LOCUS       sequence-485609-       16862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Base editor targets to ALS Pro197 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16862)
  AUTHORS   Liu MS, Huang TK, Wang YC, Wang SC, Wu CH, Kuo CH, Lai EM
  TITLE     Floral stage optimization and immune evasion enhance 
            Agrobacterium-mediated genome editing in Arabidopsis.
  JOURNAL   New Phytol. 2025 Nov 30. doi: 10.1111/nph.70795.
  PUBMED    41321035
REFERENCE   2  (bases 1 to 16862)
  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: "New Phytol.
            2025 Nov 30. doi: 10.1111/nph.70795."
COMMENT     Sequence Label: EML8359_pA3A-CBECas9-ALSpro197
FEATURES             Location/Qualifiers
     source          1..16862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        699..774
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2992..3057
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3064..3084
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3109..3132,3133..3135,3136..7209)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             7122..7130
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7210..7257
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7279..7527
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7540..7560
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8217..8233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8217..8233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8230..8252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8241..8257
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8265..8271,8272..8289,8290..8295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8310..8331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8522..9198
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9135..9155
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9265..10290
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    10331..10505
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10583..10607
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             11032..11826
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11913..12501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12402..12421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12655..12672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12687..12827
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12753..12772)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12913..12935
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13171..13365
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13431..14504)
                     /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"
     CDS             complement(14933..15562)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
      301 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
      361 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
      421 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
      481 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
      541 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
      601 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
      661 gagtcgaagt agtgattgca agtccctcgt cgtatgatgt tttagagcta gaaatagcaa
      721 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      781 ttgcaaaatt ttccagatcg atttcttctt cctctgttct tcggcgttca atttctgggg
      841 ttttctcttc gttttctgta actgaaacct aaaatttgac ctaaaaaaaa tctcaaataa
      901 tatgattcag tggttttgta cttttcagtt agttgagttt tgcagttccg atgagataaa
      961 ccaataccat ggttatacta gtgaataaaa gcatttgcgt ttggtttatc attgcgttta
