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
//