LOCUS       sequence-404729-       20438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-KO in Arabidopsis thaliana.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 20438)
  AUTHORS   Ordon J, Kiel N, Becker D, Kretschmer C, Schulze-Lefert P, Stuttmann
            J
  TITLE     Targeted gene deletion with SpCas9 and multiple guide RNAs in 
            Arabidopsis thaliana: four are better than two.
  JOURNAL   Plant Methods. 2023 Mar 28;19(1):30. doi: 
            10.1186/s13007-023-01010-4.
  PUBMED    36978193
REFERENCE   2  (bases 1 to 20438)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Methods."; date: "2023-03-28"; volume: "19(1):30. doi"; pages: " 
            10.1186/s13007-023-01010-4"
COMMENT     Sequence Label: pDGE1108
FEATURES             Location/Qualifiers
     source          1..20438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1062..1691
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2120..3193
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      3259..3453
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3689..3711)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3797..3937
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3852..3871
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3952..3969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4123..4711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4203..4222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4957..5748)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASCADQLEEFVHYVKGEITKVVGK"
     misc_feature    6269..6293
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(6805..7500)
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     CDS             complement(11130..11150)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(16883..16903)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        18613..18715
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             18716..19399
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(18782..18801)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             19719..20024
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     19903..19922
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    20145..20169
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ttcctggcgt tttcttgtcg cgtgttttag tcgcataaag tagaatactt gcgactagaa
       61 ccggagacat tacgccatga acaagagcgc cgccgctggc ctgctgggct atgcccgcgt
      121 cagcaccgac gaccaggact tgaccaacca acgggccgaa ctgcacgcgg ccggctgcac
      181 caagctgttt tccgagaaga tcaccggcac caggcgcgac cgcccggagc tggccaggat
      241 gcttgaccac ctacgccctg gcgacgttgt gacagtgacc aggctagacc gcctggcccg
      301 cagcacccgc gacctactgg acattgccga gagcatccag gaggccggcg cgggcctgcg
      361 tagcctggca gagccgtggg ccgacaccac cacgccggcc ggccgcatgg tgttgaccgt
      421 gttcgccggc attgccgagt tcgagcgttc cctaatcatc gaccgcaccc ggagcgggcg
      481 cgaggccgcc aaggcccgag gcgtgaagtt tggcccccgc cctaccctca ccccggcaca
      541 gatcgcgcac gcccgcgagc tgatcgacca ggaaggccgc accgtgaaag aggcggctgc
      601 actgcttggc gtgcatcgct cgaccctgta ccgcgcactt gagcgcagcg aggaagtgac
      661 gcccaccgag gccaggcggc gcggtgcctt ccgtgaggac gcattgaccg aggccgacgc
      721 cctggcggcc gccgagaatg aacgccaaga ggaacaagca tgaaaccgca ccaggacggc
      781 caggacgaac cgtttttcat taccgaagag atcgaggcgg agatgatcgc ggccgggtac
