LOCUS       pZac-SMVP-NLS           7479 bp    DNA     circular     03-AUG-2016
Addgene 80937
Wei Leong Chew
George Church lab
FEATURES             Location/Qualifiers
     repeat_region   2012..2141
                     /dnas_title="ITR"
                     /vntifkey="34"
                     /label=ITR
     repeat_region   complement(6550..6679)
                     /dnas_title="ITR"
                     /vntifkey="34"
                     /label=ITR
     promoter        2470..3022
                     /label=CMV_immearly_promoter
     misc_feature    2525..2812
                     /label=CAG_enhancer
     misc_feature    2308..3049
                     /note="CMV IE"
     protein_bind    2314..2327
                     /note="NF1"
     protein_bind    2361..2374
                     /note="NF1"
     protein_bind    2415..2428
                     /note="NF1"
     promoter        2213..2284
                     /label=SV40 enhancer
     protein_bind    2218..2228
                     /note="NFkB2"
     promoter        2968..3049
                     /label=CMV_promoter
     misc_feature    2979..2999
                     /label=CMV_fwd_primer
     TATA_signal     3014..3019
                     /note=""
     misc_feature    3120..3252
                     /label=chim_int
     gene            3288..5474
                     /note="hCas9"
     misc_feature    3294..3317
                     /note="SV40 NLS"
     misc_feature    5472..5474
                     /note=""
     misc_feature    5475..5780
                     /note="N-intein"
     misc_feature    complement(5794..5813)
                     /label=EBV_rev_primer
     3'UTR           5794..5995
                     /note="SV40 pA"
     polyA_signal    5893..5898
                     /note=""
     promoter        6066..6384
                     /note="U6 promoter"
     misc_feature    6413..6495
                     /note="Sp gRNA scaffold"
     misc_feature    6385..6412
                     /note="Spacer"
     source          1..7479
                     /dnas_title="pZac-SMVP-NLS SphCas9- Rma Int-N trun -
                     SV40pA - U6 -Sp gRNA (mMstn4)(Addgene 80937)"
ORIGIN      
        1 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
       61 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aatagtggta
      121 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
      181 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
      241 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
      301 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
      361 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
      421 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
      481 acgacgagcg tgacaccacg atgcctgtag taatggtaac aacgttgcgc aaactattaa
      541 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
      601 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
      661 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
      721 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
      781 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
      841 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
      901 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
