LOCUS PXL 8548 bp ds-DNA circular 20-APR-2016 DEFINITION . FEATURES Location/Qualifiers CDS 3428..4288 /label="ORF\frame\1" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 CDS 6401..2124 /label="hSpCas9" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e misc_feature 6482..6496 /label="NLS" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd rep_origin 2603..2909 /label="f1_origin" /ApEinfo_revcolor=#c7b0e3 /ApEinfo_fwdcolor=#c7b0e3 misc_feature 6425..6445 /label="FLAG" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc misc_feature complement(2149..2166) /label="BGH_rev_primer" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 6404..6424 /label="FLAG" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac misc_feature 6446..6469 /label="FLAG" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature 5305..5324 /label="LKO1_5_primer" /ApEinfo_revcolor=#75c6a9 /ApEinfo_fwdcolor=#75c6a9 misc_feature 5385..5398 /label="insert\guide\here" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc rep_origin 4443..5062 /label="pBR322_origin" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 terminator 2152..2355 /label="bGH_PA_terminator" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 misc_feature 5594..5863 /label="CAG_enhancer" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 gene 3428..4288 /label="Ampicillin gene" /ApEinfo_revcolor=#b7e6d7 /ApEinfo_fwdcolor=#b7e6d7 promoter 5143..5375 /label="hU6_promoter" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 promoter 3358..3386 /label="AmpR_promoter" /ApEinfo_revcolor=#f58a5e /ApEinfo_fwdcolor=#f58a5e CDS complement(6380..489) /label="ORF\frame\3" /ApEinfo_revcolor=#c7b0e3 /ApEinfo_fwdcolor=#c7b0e3 misc_feature 6476..6496 /label="SV40\NLS" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 misc_feature 6476..6499 /label="SV40\NLS" /ApEinfo_revcolor=#9eafd2 /ApEinfo_fwdcolor=#9eafd2 misc_feature 5477..5482 /label="U6\terminator" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 misc_feature 3177..3199 /label="pGEX_3_primer" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 5401..5476 /label="Guide\RNA\Scaffold" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 ORIGIN 1 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc 61 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc 121 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc 181 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag 241 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc 301 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa 361 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag 421 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat 481 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg 541 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag 601 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc 661 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc 721 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc 781 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc 841 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag 901 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg 961 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac 1021 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc 1081 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc 1141 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc 1201 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag 1261 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat 1321 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc 1381 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg 1441 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat 1501 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag 1561 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc 1621 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac 1681 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg 1741 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc 1801 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac 1861 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat 1921 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc 1981 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag 2041 acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc 2101 ggccaggcaa aaaagaaaaa gtaagaattc ctagagctcg ctgatcagcc tcgactgtgc 2161 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 2221 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 2281 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 2341 agaatagcag gcatgctggg gagcggccgc aggaacccct agtgatggag ttggccactc 2401 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg 2461 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga 2521 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc 2581 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 2641 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 2701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 2761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag 2821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 2881 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga 2941 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 3001 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac 3061 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc 3121 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 3181 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct 3241 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg 3301 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 3361 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag 3421 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg 3481 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt 3541 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt 3601 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt 3661 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa 3721 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag 3781 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac 3841 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac 3901 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac 3961 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac 4021 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact 4081 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg 4141 tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt 4201 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat 4261 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta 4321 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 4381 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 4441 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 4501 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 4561 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 4621 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 4681 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 4741 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 4801 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 4861 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 4921 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 4981 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 5041 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 5101 tcacatgtcc ttaattaacc ttcgaaccca tgtgagggcc tatttcccat gattccttca 5161 tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac 5221 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca 5281 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc 5341 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct 5401 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt 5461 ggcaccgagt cggtgctttt ttgttttaga gctagaaata gcaagttaaa ataaggctag 5521 tccgttttta gcgcgtgcgc caattctgca gacaaatggc tctagggtta attaaggtct 5581 agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 5641 ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga 5701 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 5761 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt 5821 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga 5881 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt 5941 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg 6001 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc 6061 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg 6121 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc tgccttcgcc 6181 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac 6241 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga 6301 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc 6361 tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata aggaccacga 6421 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa 6481 gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg 6541 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc 6601 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat 6661 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc 6721 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa 6781 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga 6841 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta 6901 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa 6961 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt 7021 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct 7081 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc 7141 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca 7201 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct 7261 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa 7321 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga 7381 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt 7441 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca 7501 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa 7561 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag 7621 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga 7681 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg 7741 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga 7801 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat 7861 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa 7921 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc 7981 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct 8041 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa 8101 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat 8161 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta 8221 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa 8281 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga 8341 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga 8401 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat 8461 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa 8521 cggcatccgg gacaagcagt ccggcaag //