LOCUS       sgRNA_RAF1_94          10394 bp ds-DNA     circular     12-OCT-2017
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             6247..6957
                     /label="mCherry"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     promoter        297..314
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     enhancer        1000..1379
                     /label="CMV enhancer"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     protein_bind    8711..8727
                     /label="lac operator"
                     /ApEinfo_revcolor=#31849b
                     /ApEinfo_fwdcolor=#31849b
     protein_bind    255..276
                     /label="CAP binding site"
                     /ApEinfo_revcolor=#31849b
                     /ApEinfo_fwdcolor=#31849b
     CDS             1666..5769
                     /label="Cas9"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     rep_origin      6034..6169
                     /label="SV40 ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    763..963
                     /label="g-block"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             5785..5805
                     /label="SV40 NLS"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     primer_bind     353..369
                     /label="M13 rev"
                     /ApEinfo_revcolor=#a020f0
                     /ApEinfo_fwdcolor=#a020f0
     promoter        315..321
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    816..836
                     /label="Seed Sequence"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     promoter        291..296
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        1380..1583
                     /label="CMV promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        389..407
                     /label="SP6 promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        1628..1646
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer_bind     complement(8687..8703)
                     /label="M13 rev"
                     /ApEinfo_revcolor=#a020f0
                     /ApEinfo_fwdcolor=#a020f0
     CDS             7582..8385
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     rep_origin      9796..10384
                     /label="ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     protein_bind    329..345
                     /label="lac operator"
                     /ApEinfo_revcolor=#31849b
                     /ApEinfo_fwdcolor=#31849b
     CDS             8789..8857
                     /label="AmpR"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     promoter        419..437
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    6973..7546
                     /label="IRES"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     polyA_signal    8517..8638
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     promoter        5854..6183
                     /label="SV40 promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             8858..9625
                     /label="AmpR"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             5806..5832
                     /label="HA"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
ORIGIN
        1 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
       61 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      121 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      181 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      241 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      301 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      361 agctatgaca tgattacgaa ttgcaacgat ttaggtgaca ctatagaaga gaaggaatta
      421 atacgactca ctatagggag agagagagaa ttaccctcac taaagggagg agaagcatga
      481 attccccagt ggaaagacgc gcaggcaaaa cgcaccacgt gacggagcgt gaccgcgcgc
      541 cgagcgcgcg ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg
      601 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
      661 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      721 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa ctTGAAAGTA TTTCGATTTC
      781 TTGGGTTTAT ATATCTTGTG GAAAGGACGA GGATCGCCGC CCGAGAGTCT TAATCGGTTT
      841 TAGAGCTAGA AATAGCAAGT TAAAATAAGG CTAGTCCGTT ATCAACTTGA AAAAGTGGCA
      901 CCGAGTCGGT GCTTTTTTCT AGACACAATT GCATGAAGAA TCTGCTTAGG GTTAGGCGTT
      961 TTGcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     1021 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1081 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1141 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1201 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1261 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1321 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1381 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1441 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1501 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1561 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     1621 tcgaaattaa tacgactcac tatagggaga cccaagcttg ccaccatgga caagaagtac
