LOCUS       p59                     9874 bp    DNA     circular     22-AUG-2017
DEFINITION  .
ACCESSION   urn.local...99-7o8q7zu
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     primer          63..86
                     /label="Sequencing XhoI Rev primer"
     misc_feature    179..973
                     /label="Kan R"
     primer_bind     426..445
                     /label="pUC-THMS2 primer Rev"
     misc_feature    739..744
                     /Recognition_pattern="C^CATGG"
                     /label="NcoI"
                     /note="Geneious type: restriction site"
     misc_feature    1273..1946
                     /label="pUC Ori"
     primer_bind     1823..1842
                     /label="pUC-THMS2 primer Forw"
     misc_feature    1981..1986
                     /Recognition_pattern="G^TCGAC"
                     /label="SalI"
                     /note="Geneious type: restriction site"
     misc_feature    2005..2010
                     /Recognition_pattern="T^CTAGA"
                     /label="XbaI"
                     /note="Geneious type: restriction site"
     misc_feature    2011..2036
                     /label="T-DNA LB"
     misc_feature    complement(2037..2312)
                     /label="NOS terminator"
     misc_feature    complement(2313..3345)
                     /label="Hygromycin R"
     primer_bind     2373..2392
                     /label="pUC-THMS2 primer Rev"
     misc_feature    2984..2989
                     /Recognition_pattern="C^CATGG"
                     /label="NcoI"
                     /note="Geneious type: restriction site"
     misc_feature    3346..3351
                     /Recognition_pattern="G^GTACC"
                     /label="Acc65I"
                     /note="Geneious type: restriction site"
     misc_feature    3346..3351
                     /Recognition_pattern="GGTAC^C"
                     /label="KpnI"
                     /note="Geneious type: restriction site"
     misc_feature    complement(3352..4182)
                     /label="35 S promoter"
     misc_feature    4183..4188
                     /Recognition_pattern="G^AATTC"
                     /label="EcoRI"
                     /note="Geneious type: restriction site"
     allele          4189..4497
                     /label="atU6"
     misc_feature    4379..4384
                     /Recognition_pattern="AGG^CCT"
                     /label="StuI"
                     /note="Geneious type: restriction site"
     S_region        4526..4608
                     /label="sgRNA scaffold"
     misc_feature    4609..4614
                     /label="Barcode"
     misc_feature    4616..4621
                     /Recognition_pattern="A^AGCTT"
                     /label="HindIII"
                     /note="Geneious type: restriction site"
     -35_signal      4650..5430
                     /label="35S promoter"
     misc_feature    5463..5468
                     /Recognition_pattern="G^CTAGC"
                     /label="NheI"
                     /note="Geneious type: restriction site"
     C_region        5473..9603
                     /label="AtCas9"
     misc_feature    7657..7662
                     /Recognition_pattern="G^GATCC"
                     /label="BamHI"
                     /note="Geneious type: restriction site"
     misc_feature    9595..9600
                     /Recognition_pattern="A^GATCT"
                     /label="BglII"
                     /note="Geneious type: restriction site"
     misc_feature    9607..9612
                     /Recognition_pattern="C^CCGGG"
                     /label="XmaI"
                     /note="Geneious type: restriction site"
     misc_feature    9796..9801
                     /Recognition_pattern="AGG^CCT"
                     /label="StuI"
                     /note="Geneious type: restriction site"
     misc_feature    9802..9825
                     /label="T-DNA RB"
     misc_feature    9869..9874
