LOCUS       p60                     9688 bp    DNA     circular     22-AUG-2017
DEFINITION  .
ACCESSION   urn.local...9e-7o8q8b3
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     primer          63..86
                     /label="Sequencing XhoI Rev primer"
     misc_feature    179..973
                     /label="Kan R"
     primer          426..445
                     /label="TDNA-pU 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"
     primer          1901..1924
                     /label="P21"
     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    2037..2044
                     /Recognition_pattern="GTTT^AAAC"
                     /label="PmeI"
                     /note="Geneious type: restriction site"
     LTR             complement(2045..3160)
                     /label="NTPII"
     primer          2055..2074
                     /label="NPTII-F"
     misc_feature    2582..2587
                     /Recognition_pattern="C^CATGG"
                     /label="NcoI"
                     /note="Geneious type: restriction site"
     misc_feature    2965..2970
                     /Recognition_pattern="CTGCA^G"
                     /label="PstI"
                     /note="Geneious type: restriction site"
     primer          3135..3153
                     /label="NPTII-R"
     misc_feature    3161..3166
                     /Recognition_pattern="G^GTACC"
                     /label="Acc65I"
                     /note="Geneious type: restriction site"
     misc_feature    complement(3167..3997)
                     /label="35 S promoter"
     misc_feature    3998..4003
                     /Recognition_pattern="G^AATTC"
                     /label="EcoRI"
                     /note="Geneious type: restriction site"
     allele          4004..4308
                     /label="AtU6"
     misc_feature    4194..4199
                     /Recognition_pattern="AGG^CCT"
                     /label="StuI"
                     /note="Geneious type: restriction site"
     attenuator      4313..4319
                     /label="re-AarI"
     attenuator      4330..4336
                     /label="AarI"
     S_region        4341..4423
                     /label="sgRNA scaffold"
     misc_feature    4424..4429
                     /label="Barcode"
     misc_feature    4430..4435
                     /Recognition_pattern="A^AGCTT"
                     /label="HindIII"
                     /note="Geneious type: restriction site"
     -35_signal      4464..5244
                     /label="35S promoter"
     misc_feature    5277..5282
                     /Recognition_pattern="G^CTAGC"
                     /label="NheI"
                     /note="Geneious type: restriction site"
     allele          5283..9420
                     /label="At-Cas9"
     misc_feature    7471..7476
                     /Recognition_pattern="G^GATCC"
                     /label="BamHI"
                     /note="Geneious type: restriction site"
     insertion       7472..9612
                     /label="Insert 1"
     misc_feature    9409..9414
                     /Recognition_pattern="A^GATCT"
                     /label="BglII"
                     /note="Geneious type: restriction site"
     misc_feature    9421..9426
                     /Recognition_pattern="C^CCGGG"
                     /label="XmaI"
                     /note="Geneious type: restriction site"
     misc_feature    9610..9615
                     /Recognition_pattern="AGG^CCT"
                     /label="StuI"
                     /note="Geneious type: restriction site"
     misc_feature    9616..9639
                     /label="T-DNA RB"
     misc_feature    9683..9688
                     /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 aaacagtgtt ttactcctca tattaacttc ggtcattaga ggccacgatt tgacacattt
     2101 ttactcaaaa caaaatgttt gcatatctct tataatttca aattcaacac acaacaaata
     2161 agagaaaaaa caaataatat taatttgaga atgaacaaaa ggaccatatc attcattaac
     2221 tcttctccat ccatttccat ttcacagttc gatagcgaaa accgaataaa aaacacagta
     2281 aattacaagc acaacaaatg gtacaagaaa aacagttttc ccaatgccat aatactcgaa
     2341 cgtccggagt tatcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     2401 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     2461 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     2521 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     2581 gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag
     2641 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     2701 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     2761 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     2821 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     2881 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     2941 ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt
     3001 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     3061 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     3121 tgcgtgcaat ccatcttgtt caatccaagc tcccattgtt ggtacccagc ttgggtctag
     3181 tcgtattaag agatagattt gtagagagag actggtgatt tcagcgtgtc ctctccaaat
     3241 gaaatgaact tccttatata gaggaaggtc ttgcgaagga tagtgggatt gtgcgtcatc
     3301 ccttacgtca gtggagatat cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc
     3361 ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga
     3421 ggcatcttga acgatagcct ttcctttatc gcaatgatgg catttgtagg tgccaccttc
     3481 cttttctact gtccttttga tgaagtgaca gatagctggg caatggaatc cgaggaggtt
     3541 tcccgatatt accctttgtt gaaaagtctc aatagccctt tggtcttctg agactgtatc
     3601 tttgatattc ttggagtaga cgagagtgtc gtgctccacc atgttatcac atcaatccac
     3661 ttgctttgaa gacgtggttg gaacgtcttc tttttccacg atgctcctcg tgggtggggg
     3721 tccatctttg ggaccactgt cggcagaggc atcttgaacg atagcctttc ctttatcgca
     3781 atgatggcat ttgtaggtgc caccttcctt ttctactgtc cttttgatga agtgacagat
     3841 agctgggcaa tggaatccga ggaggtttcc cgatattacc ctttgttgaa aagtctcaat
     3901 agccctttgg tcttctgaga ctgtatcttt gatattcttg gagtagacga gagtgtcgtg
     3961 ctccaccatt acataggccc atcggagcta acgcagtgaa ttcagaaatc tcaaaattcc
     4021 ggcagaacaa ttttgaatct cgatccgtag aaacgagacg gtcattgttt tagttccacc
     4081 acgattatat ttgaaattta cgtgagtgtg agtgagactt gcataagaaa ataaaatctt
     4141 tagttgggaa aaaattcaat aatataaatg ggcttgagaa ggaagcgagg gataggcctt
     4201 tttctaaaat aggcccattt aagctattaa caatcttcaa aagtaccaca gcgcttaggt
     4261 aaagaaagca gctgagttta tatatggtta gagacgaagt agtgattgga tggcaggtgg
     4321 aagaatggac acctgcgaga gttttagagc tagaaatagc aagttaaaat aaggctagtc
     4381 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttagttcca agcttactgc
     4441 gttagctccg atgggcctat gtaatggtgg agcacgacac tctcgtctac tccaagaata
     4501 tcaaagatac agtctcagaa gaccaaaggg ctattgagac ttttcaacaa agggtaatat
     4561 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag
     4621 aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag
     4681 atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
     4741 aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgataac atggtggagc
     4801 acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac caaagggcta
     4861 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     4921 tctgtcactt catcaaaagg acagtagaaa aggaaggtgg cacctacaaa tgccatcatt
     4981 gcgataaagg aaaggctatc gttcaagatg cctctgccga cagtggtccc aaagatggac
     5041 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     5101 tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
     5161 aagaccttcc tctatataag gaagttcatt tcatttggag aggacacgct gaaatcacca
     5221 gtctctctct acaaatctat ctcttaatac gactcactat agggagaccc aagctggcta
     5281 gcaacaatgg ataagaagta ctctatcgga ctcgatatcg gaactaactc tgttggatgg
     5341 gctgtgatca ccgatgagta caaggtgcca tctaagaagt tcaaggttct cggaaacacc
     5401 gataggcact ctatcaagaa aaaccttatc ggtgctctcc tcttcgattc tggtgaaact
     5461 gctgaggcta ccagactcaa gagaaccgct agaagaaggt acaccagaag aaagaacagg
     5521 atctgctacc tccaagagat tttctctaac gagatggcta aagtggatga ttcattcttc
     5581 cacaggctcg aagagtcatt cctcgtggaa gaagataaga agcacgagag gcaccctatc
     5641 ttcggaaaca tcgttgatga ggtggcatac cacgagaagt accctactat ctaccacctc
     5701 agaaagaagc tcgttgattc tactgataag gctgatctca ggctcatcta cctcgctctc
     5761 gctcacatga tcaagttcag aggacacttc ctcatcgagg gtgatctcaa ccctgataac
     5821 tctgatgtgg ataagttgtt catccagctc gtgcagacct acaaccagct tttcgaagag
     5881 aaccctatca acgcttcagg tgtggatgct aaggctatcc tctctgctag gctctctaag
     5941 tcaagaaggc ttgagaacct cattgctcag ctccctggtg agaagaagaa cggacttttc
     6001 ggaaacttga tcgctctctc tctcggactc acccctaact tcaagtctaa cttcgatctc
     6061 gctgaggatg caaagctcca gctctcaaag gatacctacg atgatgatct cgataacctc
     6121 ctcgctcaga tcggagatca gtacgctgat ttgttcctcg ctgctaagaa cctctctgat