     1021 tacaaggaca gagatccact gagctggaat agcttaaaac cattatcaga acaaaataaa
     1081 ccattttttg ttaagaatca gagcatagta aacaacagaa acaacctaag agaggtaact
     1141 tgtccaagaa gatagctaat tatatctatt ttataaaagt tatcatagtt tgtaagtcac
     1201 aaaagatgca aataacagag aaactaggag acttgagaat atacattctt gtatatttgt
     1261 attcgagatt gtgaaaattt gaccataagt ttaaattctt aaaaagatat atctgatcta
     1321 ggtgatggtt atagactgta attttaccac atgtttaatg atggatagtg acacacatga
     1381 cacatcgaca acactatagc atcttattta gattacaaca tgaaattttt ctgtaataca
     1441 tgtctttgta cataatttaa aagtaattcc taagaaatat atttatacaa ggagtttaaa
     1501 gaaaacatag cataaagttc aatgagtagt aaaaaccata tacagtatat agcataaagt
     1561 tcaatgagtt tattacaaaa gcattggttc actttctgta acacgacgtt aaaccttcgt
     1621 ctccaatagg agcgctactg attcaacatg ccaatatata ctaaatacgt ttctacagtc
     1681 aaatgcttta acgtttcatg attaagtgac tatttaccgt caatcctttc ccattcctcc
     1741 cactaatcca actttttaat tactcttaaa tcaccactaa gctagtaacg cctatcatga
     1801 attagctcta ctaaatctag caacctttca aatttgcagt attgcaggtg tctctgtgtc
     1861 tttaaaatag ttgccttatg atttcttcgg tttcaagatg atcaaatagt tatagatttc
     1921 atgctcacac atgctcatta gatgtgtaca tactttactt acccaaatct attttctcgc
     1981 aaagattttg atggtaaagc tgatttggtt ctattgaact aaatcaaacg agtttcagac
     2041 tgagtgattc taatccggcc cattagcccc taaacagacc cactaattac gcagctttta
     2101 atagagtaat tacacctagt ttacccacta aaccactaag cactaattat ctcacaatct
     2161 aatgagcttc cctcgtaatt acttgggctt tcactctacc atttatttgt aacagtcaag
     2221 tctctactgt ctctatataa actctctaaa gttaacacac aattctcatc acaaacaaat
     2281 caaccaaagc aacttctact ctttcttctt tcgaccttat caatctgttg agaaatctag
     2341 atggaggcct ctccggcttc aggtcctcgc catctcatgg acccccatat ctttacctcc
     2401 aatttcaata acggcattgg caggcacaag acatacctct gttatgaggt cgagcggctc
     2461 gataacggaa cctcagtgaa gatggaccaa cacaggggtt tccttcataa ccaagcgaaa
     2521 aatctccttt gcggtttcta tgggcggcac gcggagctga gattcttgga tttggtcccg
     2581 agcctccagc tggatcccgc gcaaatatac cgcgtcactt ggttcatatc atggagcccg
     2641 tgctttagct gggggtgtgc cggtgaggtt cgcgcgttcc tccaggaaaa cacccatgtt
     2701 cgcctccgga ttttcgcagc gaggattttc gactatgatc cactttacaa agaggccctc
     2761 caaatgctta gagatgctgg ggctcaagtt tcaattatga catacgacga gtttaagcat
     2821 tgttgggaca cttttgtcga tcatcagggt tgtcccttcc agccgtggga tggtcttgat
     2881 gagcattcgc aagctttgag tggcaggctt cgcgcgatac ttcagaacca gggtaactca
     2941 ggttctgaga ctcctggcac aagtgagtca gcaacacccg agtccaccat ggattacaag
     3001 gaccacgacg gggattacaa ggaccacgac attgattaca aggatgatga tgacaagatg
     3061 gctccgaaga agaagaggaa ggttggcatc cacggggtgc cagctgctga caagaagtac
     3121 tcgatcggcc tcgctattgg gactaactct gttggctggg ccgtgatcac cgacgagtac
     3181 aaggtgccct caaagaagtt caaggtcctg ggcaacaccg atcggcattc catcaagaag
     3241 aatctcattg gcgctctcct gttcgacagc ggcgagacgg ctgaggctac gcggctcaag
     3301 cgcaccgccc gcaggcggta cacgcgcagg aagaatcgca tctgctacct gcaggagatt
     3361 ttctccaacg agatggcgaa ggttgacgat tctttcttcc acaggctgga ggagtcattc
     3421 ctcgtggagg aggataagaa gcacgagcgg catccaatct tcggcaacat tgtcgacgag
     3481 gttgcctacc acgagaagta ccctacgatc taccatctgc ggaagaagct cgtggactcc
     3541 acagataagg cggacctccg cctgatctac ctcgctctgg cccacatgat taagttcagg
     3601 ggccatttcc tgatcgaggg ggatctcaac ccggacaata gcgatgttga caagctgttc
     3661 atccagctcg tgcagacgta caaccagctc ttcgaggaga accccattaa tgcgtcaggc
     3721 gtcgacgcga aggctatcct gtccgctagg ctctcgaagt ctcggcgcct cgagaacctg
     3781 atcgcccagc tgccgggcga gaagaagaac ggcctgttcg ggaatctcat tgcgctcagc
     3841 ctggggctca cgcccaactt caagtcgaat ttcgatctcg ctgaggacgc caagctgcag