                     /label="XhoI"
ORIGIN      
        1 ctcgagcttc tactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc
       61 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct cgccgccaag
      121 gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat cgtttcgcat
      181 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
      241 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
      301 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
      361 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
      421 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
      481 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
      541 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
      601 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
      661 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
      721 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
      781 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
      841 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
      901 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
      961 cgagttcttc tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     1021 tctgtgcggt atttcacacc gcatacaggt ggcacttttc ggggaaatgt gcgcggaacc
     1081 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1141 tgataaatgc ttcaataata gcacgtgcta aaacttcatt tttaatttaa aaggatctag
     1201 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1261 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1381 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1441 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1501 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1561 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1621 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1681 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1741 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1801 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1861 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1921 ggcttttgct ggccttttgc tcacatgttc ttgactcttc gcgatgtacg ggccagatat
     1981 gtcgacgtaa aacgacggcc agtgtctaga tggcaggata tattgtggtg taaacagttt
     2041 aaacccgatc tagtaacata gatagcaccg cgcgcgataa tttatcctag tttgcgcgct
     2101 atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc
     2161 atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca
     2221 gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt
     2281 gccaaatgtt tgaacgatcg gggagtttaa acttactatt cctttgccct cggacgagtg
     2341 ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt ccagacggcc
     2401 gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg gacgattgcg
     2461 tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa gctctgatag
     2521 agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc tgcaagctcc
     2581 ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca cggcctccag
     2641 aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg
     2701 accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg
     2761 aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag agcctgcgcg
     2821 acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg gggatcagca
     2881 atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg tccgaatggg
     2941 ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc ctccgcgacc
     3001 ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac accctgtgca
     3061 cggcgggaga tgcaataggt caggctctcg ctgaactccc caatgtcaag cacttccgga
     3121 atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta gaaaccatcg
     3181 gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct gaaagcacga
     3241 gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga
     3301 aacttctcga cagacgtcgc ggtgagttca ggctttttca ttgttggtac ccagcttggg
     3361 tctagtcgta ttaagagata gatttgtaga gagagactgg tgatttcagc gtgtcctctc
     3421 caaatgaaat gaacttcctt atatagagga aggtcttgcg aaggatagtg ggattgtgcg