      841 gtgttcgagc cgcccgcgca cgtctcaacc gtgcggctgc atgaaatcct ggccggtttg
      901 tctgatgcca agctggcggc ctggccggcc agcttggccg ctgaagaaac cgagcgccgc
      961 cgtctaaaaa ggtgatgtgt atttgagtaa aacagcttgc gtcatgcggt cgctgcgtat
     1021 atgatgcgat gagtaaataa acaaatacgc aaggggaacg catgaaggtt atcgctgtac
     1081 ttaaccagaa aggcgggtca ggcaagacga ccatcgcaac ccatctagcc cgcgccctgc
     1141 aactcgccgg ggccgatgtt ctgttagtcg attccgatcc ccagggcagt gcccgcgatt
     1201 gggcggccgt gcgggaagat caaccgctaa ccgttgtcgg catcgaccgc ccgacgattg
     1261 accgcgacgt gaaggccatc ggccggcgcg acttcgtagt gatcgacgga gcgccccagg
     1321 cggcggactt ggctgtgtcc gcgatcaagg cagccgactt cgtgctgatt ccggtgcagc
     1381 caagccctta cgacatatgg gccaccgccg acctggtgga gctggttaag cagcgcattg
     1441 aggtcacgga tggaaggcta caagcggcct ttgtagtgtc gcgggcgatc aaaggcacgc
     1501 gcatcggcgg tgaggttgcc gaggcgctgg ccgggtacga gctgcccatt cttgagtccc
     1561 gtatcacgca gcgcgtgagc tacccaggca ctgccgccgc cggcacaacc gttcttgaat
     1621 cagaacccga gggcgacgct gcccgcgagg tccaggcgct ggccgctgaa attaaatcaa
     1681 aactcatttg agttaatgag gtaaagagaa aatgagcaaa agcacaaaca cgctaagtgc
     1741 cggccgtccg agcgcacgca gcagcaaggc tgcaacgttg gccagcctgg cagacacgcc
     1801 agccatgaag cgggtcaact ttcagttgcc ggcggaggat cacaccaagc tgaagatgta
     1861 cgcggtacgc caaggcaaga ccattaccga gctgctatct gaatacatcg cgcagctacc
     1921 agagtaaatg agcaaatgaa taaatgagta gatgaatttt agcggctaaa ggaggcggca
     1981 tggaaaatca agaacaacca ggcaccgacg ccgtggaatg ccccatgtgt ggaggaacgg
     2041 gcggttggcc aggcgtaagc ggctgggttg tctgccggcc ctgcaatggc actggaaccc
     2101 ccaagcccga ggaatcggcg tgagcggtcg caaaccatcc ggcccggtac aaatcggcgc
     2161 ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg
     2221 catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa
     2281 agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga
     2341 cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag
     2401 catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat
     2461 ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag
     2521 tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg
     2581 ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt
     2641 actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg
     2701 cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg
     2761 cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga
     2821 aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac
     2881 agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg
     2941 catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg
     3001 gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt
     3061 caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc
     3121 ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc
     3181 cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg
     3241 tcgaaaaggt gtctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg
     3301 gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta
     3361 agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat
     3421 taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga
     3481 gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg
     3541 gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg
     3601 gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc
     3661 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     3721 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     3781 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     3841 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     3901 accgcacaga tgcgtaagga gaaaataccg cataaggcgc tcttccgctt cctcgctcac
     3961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4561 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4741 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatg atatatctcc
     4801 caatttgtgt agggcttatt atgcacgctt aaaaataata aaagcagact tgacctgata
     4861 gtttggctgt gagcaattat gtgcttagtg catctaacgc ttgagttaag ccgcgccgcg
     4921 aagcggcgtc ggcttgaacg aatttctagc tagacattat ttgccgacta ccttggtgat
     4981 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcacgagg ccaagcgatc
     5041 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     5101 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     5161 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     5221 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     5281 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     5341 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     5401 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     5461 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     5521 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     5581 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     5641 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     5701 tacctctgat agttgagtcg atacttcggc gatcaccgct tcccccatga tgtttaactt
     5761 tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg
     5821 acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa
     5881 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     5941 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagcttac gaaccgaaca
     6001 ggcttatgtc cactgggttc gtgcccgaat tgatcacagg cagcaacgct ctgtcatcgt
     6061 tacaatcagc atgctaccct ccgcgagatc atccgtgttt aaaacccggc agcttagttg
     6121 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
     6181 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     6241 gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga
     6301 cgcttagaca acttaataac acattgcgga tccctcggtc acatgtgcat cctcgtaaag
     6361 cgactaaatg gagcaaccta ctgtttttga agtttgggtt gccatgtaat cacttgcaat
     6421 aaatatatgt actaatgact gcatcggctt gcgtgtctaa atttgtagct tattgaaaac
     6481 tttttgtggt taaagacgaa tccagttaac taacataagt ttaacaaaac aagacctttt
     6541 ccatatatag accaccgtta ccaacttaca aagaagccaa ataaagtaca acacaacact
     6601 tgcaagttac aaaaagagag gaaaaaaaac atggtaccat ttgacaacat atagaaacac
     6661 acataaacat tacttataac aaaaagattc aaactcggct tttattatct ccccctgatc
     6721 aaattctaaa cactttattc ctcataaact aactccccga cattggagtt attggactat
     6781 gacgatgaca tcagtggggt aagctcactt gtgccccagt ttgctaggga ggtcgcagta
     6841 tctggccaca gccacctcgt gctgctcgac gtaggtttct ttgtcggcct ccttgattct
     6901 ttccagtctg tggtccacat agtagacgcc gggcatcttg aggttcttag cgggtttctt
     6961 ggatctgtat gtggtcttga agttgcagat caggtggccc ccgcccacga gcttcagggc
     7021 catgtcgctt ctgccttcca ggccgccgtc agcggggtac agcatctcgg tgttggcctc
     7081 ccagccgagt gttttcttct gcatcacagg gccgttggat gggaagttca cccctctgat