     1681 agcatcggcc tggacatcgg taccaacagc gtgggctggg ccgtgatcac cgacgagtac
     1741 aaggtgccca gcaagaagtt caaggtgctg ggcaacaccg accgccacag catcaagaag
     1801 aacctgatcg gcgccctgct gttcgacagc ggcgagaccg ccgaggccac ccgcctgaag
     1861 cgcaccgccc gccgccgcta cacccgccgc aagaaccgca tctgctacct gcaggagatc
     1921 ttcagcaacg agatggccaa ggtggacgac agcttcttcc accgcctgga ggagagcttc
     1981 ctggtggagg aggacaagaa gcacgagcgc caccccatct tcggcaacat cgtggacgag
     2041 gtggcctacc acgagaagta ccccaccatc taccacctgc gcaagaagct ggtggacagc
     2101 accgacaagg ccgacctgcg cctgatctac ctggccctgg cccacatgat caagttccgc
     2161 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     2221 atccagctgg tgcagaccta caaccagctg ttcgaggaga accccatcaa cgccagcggc
     2281 gtggacgcca aggccatcct gagcgcccgc ctgagcaaga gccgccgcct ggagaacctg
     2341 atcgcccagc tgcccggcga gaagaagaac ggcctgttcg gcaacctgat cgccctgagc
     2401 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggacgc caagctgcag
     2461 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     2521 tacgccgacc tgttcctggc cgccaagaac ctgagcgacg ccatcctgct gagcgacatc
     2581 ctgcgcgtga acaccgagat caccaaggcc cccctgagcg ccagcatgat caagcgctac
     2641 gacgagcacc accaggacct gaccctgctg aaggccctgg tgcgccagca gctgcccgag
     2701 aagtacaagg agatcttctt cgaccagagc aagaacggct acgccggcta catcgacggc
     2761 ggcgccagcc aggaggagtt ctacaagttc atcaagccca tcctggagaa gatggacggc
     2821 accgaggagc tgctggtgaa gctgaaccgc gaggacctgc tgcgcaagca gcgcaccttc
     2881 gacaacggca gcatccccca ccagatccac ctgggcgagc tgcacgccat cctgcgccgc
     2941 caggaggact tctacccctt cctgaaggac aaccgcgaga agatcgagaa gatcctgacc
     3001 ttccgcatcc cctactacgt gggccccctg gcccgcggca acagccgctt cgcctggatg
     3061 acccgcaaga gcgaggagac catcaccccc tggaacttcg aggaggtggt ggacaagggc
     3121 gccagcgccc agagcttcat cgagcgcatg accaacttcg acaagaacct gcccaacgag
     3181 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc
     3241 aaggtgaagt acgtgaccga gggcatgcgc aagcccgcct tcctgagcgg cgagcagaag
     3301 aaggccatcg tggacctgct gttcaagacc aaccgcaagg tgaccgtgaa gcagctgaag
     3361 gaggactact tcaagaagat cgagtgcttc gacagcgtgg agatcagcgg cgtggaggac
     3421 cgcttcaacg ccagcctggg cacctaccac gacctgctga agatcatcaa ggacaaggac
     3481 ttcctggaca acgaggagaa cgaggacatc ctggaggaca tcgtgctgac cctgaccctg
     3541 ttcgaggacc gcgagatgat cgaggagcgc ctgaagacct acgcccacct gttcgacgac
     3601 aaggtgatga agcagctgaa gcgccgccgc tacaccggct ggggccgcct gagccgcaag
     3661 cttatcaacg gcatccgcga caagcagagc ggcaagacca tcctggactt cctgaagagc
     3721 gacggcttcg ccaaccgcaa cttcatgcag ctgatccacg acgacagcct gaccttcaag
     3781 gaggacatcc agaaggccca ggtgagcggc cagggcgaca gcctgcacga gcacatcgcc
     3841 aacctggccg gcagccccgc catcaagaag ggcatcctgc agaccgtgaa ggtggtggac
     3901 gagctggtga aggtgatggg ccgccacaag cccgagaaca tcgtgatcga gatggcccgc
     3961 gagaaccaga ccacccagaa gggccagaag aacagccgcg agcgcatgaa gcgcatcgag
     4021 gagggcatca aggagctggg cagccagatc ctgaaggagc accccgtgga gaacacccag
     4081 ctgcagaacg agaagctgta cctgtactac ctgcagaacg gccgcgacat gtacgtggac
     4141 caggagctgg acatcaaccg cctgagcgac tacgacgtgg accacatcgt gccccagagc
     4201 ttcctgaagg acgacagcat cgacaacaag gtgctgaccc gcagcgacaa gaaccgcggc
     4261 aagagcgaca acgtgcccag cgaggaggtg gtgaagaaga tgaagaacta ctggcgccag
     4321 ctgctgaacg ccaagctgat cacccagcgc aagttcgaca acctgaccaa ggccgagcgc
     4381 ggcggcctga gcgagctgga caaggccggc ttcatcaagc gccagctggt ggagacccgc
     4441 cagatcacca agcacgtggc ccagatcctg gacagccgca tgaacaccaa gtacgacgag
     4501 aacgacaagc tgatccgcga ggtgaaggtg atcaccctga agagcaagct ggtgagcgac
     4561 ttccgcaagg acttccagtt ctacaaggtg cgcgagatca acaactacca ccacgcccac
     4621 gacgcctacc tgaacgccgt ggtgggcacc gccctgatca agaagtaccc caagctggag
     4681 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc gcaagatgat cgccaagagc
     4741 gagcaggaga tcggcaaggc caccgccaag tacttcttct acagcaacat catgaacttc
     4801 ttcaagaccg agatcaccct ggccaacggc gagatccgca agcgccccct gatcgagacc
     4861 aacggcgaga ccggcgagat cgtgtgggac aagggccgcg acttcgccac cgtgcgcaag
     4921 gtgctgagca tgccccaggt gaacatcgtg aagaagaccg aggtgcagac cggcggcttc
     4981 agcaaggaga gcatcctgcc caagcgcaac agcgacaagc tgatcgcccg caagaaggac
     5041 tgggacccca agaagtacgg cggcttcgac agccccaccg tggcctacag cgtgctggtg
     5101 gtggccaagg tggagaaggg caagagcaag aagctgaaga gcgtgaagga gctgctgggc
     5161 atcaccatca tggagcgcag cagcttcgag aagaacccca tcgacttcct ggaggccaag
     5221 ggctacaagg aggtgaagaa ggacctgatc atcaagctgc ccaagtacag cctgttcgag
     5281 ctggagaacg gccgcaagcg catgctggcc agcgccggcg agctgcagaa gggcaacgag
     5341 ctggccctgc ccagcaagta cgtgaacttc ctgtacctgg ccagccacta cgagaagctg
     5401 aagggcagcc ccgaggacaa cgagcagaag cagctgttcg tggagcagca caagcactac
     5461 ctggacgaga tcatcgagca gatcagcgag ttcagcaagc gcgtgatcct ggccgacgcc
     5521 aacctggaca aggtgctgag cgcctacaac aagcaccgcg acaagcccat ccgcgagcag
     5581 gccgagaaca tcatccacct gttcaccctg accaacctgg gcgcccccgc cgccttcaag