      961 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1021 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1081 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1141 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1201 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1261 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1321 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1381 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1441 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1501 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1561 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1621 tttgctgcgg ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1681 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     1741 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     1801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     1861 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     1921 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     1981 atgaccatga ttacgccaga tttaattaag gctgcgcgct cgctcgctca ctgaggccgc
     2041 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     2101 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc
     2161 gccatgctac ttatctacCT GCgGccGcCA TTATGGAATG TGTGTCAGTT AGGGTGTGGA
     2221 AAGTCCCCAG GCTCCCCAGC AGGCAGAAGT ATGCAAAGCA TGCATCTCAA TTAGTCAGCA
     2281 ACCAgtcccg gtctcctccc atgcatgtca atattggcca ttagccatat tattcattgg
     2341 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     2401 tatgtacatt tatattggct catgtccaat atgaccgcca tgttggcatt gattattgac
     2461 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     2521 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2581 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2641 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     2701 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     2761 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     2821 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     2881 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     2941 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     3001 acggtgggag gtctatataa gcagaggtcg tttagtgaac cgtcagatca ctagtagctt
     3061 tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgctcgac tgatcacagg
     3121 taagtatcaa ggttacaaga caggtttaag gaggccaata gaaactgggc ttgtcgagac
     3181 agagaagatt cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     3241 ttctctccac aggGGTACCG AAGCCGCTAG CGCTACCGGT CGCCACCATG GCCCCAAAGA
     3301 AGAAGCGGAA GGTCGGTATC CACGGAGTCC CAGCAGCCGA CAAGAAGTAC TCCATTGGGC
     3361 TCGATATCGG CACAAACAGC GTCGGCTGGG CCGTCATTAC GGACGAGTAC AAGGTGCCGA
     3421 GCAAAAAATT CAAAGTTCTG GGCAATACCG ATCGCCACAG CATAAAGAAG AACCTCATTG
     3481 GCGCCCTCCT GTTCGACTCC GGGGAaACGG CCGAAGCCAC GCGGCTCAAA AGAACAGCAC
     3541 GGCGCAGATA TACCCGCAGA AAGAATCGGA TCTGCTACCT GCAGGAGATC TTTAGTAATG
     3601 AGATGGCTAA GGTGGATGAC TCTTTCTTCC ATAGGCTGGA GGAGTCCTTT TTGGTGGAGG
     3661 AGGATAAAAA GCACGAGCGC CACCCAATCT TTGGCAATAT CGTGGACGAG GTGGCGTACC
     3721 ATGAAAAGTA CCCAACCATA TATCATCTGA GGAAGAAGCT TGTAGACAGT ACTGATAAGG
     3781 CTGACTTGCG GTTGATCTAT CTCGCGCTGG CGCATATGAT CAAATTTCGG GGACACTTCC
     3841 TCATCGAGGG GGACCTGAAC CCAGACAACA GCGATGTCGA CAAACTCTTT ATCCAACTGG
     3901 TTCAGACTTA CAATCAGCTT TTCGAAGAGA ACCCGATCAA CGCATCCGGA GTTGACGCCA
     3961 AAGCAATCCT GAGCGCTAGG CTGTCCAAAT CCCGGCGGCT CGAAAACCTC ATCGCACAGC
     4021 TCCCTGGGGA GAAGAAGAAC GGCCTGTTTG GTAATCTTAT CGCCCTGTCA CTCGGGCTGA
     4081 CCCCCAACTT TAAATCTAAC TTCGACCTGG CCGAAGATGC CAAGCTTCAA CTGAGCAAAG
     4141 ACACCTACGA TGATGATCTC GACAATCTGC TGGCCCAGAT CGGCGACCAG TACGCAGACC
     4201 TTTTTTTGGC GGCAAAGAAC CTGTCAGACG CCATTCTGCT GAGTGATATT CTGCGAGTGA
     4261 ACACGGAGAT CACCAAAGCT CCGCTGAGCG CTAGTATGAT CAAGCGCTAT GATGAGCACC
     4321 ACCAAGACTT GACTTTGCTG AAGGCCCTTG TCAGACAGCA ACTGCCTGAG AAGTACAAGG
     4381 AAATTTTCTT CGATCAGTCT AAAAATGGCT ACGCCGGATA CATTGACGGC GGAGCAAGCC
