LOCUS pSimple\II-NLS-N 8501 bp DNA circular 7-AUG-2013 DEFINITION general stem cell expression vector. SOURCE ORGANISM COMMENT This file is created by Vector NTI http://www.invitrogen.com/ COMMENT VNTDATE|669758538| COMMENT VNTDBDATE|669759453| COMMENT LSOWNER| COMMENT VNTNAME|pSimple II-NLS-NmCas9-HA-NLS(s)| COMMENT VNTAUTHORNAME|Demo User| COMMENT VNTREPLTYPE|Plasmid COMMENT VNTEXTCHREPL|Bacteria COMMENT VNTEXTCHREPL|Animal/Other Eukaryotic COMMENT VNTUDF|Created by|4|Takashi Kameda| COMMENT Vector_NTI_Display_Data_(Do_Not_Edit!) 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COMMENT (CWideLine COMMENT (CWidget 0 (3 #3# 0) 1 2 0 0 #58# 0 100) COMMENT (LOGPEN 0 0 3355545) 1 (LOGBRUSH 0 6724095 0) COMMENT 1 0.841246 1.33935 1 0.082322) 0.8 1.8 0) COMMENT #122=(CLabel (CWidget 0 (0 0) 1 2 0 0 #55# 14 100) COMMENT (LOGPEN 0 0 0) 1 COMMENT (LOGFONT 69 26 0 0 700 0 0 0 0 3 2 1 34 "Arial") COMMENT 1.74233 0.666667 0 "pUC" "@N" 1 -12.0458 COMMENT 5.22699 0 -10 2.03141 1.06566 #121#)) COMMENT (CObjectList))) (CObjectList)) COMMENT #123=(CGroupWidget (CWidget 14 (16 0) 1 2 0 0 Nil -1181 100) COMMENT (CObjectList) (CObjectList)) COMMENT #124=(CGroupWidget (CWidget 11 (0 0) 1 2 0 0 Nil -134 100) COMMENT (CObjectList) (CObjectList)) COMMENT #125=(CGroupWidget (CWidget 12 (0 0) 1 2 0 0 Nil -810 100) COMMENT (CObjectList) (CObjectList))) (CObjectList))) COMMENT (CSeqView 10 10 (CObjectList) (CObList) 1 (CObList)) (CObList) 64385704 COMMENT (CStringList) 64369144 64369864 (CObList))) FEATURES Location/Qualifiers promoter 231..1400 /dnas_title="EF1a P" /vntifkey="29" /label=EF1a P /note="EF1 alpha promoter" polyA_signal 4693..4899 /dnas_title="BGH polyA" /vntifkey="25" /label=BGH polyA /note="BGH poly A signal (Bovine growth hormone)" rep_origin 5124..6246 /dnas_title="mini OriP" /vntifkey="33" /label=mini OriP /note="mini OriP (short version of OriP, allows plasmid replication in stem cells)" rep_origin 6627..7301 /dnas_title="pUC" /vntifkey="33" /label=pUC /note="pUC bacterial origin of replication" CDS complement(7448..8305) /dnas_title="AP(R)" /vntifkey="4" /label=AP(R) /note="ampicillin resistance" CDS 1424..4666 /dnas_title="NmCas9" /vntifkey="4" /label=NmCas9 misc_feature 4667..4690 /note="FLAG tag" source 1..8441 /dnas_title="pSimple II-NmCas9-FLAG" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcggg ggtgaggctc cggtgcccgt cgtgaggctc 241 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg 301 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt 361 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt 421 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg 481 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc 541 cacctggctc cagtacgtga ttcttgatcc cgagctggag ccaggggcgg gccttgcgct 601 ttaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc 661 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt 721 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg 781 ggccaggatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt 841 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga 901 cgggggtcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1021 cggaaagatg gccgcttccc ggccctgctc cagggggctc aaaatggagg acgcggcgct 1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaagg ggcctttccg tcctcagccg 1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctgga 1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1381 gtttttttct tccatttcag gtgtcgtgaa caccgccacc atggctgcct tcaaacctaa 1441 ttcaatcaac tacatcctcg gcctcgatat cggcatcgca tccgtcggct gggcgatggt 1501 agaaattgac gaagaagaaa accccatccg cctgattgat ttgggcgtgc gcgtatttga 1561 gcgtgccgaa gtaccgaaaa caggcgactc ccttgccatg gcaaggcgtt tggcgcgcag 1621 tgttcgccgc ctgacccgcc gtcgcgccca ccgcctgctt cggacccgcc gcctattgaa 1681 