     7141 cttgacgttg tagatgaggc agccgtcctg gaggctggtg tcctgggtag cggtcagcac
     7201 gcccccatct tcgtatgtgg tgactctctc ccatgtgaag ccctcaggga aggactgctt
     7261 aaagaagtcg gggatgccct gggtgtggtt gatgaaggtt ctgctgccgt acatgaagct
     7321 ggtagccagg atgtcgaagg cgaaggggag agggccgccc tcgaccacct tgattctcat
     7381 ggtctgggtg ccctcgtagg gcttgccttc gccctcggat gtgcacttga agtggtggtt
     7441 gttcacggtg ccctccatgt acagcttcat gtgcatgttc tccttaatca gctcgctcat
     7501 agtagtgtgc tggccaccac gagtacggtc acggtcatgt tccccaccag tatgttgctg
     7561 tccgtagtac tgagctctgt ctttcagatc ctgagctttg cttcccaact tcatccttgc
     7621 actgtccaac ttgtctgatc cctgtgggtg ctctcccgtt gcgtacctat tcacaatcaa
     7681 taggtatcat accaagcatt ttacctatat agatgatata cacagacaca ttcaagctag
     7741 ctaggtcaaa tttcgtattt agttaaccaa atgttcccta gaatttgcaa tttcttattg
     7801 ttacatttgt tattcgacca aaaaaattga tccaaagcta ctggctcaac acatgcatgc
     7861 acatattgca caaaattatg aagtaagatg ataaatgtgt gcttacttgt aaatccaaga
     7921 gaaaacggtt atagcggcaa tgccaaaccc tccagaggaa agaaaaccgg tgatgaggag
     7981 tgcaactgtg atgagagccg ggacaaggat tgggctgaag ataacgagca gaggtgttgc
     8041 aacagtcaaa gctatgacag ttccaacaag ggtaaggctg gagagaacaa ggagggaacc
     8101 accagctgtg acagcagttg cagctttagc aatctgccta gacttggagt agtcagatcc
     8161 tcgtccggac atctggtact ggtcacgatc tcggcccatc atcgggtact gatctctgcc
     8221 gatgatatcg tgatgggttc ctctagctgt atccgccatt tttttgttct tgtttactag
     8281 agagaatgag attttgaatg agagattgat caaaaagatg tgtatatatt ttggaagagg
     8341 taggagtgtt tgttagatat attaagaaag gagacgagaa gtcacgtgtc aatggagaca
     8401 tcatgcaatt gtaggccacg tgttaatggc gtcaccatgc agccgctgca tgtgttaacg
     8461 agaggagtct attcaaagac acgtttgttt tttgcctagc tcagaatgtg gcgttggcat
     8521 cgaaccgaaa cataaaacgc aaaacgaacc tttgggctcg tcagtatggg tttgggcctt
     8581 caatagattc cttctatacg gcccacatat ccactattgg caacaaaatt agaaaaattt
     8641 aggatttagt tttaaaattt gttccgcgac attctagaac tccgagctcg aattctagta
     8701 gcgcactatt ttcagaagaa gttcccaata gtagtccaaa atttttgtaa cgaagggagc
     8761 ataatagtta catgcaaagg aaaactgcca ttctttagag gggatgcttg tttaagaaca
     8821 aaaaatatat cactttcttt tgttccaagt cattgcgtat ttttttaaaa atatttgttc
     8881 cttcgtatat ttcgagcttc aatcacttta tggttctttg tattctggct ttgctgtaaa
     8941 tcgtagctaa ccttcttcct agcagaaatt attaatactt gggatatttt tttagaatca
     9001 agtaaattac atattaccac cacatcgagc tgcttttaaa ttcatattac agccatatag
     9061 gcttgattca ttttgcaaaa tttccaggat attgacaacg ttaacttaat aatatcttga
     9121 aatattaaag ctattatgat taggggtgca aatggaccga gttggttcgg tttatatcaa
     9181 aatcaaacca aaccaactat atcggtttgg attggttcgg ttttgccggg ttttcagcat
     9241 tttctggttt ttttttttgt tagatgaata ttattttaat cttactttgt caaatttttg
     9301 ataagtaaat atatgtgtta gtaaaaatta atttttttta caaacatatg atctattaaa
     9361 atattcttat aggagaattt tcttaataac acatgatatt tatttatttt agtcgtttga
     9421 ctaatttttc gttgatgtac actttcaaag ttaaccaaat ttagtaatta agtataaaaa
     9481 tcaatatgat acctaaataa tgatatgttc tatttaattt taaattatcg aaatttcact
     9541 tcaaattcga aaaagatata taagaatttt gatagatttt gacatatgaa tatggaagaa
     9601 caaagagatt gacgcatttt agtaacactt gataagaaag tgatcgtaca accaattatt
     9661 taaagttaat aaaaatggag cacttcatat ttaacgaaat attacatgcc agaagagtcg
     9721 caaatatttc tagatatttt ttaaagaaaa ttctataaaa agtcttaaag gcatatatat
