LOCUS Exported 12372 bp ds-DNA circular SYN 29-OCT-2015 DEFINITION Ligation of pLKO5d.EFS.NLS-SpCas9-NLS.P2A-PAC.pre* into pL40C-CRISPR.EFS.mNeon NL-SL*. ACCESSION . VERSION . KEYWORDS pL40C-CRISPR.EFS.PAC NL SL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12372) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 29, 2015 from SnapGene Viewer 2.6.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12372 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 430..811 /note="CMV IE enhancer" misc_feature 826..1058 /note="SV40 enhancer" misc_feature 1075..1487 /note="RSV/5'LTR" /note="LTR" misc_feature 1538..1675 /note="psi" misc_feature 1642..1996 /note="gag'" misc_feature 2911..3028 /note="cPPT" misc_feature 3079..3327 /note="U6 Prom" misc_feature 3355..3446 /note="sgRNA" misc_feature 3359 /note="mod T-A" misc_feature 3367..3371 /note="elong stem" misc_feature 3376..3380 /note="elong stem comp" misc_feature 3390 /note="mod A-T" misc_feature 3469..3713 /note="EFS" misc_feature 3729..7953 /note="SpCas9'" misc_feature 7959..8024 /note="P2A" misc_feature 8025..8624 /note="PuroR" misc_feature 8640..9228 /note="WPRE" /note="misc_RNA" misc_feature 9299..9534 /note="3'\LTR\SIN" /note="LTR" misc_feature 9611..9741 /note="SV40\polyA" /note="polyA_signal" misc_feature 10088..10518 /note="f1\Ori" /note="rep_origin" misc_feature 10889..11695 /note="AmpR" misc_feature complement(11828..64) /note="pUC\Ori" /note="rep_origin" ORIGIN 1 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 61 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg 181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt 241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt 301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc 361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cggtaccgtc 421 gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat 481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg 541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa 601 cgccaatagg gactttccat tgacgtcaat gggtggacta tttacggtaa actgcccact 661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta 721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt 781 acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct gaaataacct 841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt 901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 1021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg 1081 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc 1141 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt 1201 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg 1261 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta 1321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa 1381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac 1441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca 1501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg 1561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta 1621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta 1681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac 1741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa 1801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag 1861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag 1921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga 1981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat 2041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc 2101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg 2161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg 2221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct 2281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca 2341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc 2401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct 2461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac 2521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa 2581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg 2641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt 2701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag 2761 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga 2821 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt 2881 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta 2941 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca 3001 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt 3061 ttggttaatt aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga 3121 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta 3181 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt 3241 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta 3301 tatatcttgt ggaaaggacg aaacaccgga gacgtatgtc gacaactcgt ctctgtttaa 3361 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa 3421 aagtggcacc gagtcggtgc ttttttgaat tctcctctag aactcgagtg gctccggtgc 3481 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg 3541 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta 3601 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt 3661 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg 3721 ccaccatgga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg 3781 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca 3841 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg 3901 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg 3961 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca 4021 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg 4081 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg 4141 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca 4201 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga 4261 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc 4321 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc 4381 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg 4441 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga 4501 agaatggcct gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga 4561 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg 4621 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca 4681 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca 4741 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc 4801 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc 4861 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca 4921 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga 4981 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga 5041 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga 5101 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc 5161 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca 5221 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc 5281 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc 5341 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa 5401 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca 5461 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt 5521 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat 5581 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg 5641 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg 5701 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc 5761 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc 5821 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca 5881 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt 5941 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta 6001 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc 6061 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac 6121 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc 6181 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt 6241 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt 6301 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca 6361 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag 6421 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc 6481 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg 6541 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga 6601 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga 6661 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca 6721 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg 6781 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca 6841 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg 6901 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca 6961 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt 7021 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata 7081 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga 7141 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct 7201 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt 7261 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct 7321 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc 7381 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc 7441 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga 7501 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc 7561 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca 7621 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct 7681 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta 7741 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga 7801 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg 7861 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaagcgt cctgctgcta 7921 ctaagaaagc tggtcaagct aagaaaaaga aagctagcgg cagcggcgcc accaacttca 7981 gcctgctgaa gcaggccggc gacgtggagg agaaccccgg ccccatgacc gagtacaagc 8041 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg 8101 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg 8161 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg 8221 tcgcggacga cggcgcggcc gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg 8281 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc 8341 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg 8401 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc 8461 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc 8521 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg 8581 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaacgcgt taagtcgaca 8641 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc 8701 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta 8761 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt 8821 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg 8881 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta 8941 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt 9001 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg 9061 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca 9121 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc 9181 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag 9241 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact 9301 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct 9361 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa 9421 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc 9481 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta 9541 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga 9601 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 9661 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 9721 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc 9781 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat 9841 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt 9901 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg 9961 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg 10021 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt 10081 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg 10141 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 10201 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 10261 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 10321 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 10381 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 10441 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 10501 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc 10561 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 10621 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 10681 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 10741 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 10801 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 10861 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 10921 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 10981 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 11041 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 11101 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 11161 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 11221 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 11281 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 11341 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 11401 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 11461 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 11521 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 11581 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 11641 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 11701 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 11761 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 11821 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 11881 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 11941 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 12001 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 12061 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 12121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 12181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 12241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 12301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 12361 ctggcctttt gc //