
                     /label="RSV/5'LTR"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    287..291
                     /label="MS2 stem"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    643..673
                     /label="TGCAAAGATGGATAAAGTTTTAAACAGAGAG"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_RNA        2528..3116
                     /label="WPRE"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     variation       1031..1031
                     /label="C in all sequences"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    352..385
                     /label="MS2 binding loop"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    282..315
                     /label="MS2 binding loop"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     rep_origin      3976..4406
                     /label="f1 Ori"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    301..305
                     /label="MS2 stem"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(2229..2247)
                     /label="AGGCCTTCCATCTGTTGCT"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    1742..1765
                     /label="EF1A-F seq primer"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    7749..7990
                     /label="RRE"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    7239..7603
                     /label="gag"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    406..411
                     /label="U6 terminator"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     2885..2904
                     /label="HKO034 (Rv)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    1913..1934
                     /label="ATGACCGAGTACAAGCCCACGG"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    357..361
                     /label="MS2 stem"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     polyA_signal    3499..3629
                     /label="SV40 polyA"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     rep_origin      complement(5709..6324)
                     /label="pUC Ori"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    282..285
                     /label="GC tract"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     promoter        1..245
                     /label="Human U6 Promoter"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    352..355
                     /label="GC tract"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     LTR             3187..3422
                     /label="3' LTR SIN"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     2409..2428
                     /label="HKO033"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    365..368
                     /label="loop"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    2299..2299
                     /label="Destroy BsmBI site in Puro"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     promoter        643..1900
                     /label="EF-1a"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    1913..2512
                     /label="Puromycin"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    7135..7272
                     /label="psi"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(2496..2512)
                     /label="TCAGGCACCGGGCTTGC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    295..298
                     /label="loop"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    371..375
                     /label="MS2 stem"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4777..5583
                     /label="AmpR"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     complement(2229..2247)
                     /label="HKO032"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    382..385
                     /label="GC tract"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    complement(391..411)
                     /label="AAAAAAAGCACCGACTCGGTGC"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    467..584
                     /label="cPPT"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    complement(783..800)
                     /label="TCAATTGCCGACCCCTCC"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    270..405
                     /label="chimeric guide RNA scaffold"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     842..861
                     /label="HKO038 (Rv)"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    312..315
                     /label="GC tract"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt GTGGAAAGGA
      241 CGAAACACCG GAGACGGGAT ACCGTCTCTg ttttagagct aggccAACAT GAGGATCACC
      301 CATGTCTGCA Gggcctagca agttaaaata aggctagtcc gttatcaact tggccAACAT
      361 GAGGATCACC CATGTCTGCA Gggccaagtg gcaccgagtc ggtgcTTTTT TGGATCCAAG
      421 CTTGGCGTAA CTAGATCTTG AGACAAATGG CAGTATTCAT CCACAATTTT AAAAGAAAAG
      481 GGGGGATTGG GGGGTACAGT GCAGGGGAAA GAATAGTAGA CATAATAGCA ACAGACATAC
      541 AAACTAAAGA ATTACAAAAA CAAATTACAA AAATTCAAAA TTTTCGGGTT TATTACAGGG
      601 ACAGCAGAGA TCCACTTTGG CGCCGGCTCG AGGGGGcccg ggtgcaaaga tggataaagt
      661 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      721 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      781 ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa
      841 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      901 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      961 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     1021 ttacttccac tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     1081 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     1141 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     1201 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     1261 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     1321 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     1381 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     1441 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     1501 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     1561 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     1621 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     1681 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1741 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1801 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1861 agtggttcaa agtttttttc ttccatttca ggtgtcgtga cgtacggcca ccatgaccga
     1921 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     1981 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     2041 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     2101 ggtgtgggtc gcggacgacg gcgccgccgt ggcggtctgg accacgccgg agagcgtcga
     2161 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     2221 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     2281 gttcctggcc accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     2341 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     2401 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     2461 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta
     2521 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2581 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2641 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2701 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2761 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2821 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2881 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2941 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3001 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3061 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     3121 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     3181 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     3241 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     3301 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     3361 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     3421 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     3481 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3601 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     3661 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3721 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3781 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
     3841 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3901 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3961 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     4021 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4081 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4141 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4201 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4261 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4321 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4381 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4441 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4501 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4561 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4621 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4681 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4741 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4801 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4861 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4921 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4981 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5041 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5101 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5161 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5221 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5281 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5341 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5401 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5461 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5521 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5581 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5641 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5701 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5761 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5821 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5881 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5941 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6001 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6061 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6121 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6181 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6241 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6301 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6361 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6481 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6541 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6601 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     6661 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
     6721 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
     6781 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
     6841 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
     6901 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6961 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     7021 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     7081 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     7141 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     7201 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     7261 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     7321 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     7381 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     7441 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     7501 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     7561 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     7621 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     7681 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     7741 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     7801 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     7861 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7921 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7981 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     8041 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     8101 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     8161 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     8221 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     8281 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     8341 caccattatc gtttcagacc cacctcccaa ccccgagggg acccaga
//