     9781 aaaaactata tatttatatt ttggtttggt tcgaatttgt tttactcaat accaaactaa
     9841 attagaccaa atataattgg gatttttaat cgatgctagc ttgtttacac ctcgattgtt
     9901 tcacatatca tttattacct cctattagta catgtgtgaa cgtacggaat aactttgccc
     9961 ccaaaattta aaagaacatg cagttcttta caatgtattg attatatatt atataaggta
    10021 gttggattgg ttttgattta ggtttcgaaa aacagattga gtttaattgt gggataatat
    10081 gtttggaaca attttttgag aaagtaattt taaagaattt ttatgtaact ttcatgtgaa
    10141 aatctactat tgaattgttt tttgttaaaa caatctatat gatttacaaa aagttatcta
    10201 aacactaaag acattgagta ttctttgagt ataaatgaac caagtggagg aatacgatca
    10261 tcgcctttaa aatttaagac tggagctaat atgaacccca attactggag ctaattggat
    10321 atttaattac tgggtccttg aggtgggtta ttaagaggta ttaatttgag taataaattt
    10381 taagatgagc ctttaaaaat caaaaataag aaaaaggttt gggggcttaa gatgaagcca
    10441 catgtaatag gaaagctggg tccaggggtt tgatagatgg aggggtaaat acttttaaat
    10501 aggtaacgat tggggtagaa aatatcgaat agtataacat gaatacttat aactaatatt
    10561 tgatagtaga gggacatatc ccttttgtgt ttcttatttg gataccaaaa aaatcagacc
    10621 aaaatgaact gacaagctct ttatccatat ataccaaatc acaacatcat ttcacataca
    10681 actctatata aaaatattaa ttttaaattg actcttaaca tgaactgctt gatttttcat
    10741 accaacaatg accgaatatt tgagattgac tcaaacaaag ggatagaaac aaaaataatc
    10801 aaaaggaaat acaaatgaga agcatattat ctgcaatcaa tagacgaggt gctgctctca
    10861 aaatgaaagt ttgtatattt cttctgtttc tctttctggg aatgctgtat ggtcactgga
    10921 ttttggtttt aggaattaga aattttattg atagaagtat tttacaaata caaatacata
    10981 ctaagggttt cttatatgct caacacatga gcgaaaccct ataagaaccc taattccctt
    11041 atctgggaac tactcacaca ttattctgga gaaaaataga gagagataga tttgtagaga
    11101 gagactggtg atttttgcgg acaagctcag actttacgct tcttctttgg gtcagcccta
    11161 gagtcaccac ccaactgcga cagatctatc cttgtctcat agagccctgt aatagattgg
    11221 tgaataaggg tagcgtccaa cacttccttg gttgatgtgt agcgctttct gtcgatagtg
    11281 gtatcgaagt acttaaaggc tgctggggct cccagattgg tcagtgtgaa caggtggatg
    11341 atgttttccg cttgttccct tatcggctta tctctgtgct tgttgtacgc ggaaagcacc
    11401 tttaattgta gaaaaccagt aaaacatgga caattaggag gcatcgatta tctaaccaaa
    11461 gactttaaca acactaatct gccactaata ttcatgagaa gtccttacct tatcgaggtt
    11521 agcgtctgca agaatgaccc gcttggaaaa ttcgctgatc tgctctatga tctcatcaag
    11581 atagtgcttg tgctgctcga caaacaactg cttctgctcg ttgtcctctg gtgacccctt
    11641 cagcttctca tagtgtgagg ccaagtacag gaagttgacg tacttactgg gcaatgccag
    11701 ctcgttgccc ttctgcagtt ctccggcaga tgccagcatg cgttttctgc cattttccaa
    11761 ttcgaaaagc gaatacttgg gcagtttgat aatgaggtct ttcttcacct ctttgtaacc
    11821 tggtttgtat aattttggtt cagttaatta ggaaaattac ataatggctc caaaaaaaaa
    11881 ttatcgacct tttaccaaaa aaaataaaat ttatcgacct tgacaggaaa aagttacaaa
    11941 tcaaaagaaa gaatatcgtc tggcatccaa tattttacct ttagcctcga ggaagtcgat
    12001 cggattcttc tcgaacgagg acctttccat tatggtaatt ccaagcagtt ccttgacgct
    12061 tttcagtttc ttgctcttgc ccttctccac cttggccact accagtacgc tgtatgccac
    12121 ggtagggcta tcaaagcctc cgtatttctt cggatcccaa tccttcttcc tcgcaatcag
    12181 tttgtcggag ttgcgcttag ggagaatgga ctccttggag aaaccaccgg tttgaacctc
    12241 cgttttcttg actatgttga cttggggcat gctgagaacc ttacgcacag tcgcaaaatc
    12301 cctgccctta tcccagacaa tctcacctgc aagaaacagc aaagagagag ttaagatata