     3901 ctctccaagg acacatacga cgatgacctg gataacctcc tggcccagat cggcgatcag
     3961 tacgcggacc tgttcctcgc tgccaagaat ctgtcggacg ccatcctcct gtctgatatt
     4021 ctcagggtga acaccgagat tacgaaggct ccgctctcag cctccatgat caagcgctac
     4081 gacgagcacc atcaggatct gaccctcctg aaggcgctgg tcaggcagca gctccccgag
     4141 aagtacaagg agatcttctt cgatcagtcg aagaacggct acgctgggta cattgacggc
     4201 ggggcctctc aggaggagtt ctacaagttc atcaagccga ttctggagaa gatggacggc
     4261 acggaggagc tgctggtgaa gctcaatcgc gaggacctcc tgaggaagca gcggacattc
     4321 gataacggca gcatcccaca ccagattcat ctcggggagc tgcacgctat cctgaggagg
     4381 caggaggact tctacccttt cctcaaggat aaccgcgaga agatcgagaa gattctgact
     4441 ttcaggatcc cgtactacgt cggcccactc gctaggggca actcccgctt cgcttggatg
     4501 acccgcaagt cagaggagac gatcacgccg tggaacttcg aggaggtggt cgacaagggc
     4561 gctagcgctc agtcgttcat cgagaggatg acgaatttcg acaagaacct gccaaatgag
     4621 aaggtgctcc ctaagcactc gctcctgtac gagtacttca cagtctacaa cgagctgact
     4681 aaggtgaagt atgtgaccga gggcatgagg aagccggctt tcctgtctgg ggagcagaag
     4741 aaggccatcg tggacctcct gttcaagacc aaccggaagg tcacggttaa gcagctcaag
     4801 gaggactact tcaagaagat tgagtgcttc gattcggtcg agatctctgg cgttgaggac
     4861 cgcttcaacg cctccctggg gacctaccac gatctcctga agatcattaa ggataaggac
     4921 ttcctggaca acgaggagaa tgaggatatc ctcgaggaca ttgtgctgac actcactctg
     4981 ttcgaggacc gggagatgat cgaggagcgc ctgaagactt acgcccatct cttcgatgac
     5041 aaggtcatga agcagctcaa gaggaggagg tacaccggct gggggaggct gagcaggaag
     5101 ctcatcaacg gcattcggga caagcagtcc gggaagacga tcctcgactt cctgaagagc
     5161 gatggcttcg cgaaccgcaa tttcatgcag ctgattcacg atgacagcct cacattcaag
     5221 gaggatatcc agaaggctca ggtgagcggc cagggggact cgctgcacga gcatatcgcg
     5281 aacctcgctg gctcgccagc tatcaagaag gggattctgc agaccgtgaa ggttgtggac
     5341 gagctggtga aggtcatggg caggcacaag cctgagaaca tcgtcattga gatggcccgg
     5401 gagaatcaga ccacgcagaa gggccagaag aactcacgcg agaggatgaa gaggatcgag
     5461 gagggcatta aggagctggg gtcccagatc ctcaaggagc acccggtgga gaacacgcag
     5521 ctgcagaatg agaagctcta cctgtactac ctccagaatg gccgcgatat gtatgtggac
     5581 caggagctgg atattaacag gctcagcgat tacgacgtcg atcatatcgt tccacagtca
     5641 ttcctgaagg atgactccat tgacaacaag gtcctcacca ggtcggacaa gaaccggggc
     5701 aagtctgata atgttccttc agaggaggtc gttaagaaga tgaagaacta ctggcgccag
     5761 ctcctgaatg ccaagctgat cacgcagcgg aagttcgata acctcacaaa ggctgagagg
     5821 ggcgggctct ctgagctgga caaggcgggc ttcatcaaga ggcagctggt cgagacacgg
     5881 cagatcacta agcacgttgc gcagattctc gactcacgga tgaacactaa gtacgatgag
     5941 aatgacaagc tgatccgcga ggtgaaggtc atcaccctga agtcaaagct cgtctccgac
     6001 ttcaggaagg atttccagtt ctacaaggtt cgggagatca acaattacca ccatgcccat
     6061 gacgcgtacc tgaacgcggt ggtcggcaca gctctgatca agaagtaccc aaagctcgag
     6121 agcgagttcg tgtacgggga ctacaaggtt tacgatgtga ggaagatgat cgccaagtcg
     6181 gagcaggaga ttggcaaggc taccgccaag tacttcttct actctaacat tatgaatttc
     6241 ttcaagacag agatcactct ggccaatggc gagatccgga agcgccccct catcgagacg
     6301 aacggcgaga cgggggagat cgtgtgggac aagggcaggg atttcgcgac cgtcaggaag
     6361 gttctctcca tgccacaagt gaatatcgtc aagaagacag aggtccagac tggcgggttc
     6421 tctaaggagt caattctgcc taagcggaac agcgacaagc tcatcgcccg caagaaggac
     6481 tgggatccga agaagtacgg cgggttcgac agccccactg tggcctactc ggtcctggtt
     6541 gtggcgaagg ttgagaaggg caagtccaag aagctcaaga gcgtgaagga gctgctgggg
     6601 atcacgatta tggagcgctc cagcttcgag aagaacccga tcgatttcct ggaggcgaag
     6661 ggctacaagg aggtgaagaa ggacctgatc attaagctcc ccaagtactc actcttcgag
     6721 ctggagaacg gcaggaagcg gatgctggct tccgctggcg agctgcagaa ggggaacgag