     5641 tacttcgaca ccaccatcga ccgcaagcgc tacaccagca ccaaggaggt gctggacgcc
     5701 accctgatcc accagagcat caccggtctg tacgagaccc gcatcgacct gagccagctg
     5761 ggcggcgacg gcggctccgg acctccaaag aaaaagagaa aagtataccc ctacgacgtg
     5821 cccgactacg cctaataact cgagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     6001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     6061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     6121 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     6181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcggcgcgc cataccggtc
     6241 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     6301 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     6361 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     6421 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     6481 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     6541 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     6601 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     6661 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     6721 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     6781 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     6841 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     6901 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaggcg
     6961 gccgcgatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     7021 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     7081 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     7141 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     7201 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     7261 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     7321 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tcacctcaag
     7381 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     7441 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     7501 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     7561 gcgataattc ctgcagccaa tatgggatcg gccattgaac aagatggatt gcacgcaggt
     7621 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     7681 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     7741 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     7801 gccacgacgg gcgttccttg cgcagcagtg ctcgacgttg tcactgaagc gggaagggac
     7861 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     7921 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     7981 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     8041 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     8101 ttcgccaggc tcaaggcgcg tatgcccgac ggcgatgatc tcgtcgtgac tcatggcgat
     8161 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     8221 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     8281 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     8341 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaacgcg gtgctacgag
     8401 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     8461 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     8521 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     8581 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     8641 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     8701 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagg
     8761 taagcctgaa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     8821 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     8881 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     8941 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     9001 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     9061 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     9121 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     9181 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagggggatc atgtaactcg
     9241 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     9301 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     9361 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     9421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     9481 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     9541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     9601 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     9661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     9721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     9781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     9841 aaaaccaccg ctaccagcgg tggttttttt gccggatcaa gagctaccaa ctctttttcc
     9901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     9961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    10021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    10081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    10141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
    10201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
    10261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
    10321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
    10381 gaaaaacgcc agca
//