LOCUS       sequence-403581-       19323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-KO with TRE2 exonuclease co-expression in Arabidopsis 
            thaliana.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19323)
  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 19323)
  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: pDGE1177
FEATURES             Location/Qualifiers
     source          1..19323
                     /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(9242..9262)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(14995..15015)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(15031..15087)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     promoter        17498..17600
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             17601..18284
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     primer_bind     complement(17667..17686)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             18604..18909
                     /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     18788..18807
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    19030..19054
                     /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 gcggtacact gccagatgaa gccgaaggag tactttaata tactaggtct ggcgatatga
     8761 tcaatttgaa aatatagagt cataactgta gaaatggaat cagattccat tacaaaaaga
     8821 gataaatctc aactttaaat tttttcagga agaaaaaaat gtttcttcta atcttcgtgt
     8881 tttactggcc ttctcataaa atggtttgat ataaaagaaa attaaataag aaaaagagaa
     8941 aacaaaaggc tagccagaga gaggagaaaa tatataataa tataacttag tttattcatc
     9001 tcataatctc tgcattttgc aaggagatcg aaactgagca aaacatacac aaacagagaa
     9061 ataaaatata gggtacaaac aatatcaata gcttagacaa catgaagcta caaaagaact
     9121 tcgcctcctg tggggcttat tatttactct tttttttttc ttattcttcg tcttttctca
     9181 ttgcctctcg aaattggaaa aagcctttct tttagctcaa cattctgctt ttccaagctc
     9241 agactttacg cttcttcttt gggtcagccc tagagtcacc acccaactgc gacagatcta
     9301 tccttgtctc atagagccct gtaatagatt ggtgaataag ggtagcgtcc aacacttcct
     9361 tggttgatgt gtagcgcttt ctgtcgatag tggtatcgaa gtacttaaag gctgctgggg
     9421 ctcccagatt ggtcagtgtg aacaggtgga tgatgttttc cgcttgttcc cttatcggct
     9481 tatctctgtg cttgttgtac gcggaaagca cctttaattg tagaaaacca gtaaaacatg
     9541 gacaattagg aggcatcgat tatctaacca aagactttaa caacactaat ctgccactaa
     9601 tattcatgag aagtccttac cttatcgagg ttagcgtctg caagaatgac ccgcttggaa
     9661 aattcgctga tctgctctat gatctcatca agatagtgct tgtgctgctc gacaaacaac
     9721 tgcttctgct cgttgtcctc tggtgacccc ttcagcttct catagtgtga ggccaagtac
     9781 aggaagttga cgtacttact gggcaatgcc agctcgttgc ccttctgcag ttctccggca
     9841 gatgccagca tgcgttttct gccattttcc aattcgaaaa gcgaatactt gggcagtttg
     9901 ataatgaggt ctttcttcac ctctttgtaa cctggtttgt ataattttgg ttcagttaat
     9961 taggaaaatt acataatggc tccaaaaaaa aattatcgac cttttaccaa aaaaaataaa
    10021 atttatcgac cttgacagga aaaagttaca aatcaaaaga aagaatatcg tctggcatcc
    10081 aatattttac ctttagcctc gaggaagtcg atcggattct tctcgaacga ggacctttcc
    10141 attatggtaa ttccaagcag ttccttgacg cttttcagtt tcttgctctt gcccttctcc
    10201 accttggcca ctaccagtac gctgtatgcc acggtagggc tatcaaagcc tccgtatttc
    10261 ttcggatccc aatccttctt cctcgcaatc agtttgtcgg agttgcgctt agggagaatg
    10321 gactccttgg agaaaccacc ggtttgaacc tccgttttct tgactatgtt gacttggggc
    10381 atgctgagaa ccttacgcac agtcgcaaaa tccctgccct tatcccagac aatctcacct
    10441 gcaagaaaca gcaaagagag agttaagata taagtgaaca agaaagtctt gggatcgaca
    10501 tacatacaat caattgatca aattcctagg attaaacata tgaaatagga aagacagacc