    12361 agtgaacaag aaagtcttgg gatcgacata catacaatca attgatcaaa ttcctaggat
    12421 taaacatatg aaataggaaa gacagacctg tttcgccatt tgtctcgatc agaggcctct
    12481 tccggatttc accattcgcc agagtgatct ctgtcttgaa gaagttcatg atattcgagt
    12541 aaaagaagta tttcgcggta gccttgccga tctcttgttc tgacttcgcg atcatcttcc
    12601 taacgtcgta caccttgtaa tccccgtaca caaactcaga ctccagtttc ggatacttct
    12661 tgatgagagc agtgccaaca acggcgttca ggtaggcgtc atgggcatga tggtagttgt
    12721 tgatctcgcg aaccttgtaa aactggaagt ccttccggaa gtcactaacc agtttgctct
    12781 tgagggtgat cacctgcgta accatcaaaa gtcaatatgg attcatcaca aattaatcag
    12841 gaatagaaag atcgggtggc cttgaagttg atcaaataaa aagtttcgaa ctttagtttg
    12901 atatggtaaa ggaaagttgg aaactttacc tttacctccc tgataagttt gtcattctca
    12961 tcatacttcg tattcatacg cgaatcgagt atctgggcca catgtttcgt aatctgacga
    13021 gtttcaacca actggcgctt gatgaagccg gctttgtcga gctcagacag tccgcccctt
    13081 tctgccttcg tgagattgtc gaatttccgt tgggttatga gcttagcatt caaaagctgc
    13141 ctccaatagt tcttcatctt cttcacaacc tcctcgctgg gtacattgtc ggattttccg
    13201 cggttcttgt ctgagcgcgt aaggacctat aacacgggag aaaccaaaaa gcaaactcat
    13261 ttcacgccaa atactacaga tttctcaaaa attaagttat aatccatata tcctgacaag
    13321 aaattgatta aaacacagtt gctttacctt gttgtcaata ctatcgtcct tcaggaaaga
    13381 ttgagggaca atgtgatcca catcatagtc ggagagtctg ttaatgtcga gctcttggtc
    13441 gacgtacata tctctcccat tctggaggta atagaggtac agcttctcgt tctggagttg
    13501 agtattttcg actgggtgct ctttcagaat ctgactccca agctccttga tgccttcctc
    13561 tatccgcttc atgcgctccc gtgagttctt ctgccctttc tgagtcgtct ggttttccct
    13621 cgccatctcg atgacgatgt tctccggctt atggcggccc atgacctgta gcaataatta
    13681 aaacaaaaat accacaaaca gttagaactg aggcaagaag aatgaatgca gagaaaaaaa
    13741 atcttagcta ttcatggaaa tggaagtgcg acgaacacct aaaatgcaag gagcataacc
    13801 aaatgcagaa cttaccttca ccagctcatc cacaaccttc actgtctgca gtatgccctt
    13861 ctttatagct ggcgatccag ccaggttggc tatgtgttcg tgaaggctat cgccctgacc
    13921 tgacacttgg gccttctgga tatcttcttt gaaggtcagt gagtcatcgt gaataagctg
    13981 catgaagttc cgattcgcaa aaccgtcgga tttgagaaag tccagtatag tcttgccgga
    14041 ttgcttgtcg cggatgccgt tgatgagctt cctggagagc cttccccaac ctaaaacata
    14101 gaacaaacat ttgtctcatc acaaatatgc atcaaggaac actcagaagt tagtcatata
    14161 cccaaattct tgaaactaga gaaacaaaaa aagagggatc attgcggaaa ccttttgacc
    14221 tcttacctgt atatctcctc ctcttcaact gcttcatgac cttgtcatcg aacagatgcg
    14281 cataagtctt aaggcgctcc tcgatcattt ctctgtcctc aaacagcgtg agagtcagca
    14341 cgatgtcctc gaggatgtcc tcgttctcct cgttgtcaag gaagtccttg tccttgatga
    14401 ttttcaacag gtcatggtat gtgccaagag aggcattgaa cctatcctca acgccgctga
    14461 tttccacaga atcaaagcat tcgatcttct taaagtagtc ctctttaagc tgtttcacag
    14521 tgaccttccg attggttttg aacaagagat cgacaatggc cttcttctgc tcacctgtta
    14581 taacagtaaa aattacttta gaaacaaaac aaaagtgtgg tcttttagga atgattattg
    14641 atcagttcat agtttaaaat cttgtgtttt tatcgtaaaa gtgtggacca ggatttacct
    14701 gacaggaagg ccggtttgcg catcccctcc gtcacatatt tcaccttcgt cagctcgttg
    14761 tagaccgtga agtattcgta caggagacta tgcttgggca ggactttctc atttgggaga
    14821 ttcttatcga agttggtcat cctttcgatg aaggactgtg cactcgcacc cttgtccacg
    14881 acctcttcga agttccaagg cgtgatagtc tcctcggatt tgcgtgtcat ccacgcaaac
    14941 cgcgagttcc cgcgtgcgag tggacctaat gaattaatag aaaaatcagg caaattagaa