     3481 tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga
     3541 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
     3601 gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt gtaggtgcca
     3661 ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg gaatccgagg
     3721 aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
     3781 gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt atcacatcaa
     3841 tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt
     3901 gggggtccat ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta
     3961 tcgcaatgat ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga
     4021 cagatagctg ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc
     4081 tcaatagccc tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg
     4141 tcgtgctcca ccattacata ggcccatcgg agctaacgca gtgaattcag aaatctcaaa
     4201 attccggcag aacaattttg aatctcgatc cgtagaaacg agacggtcat tgttttagtt
     4261 ccaccacgat tatatttgaa atttacgtga gtgtgagtga gacttgcata agaaaataaa
     4321 atctttagtt gggaaaaaat tcaataatat aaatgggctt gagaaggaag cgagggatag
     4381 gcctttttct aaaataggcc catttaagct attaacaatc ttcaaaagta ccacagcgct
     4441 taggtaaaga aagcagctga gtttatatat ggttagagac gaagtagtga ttggatggca
     4501 ggtggaagaa tggacacctg cgagagtttt agagctagaa atagcaagtt aaaataaggc
     4561 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttgg ctacaaagct
     4621 tactgcgtta gctccgatgg gcctatgtaa tggtggagca cgacactctc gtctactcca
     4681 agaatatcaa agatacagtc tcagaagacc aaagggctat tgagactttt caacaaaggg
     4741 taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcaaaagga
     4801 cagtagaaaa ggaaggtggc acctacaaat gccatcattg cgataaagga aaggctatcg
     4861 ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
     4921 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gataacatgg
     4981 tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa
     5041 gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc
     5101 cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc
     5161 atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt ggtcccaaag
     5221 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
     5281 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
     5341 cttcgcaaga ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctgaaa
     5401 tcaccagtct ctctctacaa atctatctct taatacgact cactataggg agacccaagc
     5461 tggctagcaa caatggataa gaagtactct atcggactcg atatcggaac taactctgtt
     5521 ggatgggctg tgatcaccga tgagtacaag gtgccatcta agaagttcaa ggttctcgga
     5581 aacaccgata ggcactctat caagaaaaac cttatcggtg ctctcctctt cgattctggt
     5641 gaaactgctg aggctaccag actcaagaga accgctagaa gaaggtacac cagaagaaag
     5701 aacaggatct gctacctcca agagattttc tctaacgaga tggctaaagt ggatgattca
     5761 ttcttccaca ggctcgaaga gtcattcctc gtggaagaag ataagaagca cgagaggcac
     5821 cctatcttcg gaaacatcgt tgatgaggtg gcataccacg agaagtaccc tactatctac
     5881 cacctcagaa agaagctcgt tgattctact gataaggctg atctcaggct catctacctc
     5941 gctctcgctc acatgatcaa gttcagagga cacttcctca tcgagggtga tctcaaccct
     6001 gataactctg atgtggataa gttgttcatc cagctcgtgc agacctacaa ccagcttttc
     6061 gaagagaacc ctatcaacgc ttcaggtgtg gatgctaagg ctatcctctc tgctaggctc
     6121 tctaagtcaa gaaggcttga gaacctcatt gctcagctcc ctggtgagaa gaagaacgga
     6181 cttttcggaa acttgatcgc tctctctctc ggactcaccc ctaacttcaa gtctaacttc
     6241 gatctcgctg aggatgcaaa gctccagctc tcaaaggata cctacgatga tgatctcgat
     6301 aacctcctcg ctcagatcgg agatcagtac gctgatttgt tcctcgctgc taagaacctc
     6361 tctgatgcta tcctcctcag tgatatcctc agggtgaaca ccgagatcac caaggctcca
     6421 ctttctgctt ctatgatcaa gagatacgat gagcaccacc aggatctcac acttctcaag
     6481 gctcttgtta gacagcagct cccagagaag tacaaagaaa tcttcttcga tcagtctaag
     6541 aacggatacg ctggttacat cgatggtggt gcatctcaag aagagttcta caagttcatc
     6601 aagccaatct tggagaagat ggatggaacc gaggaactcc tcgtgaagct caatagagag
     6661 gatctcctta ggaagcagag gaccttcgat aacggatcta tccctcatca gatccacctc