    10561 tgtttcgcca tttgtctcga tcagaggcct cttccggatt tcaccattcg ccagagtgat
    10621 ctctgtcttg aagaagttca tgatattcga gtaaaagaag tatttcgcgg tagccttgcc
    10681 gatctcttgt tctgacttcg cgatcatctt cctaacgtcg tacaccttgt aatccccgta
    10741 cacaaactca gactccagtt tcggatactt cttgatgaga gcagtgccaa caacggcgtt
    10801 caggtaggcg tcatgggcat gatggtagtt gttgatctcg cgaaccttgt aaaactggaa
    10861 gtccttccgg aagtcactaa ccagtttgct cttgagggtg atcacctgcg taaccatcaa
    10921 aagtcaatat ggattcatca caaattaatc aggaatagaa agatcgggtg gccttgaagt
    10981 tgatcaaata aaaagtttcg aactttagtt tgatatggta aaggaaagtt ggaaacttta
    11041 cctttacctc cctgataagt ttgtcattct catcatactt cgtattcata cgcgaatcga
    11101 gtatctgggc cacatgtttc gtaatctgac gagtttcaac caactggcgc ttgatgaagc
    11161 cggctttgtc gagctcagac agtccgcccc tttctgcctt cgtgagattg tcgaatttcc
    11221 gttgggttat gagcttagca ttcaaaagct gcctccaata gttcttcatc ttcttcacaa
    11281 cctcctcgct gggtacattg tcggattttc cgcggttctt gtctgagcgc gtaaggacct
    11341 ataacacggg agaaaccaaa aagcaaactc atttcacgcc aaatactaca gatttctcaa
    11401 aaattaagtt ataatccata tatcctgaca agaaattgat taaaacacag ttgctttacc
    11461 ttgttgtcaa tactatcgtc cttcaggaaa gattgaggga caatgtgatc cacatcatag
    11521 tcggagagtc tgttaatgtc gagctcttgg tcgacgtaca tatctctccc attctggagg
    11581 taatagaggt acagcttctc gttctggagt tgagtatttt cgactgggtg ctctttcaga
    11641 atctgactcc caagctcctt gatgccttcc tctatccgct tcatgcgctc ccgtgagttc
    11701 ttctgccctt tctgagtcgt ctggttttcc ctcgccatct cgatgacgat gttctccggc
    11761 ttatggcggc ccatgacctg tagcaataat taaaacaaaa ataccacaaa cagttagaac
    11821 tgaggcaaga agaatgaatg cagagaaaaa aaatcttagc tattcatgga aatggaagtg
    11881 cgacgaacac ctaaaatgca aggagcataa ccaaatgcag aacttacctt caccagctca
    11941 tccacaacct tcactgtctg cagtatgccc ttctttatag ctggcgatcc agccaggttg
    12001 gctatgtgtt cgtgaaggct atcgccctga cctgacactt gggccttctg gatatcttct
    12061 ttgaaggtca gtgagtcatc gtgaataagc tgcatgaagt tccgattcgc aaaaccgtcg
    12121 gatttgagaa agtccagtat agtcttgccg gattgcttgt cgcggatgcc gttgatgagc
    12181 ttcctggaga gccttcccca acctaaaaca tagaacaaac atttgtctca tcacaaatat
    12241 gcatcaagga acactcagaa gttagtcata tacccaaatt cttgaaacta gagaaacaaa
    12301 aaaagaggga tcattgcgga aaccttttga cctcttacct gtatatctcc tcctcttcaa
    12361 ctgcttcatg accttgtcat cgaacagatg cgcataagtc ttaaggcgct cctcgatcat
    12421 ttctctgtcc tcaaacagcg tgagagtcag cacgatgtcc tcgaggatgt cctcgttctc
    12481 ctcgttgtca aggaagtcct tgtccttgat gattttcaac aggtcatggt atgtgccaag
    12541 agaggcattg aacctatcct caacgccgct gatttccaca gaatcaaagc attcgatctt
    12601 cttaaagtag tcctctttaa gctgtttcac agtgaccttc cgattggttt tgaacaagag
    12661 atcgacaatg gccttcttct gctcacctgt tataacagta aaaattactt tagaaacaaa
    12721 acaaaagtgt ggtcttttag gaatgattat tgatcagttc atagtttaaa atcttgtgtt
    12781 tttatcgtaa aagtgtggac caggatttac ctgacaggaa ggccggtttg cgcatcccct
    12841 ccgtcacata tttcaccttc gtcagctcgt tgtagaccgt gaagtattcg tacaggagac
    12901 tatgcttggg caggactttc tcatttggga gattcttatc gaagttggtc atcctttcga
    12961 tgaaggactg tgcactcgca cccttgtcca cgacctcttc gaagttccaa ggcgtgatag
    13021 tctcctcgga tttgcgtgtc atccacgcaa accgcgagtt cccgcgtgcg agtggaccta
    13081 atgaattaat agaaaaatca ggcaaattag aaaagaaata attctcaaaa gagaacaact
    13141 gaggtcagaa aatagcattc atcaactgaa actaggtatt tcttcatatg atactaacct
    13201 acatagtagg gaatgcggaa cgtgaggatc ttttcaatct tctctctgtt gtctttcagg
    13261 aacggataga agtcctcttg cctcctcaag atagcatgca gctcaccgag gtggatctgg
    13321 tgcggtatgc tgccattgtc gaaggtacgc tgcttccgaa gcaaatcttc gcggttcaat
    13381 ttaacgagca gctcctctgt accgtccatc ttttccagaa taggcttgat gaatttgtag