     4441 AGGAGGAATT TTACAAATTT ATTAAGCCCA TCTTGGAAAA AATGGACGGC ACCGAGGAGC
     4501 TGCTGGTAAA GCTTAACAGA GAAGATCTGT TGCGCAAACA GCGCACTTTC GACAATGGAA
     4561 GCATCCCCCA CCAGATTCAC CTGGGCGAAC TGCACGCTAT CCTCAGGCGG CAAGAGGATT
     4621 TCTACCCCTT TTTGAAAGAT AACAGGGAAA AGATTGAGAA AATCCTCACA TTTCGGATAC
     4681 CCTACTATGT AGGCCCCCTC GCCCGGGGAA ATTCCAGATT CGCGTGGATG ACTCGCAAAT
     4741 CAGAAGAgAC CATCACTCCC TGGAACTTCG AGGAAGTCGT GGATAAGGGG GCCTCTGCCC
     4801 AGTCCTTCAT CGAAAGGATG ACTAACTTTG ATAAAAATCT GCCTAACGAA AAGGTGCTTC
     4861 CTAAACACTC TCTGCTGTAC GAGTACTTCA CAGTTTATAA CGAGCTCACC AAGGTCAAAT
     4921 ACGTCACAGA AGGGATGAGA AAGCCAGCAT TCCTGTCTGG AGAGCAGAAG AAAGCTATCG
     4981 TGGACCTCCT CTTCAAGACG AACCGGAAAG TTACCGTGAA ACAGCTCAAA GAAGACTATT
     5041 TCAAAAAGAT TGAATGTTTC GACTCTGTTG AAATCAGCGG AGTGGAGGAT CGCTTCAACG
     5101 CATCCCTGGG AACGTATCAC GATCTCCTGA AAATCATTAA AGACAAGGAC TTCCTGGACA
     5161 ATGAGGAGAA CGAGGACATT CTTGAGGACA TTGTCCTCAC CCTTACGTTG TTTGAAGATA
     5221 GGGAGATGAT TGAAGAACGC TTGAAAACTT ACGCTCATCT CTTCGACGAC AAAGTCATGA
     5281 AACAGCTCAA GAGGCGCCGA TATACAGGAT GGGGGCGGCT GTCAAGAAAA CTGATCAATG
     5341 GGATCCGAGA CAAGCAGAGT GGAAAGACAA TCCTGGATTT TCTTAAGTCC GATGGATTTG
     5401 CCAACCGGAA CTTCATGCAG TTGATCCATG ATGACTCTCT CACCTTTAAG GAGGACATCC
     5461 AGAAAGCACA AGTTtgtctg gctggcgata ctctcattac cctggccgat ggacgacgag
     5521 tgcctattag agaactggtg tcacagcaga atttttccgt gtgggctctg aatcctcaga
     5581 cttaccgcct ggagagggct agagtgagta gagctttctg taccggcatc aaacctgtgt
     5641 accgcctcac cactagactg gggagatcca ttagggccac tgccaaccac cgatttctca
     5701 cacctcaggg ctggaaacga gtcgatgaac tccagcctgg agattacctg gctctgccta
     5761 ggagaatccc tactgcctcc tgaTCTCGAG CTcgatgagt ttggacaaac cacaactaga
     5821 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5881 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     5941 cagggggagg tgtgggaggt tttttaaaGC AAGTAAAACC TCTACAAATG TGGTActtaa
     6001 gagggggaga ccaaagggcg agacgttaag gcctcacgtg acatgtgagc aaaaggccag
     6061 caaaaTGTAC AAAAAAGCAG GCTTTAAAGG AACCAATTCA GTCGACTGGA TCCGGTACCA
     6121 AGGTCGGGCA GGAAGAGGGC CTATTTCCCA TGATTCCTTC ATATTTGCAT ATACGATACA
     6181 AGGCTGTTAG AGAGATAATT AGAATTAATT TGACTGTAAA CACAAAGATA TTAGTACAAA
     6241 ATACGTGACG TAGAAAGTAA TAATTTCTTG GGTAGTTTGC AGTTTTAAAA TTATGTTTTA
     6301 AAATGGACTA TCATATGCTT ACCGTAACTT GAAAGTATTT CGATTTCTTG GCTTTATATA
     6361 TCTTGTGGAA AGGACGAAAC ACCGgaatcc cggtgctgcc gctaccccct caGTTTTAGA
     6421 GCTAGAAATA GCAAGTTAAA ATAAGGCTAG TCCGTTATCA ACTTGAAAAA GTGGCACCGA
     6481 GTCGGTGCTT TTTTTtCTAG AggGCGGCCG CAgtagataa gtagcatggc gggttaatca
     6541 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     6601 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     6661 tgagcgagcg agcgcgcagc cttaattaac ctaattcact ggccgtcgtt ttacaacgtc
     6721 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     6781 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     6841 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6901 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6961 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7021 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7081 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7141 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7201 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7261 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttataatt tcaggtggca
     7321 tctttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     7381 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     7441 gtatgagtat tcaacatttc cgtgtcgccc ttattccct
//