    15001 aagaaataat tctcaaaaga gaacaactga ggtcagaaaa tagcattcat caactgaaac
    15061 taggtatttc ttcatatgat actaacctac atagtaggga atgcggaacg tgaggatctt
    15121 ttcaatcttc tctctgttgt ctttcaggaa cggatagaag tcctcttgcc tcctcaagat
    15181 agcatgcagc tcaccgaggt ggatctggtg cggtatgctg ccattgtcga aggtacgctg
    15241 cttccgaagc aaatcttcgc ggttcaattt aacgagcagc tcctctgtac cgtccatctt
    15301 ttccagaata ggcttgatga atttgtagaa ttcctcctga cttgcaccta ataatatcat
    15361 cagaggaaaa ctcaattctt atatcttaca gaaatgaaca acaaaataca tgtagcagag
    15421 aggaaaacaa tcataagtaa caacttacct ccatcaatgt accctgcgta gccattcttc
    15481 gattggtcaa agaaaatctc cttgtacttc tcggggagtt gctggcgaac caaagccttc
    15541 agaagggtca gatcctggtg atgctcgtcg tacctcttga tcatggaggc gctaagtggc
    15601 gctttggtta tctccgtgtt gacccgcaga atgtcactga gcaagatagc gtctgacaga
    15661 ttcttggcag ccaagaacag gtcggcatac tgatcaccta caaaagttta aaagactcaa
    15721 ttttaactct acagcttcaa ataaatttcc gcaaagaaat tgataggttt agacaacaaa
    15781 agaagagttc caatttcaag agcacctatt tgggcgagca gattgtccag atcatcatcg
    15841 tatgtatcct ttgacagctg cagcttcgcg tcctcggcga ggtcaaagtt gcttttgaag
    15901 ttaggggtga ggcccagaga cagtgcaatc agattcccga acaagccgtt cttcttctcg
    15961 cctggcaatt gagcgatcag attctcaagc cgtctggact tcgaaagtct ggcagacagt
    16021 attgccttgg cgtcgacacc tgagattgtt tgcaaagaaa aatctcaaag catgtacaat
    16081 ctataaactt atgatggaaa aagaaaagaa aggtaactaa ggaatgttac ctgatgcatt
    16141 gatggggttc tcctcgaaca gctgattgta ggtttggacg agctgaatga acagtttatc
    16201 cacatcggag ttatccgggt tgaggtcccc ttcgatcaag aaatggccac gaaactttat
    16261 catgtgcgcg agagccaggt atatgagccg aaggtctgcc ttatcggttg agtccaccag
    16321 cttcttgcga aggtggtata tagtcggata cttttcatga taggctactt cgtcaacaat
    16381 attcccaaag atggggtgac gctcgtgttt cttatcttcc tctaccagga aggattcctc
    16441 gaggcgatgg aagaacgagt cgtcaacctg caaaatcaag tttattgatc atcaagtgag
    16501 tttacatgta gttaaatgct ttcagtaata tcccaatgaa gcaggatatg ctttcagtac
    16561 aaaatacata agcatactat atcataaaaa tccttacctt cgccatctcg ttgctaaaga
    16621 tctcttggag ataacagatt ctgttcttcc tgcgagtgta ccttctcctc gcagtacgtt
    16681 taagccttgt cgcctccgcg gtttctccag agtcaaagag cagagctccg atcaagttct
    16741 tcttaatgct gtgcctatct gtgttgccca gcactttaaa tttcttcgaa ggtaccttgt
    16801 attcatcggt gataacggcc cacccaactg agttcgtacc aatatcaagt ccaatgctgt
    16861 acttcttgtc cattttccag ctgaccttcc gcttcttctt cggtgggcta gaagccattg
    16921 ctgtggtgag agaaacagag cgtgagctca aatacacaag tacgaaatca gagttatgag
    16981 agaggatttg gagaaagaag atcacctgaa gatcgtttgc gtcgcgcgaa tgagagagac
    17041 ggcagagaaa acgagaagct ttaagaggac gaacaaaaac cctaatgtca ctccttttat
    17101 atagatgaga aatttagggt ttgggctcta atggagctgt ctacatgggc tttcatttct
    17161 ttatccagta aagaaccggt ttaacaggtt ccctggttta tacgtggttt tagccggtta
    17221 acttacggaa tattattttt atcttaagtt acacaaatta catcattaat agtaaaacaa
    17281 aattttatgg tgcaatttaa gtttttatga tgagtgaaat tcgaataata tacaaatatt
    17341 ttactgttta ggtttcccta aacctacttt tagatatgta aattaaacat tttttttttt
    17401 ttggttagaa ccaagtcagt gatcatagaa ctcaaaaagc tggtgtttca attggtacaa
    17461 gcatttttat tgcaagatat aaaaattcag aaggacatat ccttgatgta tgtacatgaa
    17521 ggaaccgatt ttgactccaa gctacaaaga agaagaagaa tgatatacaa agaaaactgc
    17581 atatcctcac catgaaattg aaagctagag gaaggggaat aacagatact ctgcgtagaa