    13441 aattcctcct gacttgcacc taataatatc atcagaggaa aactcaattc ttatatctta
    13501 cagaaatgaa caacaaaata catgtagcag agaggaaaac aatcataagt aacaacttac
    13561 ctccatcaat gtaccctgcg tagccattct tcgattggtc aaagaaaatc tccttgtact
    13621 tctcggggag ttgctggcga accaaagcct tcagaagggt cagatcctgg tgatgctcgt
    13681 cgtacctctt gatcatggag gcgctaagtg gcgctttggt tatctccgtg ttgacccgca
    13741 gaatgtcact gagcaagata gcgtctgaca gattcttggc agccaagaac aggtcggcat
    13801 actgatcacc tacaaaagtt taaaagactc aattttaact ctacagcttc aaataaattt
    13861 ccgcaaagaa attgataggt ttagacaaca aaagaagagt tccaatttca agagcaccta
    13921 tttgggcgag cagattgtcc agatcatcat cgtatgtatc ctttgacagc tgcagcttcg
    13981 cgtcctcggc gaggtcaaag ttgcttttga agttaggggt gaggcccaga gacagtgcaa
    14041 tcagattccc gaacaagccg ttcttcttct cgcctggcaa ttgagcgatc agattctcaa
    14101 gccgtctgga cttcgaaagt ctggcagaca gtattgcctt ggcgtcgaca cctgagattg
    14161 tttgcaaaga aaaatctcaa agcatgtaca atctataaac ttatgatgga aaaagaaaag
    14221 aaaggtaact aaggaatgtt acctgatgca ttgatggggt tctcctcgaa cagctgattg
    14281 taggtttgga cgagctgaat gaacagttta tccacatcgg agttatccgg gttgaggtcc
    14341 ccttcgatca agaaatggcc acgaaacttt atcatgtgcg cgagagccag gtatatgagc
    14401 cgaaggtctg ccttatcggt tgagtccacc agcttcttgc gaaggtggta tatagtcgga
    14461 tacttttcat gataggctac ttcgtcaaca atattcccaa agatggggtg acgctcgtgt
    14521 ttcttatctt cctctaccag gaaggattcc tcgaggcgat ggaagaacga gtcgtcaacc
    14581 tgcaaaatca agtttattga tcatcaagtg agtttacatg tagttaaatg ctttcagtaa
    14641 tatcccaatg aagcaggata tgctttcagt acaaaataca taagcatact atatcataaa
    14701 aatccttacc ttcgccatct cgttgctaaa gatctcttgg agataacaga ttctgttctt
    14761 cctgcgagtg taccttctcc tcgcagtacg tttaagcctt gtcgcctccg cggtttctcc
    14821 agagtcaaag agcagagctc cgatcaagtt cttcttaatg ctgtgcctat ctgtgttgcc
    14881 cagcacttta aatttcttcg aaggtacctt gtattcatcg gtgataacgg cccacccaac
    14941 tgagttcgta ccaatatcaa gtccaatgct gtacttcttg tccattttcc agctgacctt
    15001 ccgcttcttc ttcggtgggc tagaagccat tggtccagga ttttcttcaa catctccagc
    15061 ttgcttaaga agagaaaaat tagtagctcc agatccggct tcgaggcttg gaccatcagg
    15121 tggcacgtac atgggctcaa tatgagccca gctgcgggcc tgctcatctg cccaggcgag
    15181 cagctcagga gcacgatgca ggaagatcag aagcagggtg tgcacatcac cttctgctga
    15241 atgggcagca ctgggttcag cctggaagta gcggtggaag agactggcca ggctgtagct
    15301 tttgcggcct tgagccctgg tgccgtggct gtgagcacgg tccaggcccc gcaatgcagg
    15361 cagtgtgtcc aggcagacag tgtcttgggg cagatgggca cccagacgtt gtagctccgt
    15421 gcacagcagt gggaagtcat aatcgaagcc attgtgggcc acaaggcaga tggggccctc
    15481 ctggcggctt aggaagccct gcagtgtcct taccacagcg ccattgaaac cagccttccc
    15541 gcagtgcatc aggctttcgc tgctcaaacc agtaatctca ctggccttgg cagtaaaggg
    15601 gcgctccggg cacatgcaca gtgtgagctt gtccagaaca cggggcagca ccaaggaacc
    15661 agaatcatcc cgttctgggt tctccaggga agagcggtga acagcaaaaa gggatatctc
    15721 tgcaatctca gggtccatgt ttgggagccc agtggcttct aggtccagga atacaaaggt
    15781 ttcagcccga ggtggctcag acatggtgtg gtgagagaaa cagagcgtga gctcaaatac
    15841 acaagtacga aatcagagtt atgagagagg atttggagaa agaagatcac ctgaagatcg
    15901 tttgcgtcgc gcgaatgaga gagacggcag agaaaacgag aagctttaag aggacgaaca
    15961 aaaaccctaa tgtcactcct tttatataga tgagaaattt agggtttggg ctctaatgga
    16021 gctgtctaca tgggctttca tttctttatc cagtaaagaa ccggtttaac aggttccctg
    16081 gtttatacgt ggttttagcc ggttaactta cggaatatta tttttatctt aagttacaca
    16141 aattacatca ttaatagtaa aacaaaattt tatggtgcaa tttaagtttt tatgatgagt
    16201 gaaattcgaa taatatacaa atattttact gtttaggttt ccctaaacct acttttagat