     6181 gctatcctcc tcagtgatat cctcagggtg aacaccgaga tcaccaaggc tccactttct
     6241 gcttctatga tcaagagata cgatgagcac caccaggatc tcacacttct caaggctctt
     6301 gttagacagc agctcccaga gaagtacaaa gaaatcttct tcgatcagtc taagaacgga
     6361 tacgctggtt acatcgatgg tggtgcatct caagaagagt tctacaagtt catcaagcca
     6421 atcttggaga agatggatgg aaccgaggaa ctcctcgtga agctcaatag agaggatctc
     6481 cttaggaagc agaggacctt cgataacgga tctatccctc atcagatcca cctcggagag
     6541 ttgcacgcta tccttagaag gcaagaggat ttctacccat tcctcaagga taacagagag
     6601 aagattgaga agatcctcac cttcagaatc ccttactacg tgggacctct cgctagagga
     6661 aactcaagat tcgcttggat gaccagaaag tctgaggaaa ccatcacccc ttggaacttc
     6721 gaagaggtgg tggataaggg tgctagtgct cagtctttca tcgagaggat gaccaacttc
     6781 gataagaacc ttcctaacga gaaggtgctc cctaagcact ctttgctcta cgagtacttc
     6841 accgtgtaca acgagttgac caaggttaag tacgtgaccg agggaatgag gaagcctgct
     6901 tttttgtcag gtgagcaaaa gaaggctatc gttgatctct tgttcaagac caacagaaag
     6961 gtgaccgtga agcagctcaa agaggattac ttcaagaaaa tcgagtgctt cgattcagtg
     7021 gaaatctctg gtgttgagga taggttcaac gcatctctcg gaacctacca cgatctcctc
     7081 aagatcatta aggataagga tttcttggat aacgaggaaa acgaggatat cttggaggat
     7141 atcgttctta ccctcaccct cttcgaggat agagagatga tagaagaaag gctcaagacc
     7201 tacgctcatc tcttcgatga taaggtgatg aagcagttga agagaagaag atacactggt
     7261 tggggaaggc tctcaagaaa gctcattaac ggaatcaggg ataagcagtc tggaaagaca
     7321 atccttgatt tcctcaagtc tgatggattc gctaacagaa acttcatgca gctcatccac
     7381 gatgattctc tcacctttaa agaggatatc cagaaggctc aggtttcagg acagggtgat
     7441 agtctccatg agcatatcgc taacctcgct ggatcccctg caatcaagaa gggaatcctc
     7501 cagactgtga agattgtgga tgagttggtg aaggtgatgg gacacaagcc tgagaacatc
     7561 gtgatcgaaa tggctagaga gaaccagacc actcagaagg gacagaagaa ctctagggaa
     7621 aggatgaaga ggatcgagga aggtatcaaa gagcttggat ctcagatcct caaagagcac
     7681 cctgttgaga acactcagct ccagaacgag aagctctacc tctactactt gcagaacgga
     7741 agggatatgt atgtggatca agagcttgat attaacaggc tctctgatta cgatgttgat
     7801 catatcgtgc cacagtcttt tatcaaagat gattctatcg ataacaaggt gctcactagg
     7861 tctgataaga acaggggtaa gagtgataac gtgccaagtg aagaggttgt gaagaaaatg
     7921 aagaactatt ggaggcagct cctcaacgct aagctcatca ctcagagaaa gttcgataac
     7981 ttgaccaagg ctgagagggg aggactctct gaattggata aggcaggatt catcaagaga
     8041 cagctcgtgg aaaccaggca gatcaccaaa catgtggcac agatcctcga ttctaggatg
     8101 aacaccaagt acgatgagaa cgataagttg atcagggaag tgaaggttat caccctcaag
     8161 tcaaagctcg tgtctgattt cagaaaggat ttccaattct acaaggtgag ggaaatcaac
     8221 aactaccacc acgctcacga tgcttacctt aacgctgttg ttggaaccgc tctcatcaag
     8281 aagtatccaa agttggagtc tgagttcgtg tacggtgatt ataaggtgta cgatgtgagg
     8341 aagatgatcg ctaagtctga gcaagagatc ggaaaggcta ccgctaagta tttcttctac
     8401 tctaacatca tgaatttctt caagaccgag atcactctcg ctaacggtga gatcagaaag
     8461 aggccactca tcgagacaaa cggtgaaaca ggtgagatcg tgtgggataa gggaagggat
     8521 ttcgctaccg ttagaaaggt gctctctatg cctcaggtga acatcgttaa gaaaaccgag
     8581 gtgcagaccg gtggattctc taaagagtct atcctcccta agaggaactc tgataagctc
     8641 attgctagga agaaggattg ggaccctaag aaatacggtg gtttcgattc tcctaccgtg
     8701 gcttactctg ttctcgttgt ggctaaggtt gagaagggaa agagtaagaa gctcaagtct
     8761 gttaaggaac ttctcggaat cactatcatg gaaaggtcat ctttcgagaa gaacccaatc
     8821 gatttccttg aggctaaggg atacaaagag gttaagaagg atctcatcat caagctccca
     8881 aagtactcac ttttcgagtt ggagaacggt agaaagagga tgctcgcttc tgctggtgag
     8941 cttcaaaagg gaaacgagct tgctctccca tctaagtacg ttaactttct ttacctcgct
     9001 tctcactacg agaagttgaa gggatctcca gaagataacg agcagaagca acttttcgtt
     9061 gagcagcaca agcactactt ggatgagatc atcgagcaga tcagtgagtt ctctaaaagg
     9121 gtgatcctcg ctgatgcaaa cctcgataag gtgttgtctg cttacaacaa gcacagagat
     9181 aagcctatca gggaacaggc agagaacatc atccatctct tcacccttac caacctcggt
     9241 gctcctgctg ctttcaagta cttcgataca accatcgata ggaagagata cacctctacc
     9301 aaagaagtgc tcgatgctac cctcatccat cagtctatca ctggactcta cgagactagg
     9361 atcgatctct cacagcttgg aggtgatcct aagaagaaaa gaaaggttag atcttgatga
     9421 cccgggtctc cataataatg tgtgagtagt tcccagataa gggaattagg gttcctatag
     9481 ggtttcgctc atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat
     9541 acttctatca ataaaatttc taattcctaa aaccaaaatc cagtactaaa atccagatcc
     9601 cccgaattaa ggccttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta
     9661 ttagaataac ggatatttaa aactcgag
//