     6721 ggagagttgc acgctatcct tagaaggcaa gaggatttct acccattcct caaggataac
     6781 agagagaaga ttgagaagat cctcaccttc agaatccctt actacgtggg acctctcgct
     6841 agaggaaact caagattcgc ttggatgacc agaaagtctg aggaaaccat caccccttgg
     6901 aacttcgaag aggtggtgga taagggtgct agtgctcagt ctttcatcga gaggatgacc
     6961 aacttcgata agaaccttcc taacgagaag gtgctcccta agcactcttt gctctacgag
     7021 tacttcaccg tgtacaacga gttgaccaag gttaagtacg tgaccgaggg aatgaggaag
     7081 cctgcttttt tgtcaggtga gcaaaagaag gctatcgttg atctcttgtt caagaccaac
     7141 agaaaggtga ccgtgaagca gctcaaagag gattacttca agaaaatcga gtgcttcgat
     7201 tcagtggaaa tctctggtgt tgaggatagg ttcaacgcat ctctcggaac ctaccacgat
     7261 ctcctcaaga tcattaagga taaggatttc ttggataacg aggaaaacga ggatatcttg
     7321 gaggatatcg ttcttaccct caccctcttc gaggatagag agatgataga agaaaggctc
     7381 aagacctacg ctcatctctt cgatgataag gtgatgaagc agttgaagag aagaagatac
     7441 actggttggg gaaggctctc aagaaagctc attaacggaa tcagggataa gcagtctgga
     7501 aagacaatcc ttgatttcct caagtctgat ggattcgcta acagaaactt catgcagctc
     7561 atccacgatg attctctcac ctttaaagag gatatccaga aggctcaggt ttcaggacag
     7621 ggtgatagtc tccatgagca tatcgctaac ctcgctggat cccctgcaat caagaaggga
     7681 atcctccaga ctgtgaagat tgtggatgag ttggtgaagg tgatgggaca caagcctgag
     7741 aacatcgtga tcgaaatggc tagagagaac cagaccactc agaagggaca gaagaactct
     7801 agggaaagga tgaagaggat cgaggaaggt atcaaagagc ttggatctca gatcctcaaa
     7861 gagcaccctg ttgagaacac tcagctccag aacgagaagc tctacctcta ctacttgcag
     7921 aacggaaggg atatgtatgt ggatcaagag cttgatatta acaggctctc tgattacgat
     7981 gttgatcata tcgtgccaca gtcttttatc aaagatgatt ctatcgataa caaggtgctc
     8041 actaggtctg ataagaacag gggtaagagt gataacgtgc caagtgaaga ggttgtgaag
     8101 aaaatgaaga actattggag gcagctcctc aacgctaagc tcatcactca gagaaagttc
     8161 gataacttga ccaaggctga gaggggagga ctctctgaat tggataaggc aggattcatc
     8221 aagagacagc tcgtggaaac caggcagatc accaaacatg tggcacagat cctcgattct
     8281 aggatgaaca ccaagtacga tgagaacgat aagttgatca gggaagtgaa ggttatcacc
     8341 ctcaagtcaa agctcgtgtc tgatttcaga aaggatttcc aattctacaa ggtgagggaa
     8401 atcaacaact accaccacgc tcacgatgct taccttaacg ctgttgttgg aaccgctctc
     8461 atcaagaagt atccaaagtt ggagtctgag ttcgtgtacg gtgattataa ggtgtacgat
     8521 gtgaggaaga tgatcgctaa gtctgagcaa gagatcggaa aggctaccgc taagtatttc
     8581 ttctactcta acatcatgaa tttcttcaag accgagatca ctctcgctaa cggtgagatc
     8641 agaaagaggc cactcatcga gacaaacggt gaaacaggtg agatcgtgtg ggataaggga
     8701 agggatttcg ctaccgttag aaaggtgctc tctatgcctc aggtgaacat cgttaagaaa
     8761 accgaggtgc agaccggtgg attctctaaa gagtctatcc tccctaagag gaactctgat
     8821 aagctcattg ctaggaagaa ggattgggac cctaagaaat acggtggttt cgattctcct
     8881 accgtggctt actctgttct cgttgtggct aaggttgaga agggaaagag taagaagctc
     8941 aagtctgtta aggaacttct cggaatcact atcatggaaa ggtcatcttt cgagaagaac
     9001 ccaatcgatt tccttgaggc taagggatac aaagaggtta agaaggatct catcatcaag
     9061 ctcccaaagt actcactttt cgagttggag aacggtagaa agaggatgct cgcttctgct
     9121 ggtgagcttc aaaagggaaa cgagcttgct ctcccatcta agtacgttaa ctttctttac
     9181 ctcgcttctc actacgagaa gttgaaggga tctccagaag ataacgagca gaagcaactt
     9241 ttcgttgagc agcacaagca ctacttggat gagatcatcg agcagatcag tgagttctct
     9301 aaaagggtga tcctcgctga tgcaaacctc gataaggtgt tgtctgctta caacaagcac
     9361 agagataagc ctatcaggga acaggcagag aacatcatcc atctcttcac ccttaccaac
     9421 ctcggtgctc ctgctgcttt caagtacttc gatacaacca tcgataggaa gagatacacc
     9481 tctaccaaag aagtgctcga tgctaccctc atccatcagt ctatcactgg actctacgag
     9541 actaggatcg atctctcaca gcttggaggt gatcctaaga agaaaagaaa ggttagatct
     9601 tgatgacccg ggtctccata ataatgtgtg agtagttccc agataaggga attagggttc
     9661 ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
     9721 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc
     9781 agatcccccg aattaaggcc ttgacaggat atattggcgg gtaaacctaa gagaaaagag
     9841 cgtttattag aataacggat atttaaaact cgag
//