    16261 atgtaaatta aacatttttt tttttttggt tagaaccaag tcagtgatca tagaactcaa
    16321 aaagctggtg tttcaattgg tacaagcatt tttattgcaa gatataaaaa ttcagaagga
    16381 catatccttg atgtatgtac atgaaggaac cgattttgac tccaagctac aaagaagaag
    16441 aagaatgata tacaaagaaa actgcatatc ctcaccatga aattgaaagc tagaggaagg
    16501 ggaataacag atactctgcg tagaagagat tccatataaa ccgataatat cctgagatag
    16561 cttcctgcaa taatcaaatg tttctacact ccgttagata tacactccat tgatgttccg
    16621 tgataacgag tttatgtttg tacaaaagta ttttaccttg gtgtagtttg ctttttctag
    16681 tacccatgtc taccaaagat caacaacaaa gatattagag attaatgaat ttggcagtgt
    16741 caaatgcaga aactaaagca tatagttatg aagttcttgt gcattatctc aaaagtatgt
    16801 aagagatata agagtatctc ataatgcttt acctggaaaa ttaagcctga agccaagatc
    16861 acatgtgcag gaatcatcaa gctacctcta aaaacctgtt cattgttaaa ggataagagg
    16921 gttggtcttt tcaaagctgc atactgagga aagtaacaat atcgcaacaa aaataaattg
    16981 gtggcgatag tttatcaagc ttctacatgc agtaatggtg ttcgtgcttt aattaaccga
    17041 aatagttaag acggaaatag atgaattcac ctgaggcatg tagaaagcta gtgatatggc
    17101 tgaaacataa tttactagca gaagtccaga accgaggaat gcaatgtttc tcactccaag
    17161 ttttgttgcc agggttgata tttggaacct attcaagatc aatcaaaggc atgattaact
    17221 aaacatgctt gaacaaacac ataaagacaa tatatcaaac ggagctggtt aaacattgtc
    17281 tttgactgtc aaatggattg agccaaagag cggaaagaaa atggttatgc gtgcagttac
    17341 ttactttcga tctccttcaa catcaggaag gtcctttgta atagcaatga ccagtgcaaa
    17401 cagtgtcaca aaagatgtga tgaacgccac aggtgcactg caaatagcga atcaaaagtt
    17461 gagctccaag cttgaattct tgcagagacc gagacgttga tcggcacgta agaggttcca
    17521 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
    17581 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
    17641 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
    17701 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
    17761 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
    17821 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
    17881 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
    17941 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
    18001 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
    18061 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
    18121 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
    18181 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
    18241 agtggcaggg ggggcgtaaa cgccgcgtgg atccggctta ctaaaagcca gataacagta
    18301 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
    18361 tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc
    18421 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
    18481 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
    18541 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt
    18601 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
    18661 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
    18721 cgtctgctgt cagataaagt ctcccgtgaa ctttacccag tggtgcatat cggggatgaa
    18781 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tttccgttat cggggaagaa
    18841 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
    18901 ggaatataaa tgtcaggctc ccttatacac agccagtcgg tctcagcttc tgcaggcatg
    18961 cgacgtcgag cttagcttga gcttggatca gattgtcgtt tcccgccttc agtttaaact
    19021 atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
    19081 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
    19141 ccaaccacag ggttcccctc gggatcaaag tactttaaag tactttaaag tactttaaag
    19201 tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc
    19261 gtgttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc
    19321 ctt
//