    17641 gagattccat ataaaccgat aatatcctga gatagcttcc tgcaataatc aaatgtttct
    17701 acactccgtt agatatacac tccattgatg ttccgtgata acgagtttat gtttgtacaa
    17761 aagtatttta ccttggtgta gtttgctttt tctagtaccc atgtctacca aagatcaaca
    17821 acaaagatat tagagattaa tgaatttggc agtgtcaaat gcagaaacta aagcatatag
    17881 ttatgaagtt cttgtgcatt atctcaaaag tatgtaagag atataagagt atctcataat
    17941 gctttacctg gaaaattaag cctgaagcca agatcacatg tgcaggaatc atcaagctac
    18001 ctctaaaaac ctgttcattg ttaaaggata agagggttgg tcttttcaaa gctgcatact
    18061 gaggaaagta acaatatcgc aacaaaaata aattggtggc gatagtttat caagcttcta
    18121 catgcagtaa tggtgttcgt gctttaatta accgaaatag ttaagacgga aatagatgaa
    18181 ttcacctgag gcatgtagaa agctagtgat atggctgaaa cataatttac tagcagaagt
    18241 ccagaaccga ggaatgcaat gtttctcact ccaagttttg ttgccagggt tgatatttgg
    18301 aacctattca agatcaatca aaggcatgat taactaaaca tgcttgaaca aacacataaa
    18361 gacaatatat caaacggagc tggttaaaca ttgtctttga ctgtcaaatg gattgagcca
    18421 aagagcggaa agaaaatggt tatgcgtgca gttacttact ttcgatctcc ttcaacatca
    18481 ggaaggtcct ttgtaatagc aatgaccagt gcaaacagtg tcacaaaaga tgtgatgaac
    18541 gccacaggtg cactgcaaat agcgaatcaa aagttgagct ccaagcttga attcttgcag
    18601 agaccgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
    18661 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
    18721 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
    18781 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
    18841 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
    18901 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
    18961 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
    19021 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
    19081 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
    19141 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
    19201 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
    19261 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
    19321 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggggggc gtaaacgccg
    19381 cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
    19441 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga
    19501 agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga
    19561 tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc
    19621 cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat
    19681 gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct
    19741 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
    19801 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc
    19861 gtgaacttta cccagtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
    19921 tggccagtgt gccggtttcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
    19981 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta
    20041 tacacagcca gtcggtctca gcttctgcag gcatgcgacg tcgagcttag cttgagcttg
    20101 gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag gatatattgg
    20161 cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga
    20221 aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat
    20281 caaagtactt taaagtactt taaagtactt taaagtactt tgatccaacc cctccgctgc
    20341 tatagtgcag tcggcttctg acgttcagtg cagccgtgtt ctgaaaacga catgtcgcac
    20401 aagtcctaag ttacgcgaca ggctgccgcc ctgccctt
//