     6781 ctggctctgc cgtccaagta tgtgaacttc ctctacctgg cctcccacta cgagaagctc
     6841 aagggcagcc ccgaggacaa cgagcagaag cagctgttcg tcgagcagca caagcattac
     6901 ctcgacgaga tcattgagca gatttccgag ttctccaagc gcgtgatcct ggccgacgcg
     6961 aatctggata aggtcctctc cgcgtacaac aagcaccgcg acaagccaat cagggagcag
     7021 gctgagaata tcattcatct cttcaccctg acgaacctcg gcgcccctgc tgctttcaag
     7081 tacttcgaca caactatcga tcgcaagagg tacacaagca ctaaggaggt cctggacgcg
     7141 accctcatcc accagtcgat taccggcctc tacgagacgc gcatcgacct gtctcagctc
     7201 gggggcgaca agcggccagc ggcgacgaag aaggcggggc aggcgaagaa gaagaaggga
     7261 gacggatcag gcgggtcaac aaacctcagt gacatcatag agaaggaaac tggtaagcaa
     7321 ctggttattc aagagagcat tctgatgctc cctgaggaag tcgaggaagt tataggaaac
     7381 aagcctgaga gcgatatact cgtgcatacg gcgtatgacg agagcacgga tgaaaatgtg
     7441 atgctcttga ccagcgatgc gccagaatat aaaccctggg cactggttat tcaggactct
     7501 aacggggaaa ataaaatcaa gatgctctca ggtggctccc caaagaagaa acgcaaggtt
     7561 tgagctcaga gctttcgttc gtatcatcgg tttcgacaac gttcgtcaag ttcaatgcat
     7621 cagtttcatt gcgcacacac cagaatccta ctgagtttga gtattatggc attgggaaaa
     7681 ctgtttttct tgtaccattt gttgtgcttg taatttactg tgttttttat tcggttttcg
     7741 ctatcgaact gtgaaatgga aatggatgga gaagagttaa tgaatgatat ggtccttttg
     7801 ttcattctca aattaatatt atttgttttt tctcttattt gttgtgtgtt gaatttgaaa
     7861 ttataagaga tatgcaaaca ttttgttttg agtaaaaatg tgtcaaatcg tggcctctaa
     7921 tgaccgaagt taatatgagg agtaaaacac ttgtagttgt accattatgc ttattcacta
     7981 ggcaacaaat atattttcag acctagaaaa gctgcaaatg ttactgaata caagtatgtc
     8041 ctcttgtgtt ttagacattt atgaactttc ctttatgtaa ttttccagaa tccttgtcag
     8101 attctaatca ttgctttata attatagtta tactcatgga tttgtagttg agtatgaaaa
     8161 tattttttaa tgcattttat gacttgccaa ttgattgaca acgaattcgt aatcatgtca
     8221 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     8281 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     8341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     8401 caacgcgcgg ggagaggcgg tttgcgtatt ggctagagca gcttgccaac atggtggagc
     8461 acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac caaagggcta
     8521 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     8581 tctgtcactt catcaaaagg acagtagaaa aggaaggtgg cacctacaaa tgccatcatt
     8641 gcgataaagg aaaggctatc gttcaagatg cctctgccga cagtggtccc aaagatggac
     8701 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     8761 tggattgatg tgaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat
     8821 acagtctcag aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac
     8881 ctcctcggat tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa
     8941 ggtggcacct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct
     9001 gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac
     9061 gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat
     9121 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat
     9181 ttggagagga cacgctgaaa tcaccagtct ctctctacaa atctatctct ctcgagcttt
     9241 cgcagatccg gggggcaatg agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     9301 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     9361 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     9421 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     9481 ccgattccgg aagtgcttga cattggggag tttagcgaga gcctgaccta ttgcatctcc
     9541 cgccgttcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttcta
     9601 caaccggtcg cggaggctat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     9661 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     9721 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     9781 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     9841 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     9901 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     9961 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
    10021 catccggagc ttgcaggatc gccacgactc cgggcgtata tgctccgcat tggtcttgac
    10081 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
    10141 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
    10201 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
    10261 cccagcactc gtccgagggc aaagaaatag agtagatgcc gaccgggatc tgtcgatcga
    10321 caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga attagggttc
    10381 ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg
    10441 taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc
    10501 agatcccccg aattaattcg gcgttaattc agtacattaa aaacgtccgc aatgtgttat
    10561 taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca gccagccaac
    10621 agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc ccatcagtcc
    10681 gggacggcgt cagcgggaga gccgttgtaa ggcggcagac tttgctcatg ttaccgatgc
    10741 tattcggaag aacggcaact aagctgccgg gtttgaaaca cggatgatct cgcggagggt
    10801 agcatgttga ttgtaacgat gacagagcgt tgctgcctgt gatcaccgcg gtttcaaaat
    10861 cggctccgtc gatactatgt tatacgccaa ctttgaaaac aactttgaaa aagctgtttt
    10921 ctggtattta aggttttaga atgcaaggaa cagtgaattg gagttcgtct tgttataatt
    10981 agcttcttgg ggtatcttta aatactgtag aaaagaggaa ggaaataata aatggctaaa
    11041 atgagaatat caccggaatt gaaaaaactg atcgaaaaat accgctgcgt aaaagatacg
    11101 gaaggaatgt ctcctgctaa ggtatataag ctggtgggag aaaatgaaaa cctatattta
    11161 aaaatgacgg acagccggta taaagggacc acctatgatg tggaacggga aaaggacatg
    11221 atgctatggc tggaaggaaa gctgcctgtt ccaaaggtcc tgcactttga acggcatgat
    11281 ggctggagca atctgctcat gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa
    11341 gatgaacaaa gccctgaaaa gattatcgag ctgtatgcgg agtgcatcag gctctttcac
    11401 tccatcgaca tatcggattg tccctatacg aatagcttag acagccgctt agccgaattg
    11461 gattacttac tgaataacga tctggccgat gtggattgcg aaaactggga agaagacact
    11521 ccatttaaag atccgcgcga gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa
    11581 cttgtctttt cccacggcga cctgggagac agcaacatct ttgtgaaaga tggcaaagta
    11641 agtggcttta ttgatcttgg gagaagcggc agggcggaca agtggtatga cattgccttc
    11701 tgcgtccggt cgatcaggga ggatatcggg gaagaacagt atgtcgagct attttttgac
    11761 ttactgggga tcaagcctga ttgggagaaa ataaaatatt atattttact ggatgaattg
    11821 ttttagtacc tagaatgcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
    11881 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
    11941 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    12001 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactatc
    12061 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    12121 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    12181 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    12241 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    12301 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    12361 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    12421 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    12481 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    12541 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    12601 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    12661 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
    12721 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    12781 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
    12841 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