acgcgaaggc gtattacaag ccgccaattt tgacgaaaac ggcttgatta aatccttacc 1741 gaatacacca tggcaacttc gcgcagccgc attagaccgc aaactgacgc ctttagagtg 1801 gtcggcagtc ttgttgcatt taatcaaaca tcgcggctat ttatcgcaac ggaaaaacga 1861 gggcgaaact gccgataagg agcttggcgc tttgcttaaa ggcgtagccg gcaatgccca 1921 tgccttacag acaggcgatt tccgcacacc ggccgaattg gctttaaata aatttgagaa 1981 agaaagcggc catatccgca atcagcgcag cgattattcg catacgttca gccgcaaaga 2041 tttacaggcg gagctgattt tgctgtttga aaaacaaaaa gaatttggca atccgcatgt 2101 ttcaggcggc cttaaagaag gtattgaaac cctactgatg acgcaacgcc ctgccctgtc 2161 cggcgatgcc gttcaaaaaa tgttggggca ttgcaccttc gaaccggcag agccgaaagc 2221 cgctaaaaac acctacacag ccgaacgttt catctggctg accaagctga acaacctgcg 2281 tattttagag caaggcagcg agcggccatt gaccgatacc gaacgcgcca cgcttatgga 2341 cgagccatac agaaaatcca aactgactta cgcacaagcc cgtaagctgc tgggtttaga 2401 agataccgcc tttttcaaag gcttgcgcta tggtaaagac aatgccgaag cctcaacatt 2461 gatggaaatg aaggcctacc atgccatcag ccgtgcactg gaaaaagaag gattgaaaga 2521 caaaaaatcc ccattaaacc tttctcccga attacaagac gaaatcggca cggcattctc 2581 cctgttcaaa accgatgaag acattacagg ccgtctgaaa gaccgtatac agcccgaaat 2641 cttagaagcg ctgttgaaac acatcagctt cgataagttc gtccaaattt ccttgaaagc 2701 attgcgccga attgtgcctc taatggaaca aggcaaacgt tacgatgaag cctgcgccga 2761 aatctacgga gaccattacg gcaagaagaa tacggaagaa aagatttatc tgccgccgat 2821 tcccgccgac gaaatccgca accccgtcgt cttgcgcgcc ttatctcaag cacgtaaggt 2881 cattaacggc gtggtacgcc gttacggctc cccagctcgt atccatattg aaactgcaag 2941 ggaagtaggt aaatcgttta aagaccgcaa agaaattgag aaacgccaag aagaaaaccg 3001 caaagaccgg gaaaaagccg ccgccaaatt ccgagagtat ttccccaatt ttgtcggaga 3061 acccaaatcc aaagatattc tgaaactgcg cctgtacgag caacaacacg gcaaatgcct 3121 gtattcgggc aaagaaatca acttaggccg tctgaacgaa aaaggctatg tcgaaatcga 3181 ccatgccctg ccgttctcgc gcacatggga cgacagtttc aacaataaag tactggtatt 3241 gggcagcgaa aaccaaaaca aaggcaatca aaccccttac gaatacttca acggcaaaga 3301 caacagccgc gaatggcagg aatttaaagc gcgtgtcgaa accagccgtt tcccgcgcag 3361 taaaaaacaa cggattctgc tgcaaaaatt cgatgaagac ggctttaaag aacgcaatct 3421 gaacgacacg cgctacgtca accgtttcct gtgtcaattt gttgccgacc gtatgcggct 3481 gacaggtaaa ggcaagaaac gtgtctttgc atccaacgga caaattacca atctgttgcg 3541 cggcttttgg ggattgcgca aagtgcgtgc ggaaaacgac cgccatcacg ccttggacgc 3601 cgtcgtcgtt gcctgctcga ccgttgccat gcagcagaaa attacccgtt ttgtacgcta 3661 taaagagatg aacgcgtttg acggtaaaac catagacaaa gaaacaggag aagtgctgca 3721 tcaaaaaaca cacttcccac aaccttggga atttttcgca caagaagtca tgattcgcgt 3781 cttcggcaaa ccggacggca aacccgaatt cgaagaagcc gataccctag aaaaactgcg 3841 cacgttgctt gccgaaaaat tatcatctcg ccccgaagcc gtacacgaat acgttacgcc 3901 actgtttgtt tcacgcgcgc ccaatcggaa gatgagcggg caagggcata tggagaccgt 3961 caaatccgcc aaacgactgg acgaaggcgt cagcgtgttg cgcgtaccgc tgacacagtt 4021 aaaactgaaa gacttggaaa aaatggtcaa tcgggagcgc gaacctaagc tatacgaagc 4081 actgaaagca cggctggaag cacataaaga cgatcctgcc aaagcctttg ccgagccgtt 4141 ttacaaatac gataaagcag gcaaccgcac ccaacaggta aaagccgtac gcgtagagca 4201 agtacagaaa accggcgtat gggtgcgcaa ccataacggt attgccgaca acgcaaccat 4261 ggtgcgcgta gatgtgtttg agaaaggcga caagtattat ctggtaccga tttacagttg 4321 gcaggtagcg aaagggattt tgccggatag ggctgttgta caaggaaaag atgaagaaga 4381 ttggcaactt attgatgata gtttcaactt taaattctca ttacacccta atgatttagt 4441 cgaggttata acaaaaaaag ctagaatgtt tggttacttt gccagctgcc atcgaggcac 4501 aggtaatatc aatatacgca ttcatgatct tgatcataaa attggcaaaa atggaatact 4561 ggaaggtatc ggcgtcaaaa ccgccctttc attccaaaaa taccaaattg acgaactggg 4621 caaagaaatc agaccatgcc gtctgaaaaa acgcccgcct gtccgtgatt ataaggatga 4681 tgatgataag taactcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt 4741 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc 4801 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt 4861 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat 4921 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgac cgatgccctt 4981 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc 5041 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgcctggcc ggggtccccc 5101 ggaaactcgg ccgtggtgac catgcaggaa aaggacaagc agcgaaaatt cacgccccct 5161 tgggaggtgg cggcatatgc aaaggatagc actcccactc tactactggg tatcatatgc 5221 tgactgtata tgcatgagga tagcatatgc tacccggata cagattagga tagcatatac 5281 tacccagata tagattagga tagcatatgc tacccagata tagattagga tagcctatgc 5341 tacccagata taaattagga tagcatatac tacccagata tagattagga tagcatatgc 5401 tacccagata tagattagga tagcctatgc tacccagata tagattagga tagcatatgc 5461 tacccagata tagattagga tagcatatgc tatccagata tttgggtagt atatgctacc 5521 cagatataaa ttaggatagc atatactacc ctaatctcta ttaggatagc atatgctacc 5581 cggatacaga ttaggatagc atatactacc cagatataga ttaggatagc atatgctacc 5641 cagatataga ttaggatagc ctatgctacc cagatataaa ttaggatagc atatactacc 5701 cagatataga ttaggatagc atatgctacc cagatataga ttaggatagc ctatgctacc 5761 cagatataga ttaggatagc atatgctatc cagatatttg ggtagtatat gctacccatg 5821 gcaacattag cccaccgtgc tctcagcgac ctcgtgaata tgaggaccaa caaccctgtg 5881 cttggcgctc aggcgcaagt gtgtgtaatt tgtcctccag atcgcagcaa tcgcgcccct 5941 atcttggccc gcccacctac ttatgcaggt attccccggg gtgccattag tggttttgtg 6001 ggcaagtggt ttgaccgcag tggttagcgg ggttacaatc agccaagtta ttacaccctt 6061 attttacagt ccaaaaccgc agggcggcgt gtgggggctg acgcgatcgc tgttccttag 6121 gaccctttta ctaaccctaa ttcgatagca tatgcttccc gttgggtaac atatgctatt 6181 gaattagggt tagtctggat agtatatact actacccggg aagcatatgc tacccgttta 6241 gggttatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg 6301 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 6361 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 6421 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 6481 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 6541 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 6601 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 6661 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 6721 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 6781 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 6841 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc 6901 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 6961 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 7021 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7081 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 7141 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7201 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7261 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 7321 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 7381 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 7441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 7501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 7561 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 7621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 7681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 7741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 7801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 7861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 7921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 7981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 8101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 8161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 8221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 8281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 8341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 8401 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c //