    12901 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
    12961 cgaggcaggg tgccttgatg tgggcgccgg cggtcgagtg gcgacggcgc ggcttgtccg
    13021 cgccctggta gattgcctgg ccgtaggcca gccatttttg agcggccagc ggccgcgata
    13081 ggccgacgcg aagcggcggg gcgtagggag cgcagcgacc gaagggtagg cgctttttgc
    13141 agctcttcgg ctgtgcgctg gccagacagt tatgcacagg ccaggcgggt tttaagagtt
    13201 ttaataagtt ttaaagagtt ttaggcggaa aaatcgcctt ttttctcttt tatatcagtc
    13261 acttacatgt gtgaccggtt cccaatgtac ggctttgggt tcccaatgta cgggttccgg
    13321 ttcccaatgt acggctttgg gttcccaatg tacgtgctat ccacaggaaa cagacctttt
    13381 cgaccttttt cccctgctag ggcaatttgc cctagcatct gctccgtaca ttaggaaccg
    13441 gcggatgctt cgccctcgat caggttgcgg tagcgcatga ctaggatcgg gccagcctgc
    13501 cccgcctcct ccttcaaatc gtactccggc aggtcatttg acccgatcag cttgcgcacg
    13561 gtgaaacaga acttcttgaa ctctccggcg ctgccactgc gttcgtagat cgtcttgaac
    13621 aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca ggcggtagag aaaacggccg
    13681 atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca gcacgtccgg gttcttgcct
    13741 tctgtgatct cgcggtacat ccaatcagct agctcgatct cgatgtactc cggccgcccg
    13801 gtttcgctct ttacgatctt gtagcggcta atcaaggctt caccctcgga taccgtcacc
    13861 aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa cgtgcgtggt gtttaaccga
    13921 atgcaggttt ctaccaggtc gtctttctgc tttccgccat cggctcgccg gcagaacttg
    13981 agtacgtccg caacgtgtgg acggaacacg cggccgggct tgtctccctt cccttcccgg
    14041 tatcggttca tggattcggt tagatgggaa accgccatca gtaccaggtc gtaatcccac
    14101 acactggcca tgccggccgg ccctgcggaa acctctacgt gcccgtctgg aagctcgtag
    14161 cggatcacct cgccagctcg tcggtcacgc ttcgacagac ggaaaacggc cacgtccatg
    14221 atgctgcgac tatcgcgggt gcccacgtca tagagcatcg gaacgaaaaa atctggttgc
    14281 tcgtcgccct tgggcggctt cctaatcgac ggcgcaccgg ctgccggcgg ttgccgggat
    14341 tctttgcgga ttcgatcagc ggccgcttgc cacgattcac cggggcgtgc ttctgcctcg
    14401 atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct ccaccaggtc atcacccagc
    14461 gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc tcacgccgat tcctcgggct
    14521 tgggggttcc agtgccattg cagggccggc aggcaaccca gccgcttacg cctggccaac
    14581 cgcccgttcc tccacacatg gggcattcca cggcgtcggt gcctggttgt tcttgatttt
    14641 ccatgccgcc tcctttagcc gctaaaattc atctactcat ttattcattt gctcatttac
    14701 tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc
    14761 gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac ccgcttcatg
    14821 gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg
    14881 ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta actcaaatga
    14941 gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt
    15001 ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga
    15061 tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc tcaccgccga
    15121 tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt ccatccgtga
    15181 cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg tcgtaagggc
    15241 ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca gccaagtccg
    15301 ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc ttcacgtcgc
    15361 ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc cgcacggccg
    15421 cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg gccccggcga
    15481 gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg cctttctggt
    15541 taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta ctcatcgcat
    15601 catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc acctttttag
    15661 acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca gcttggcatc
    15721 agacaaaccg gccaggattt catgcagccg cacggttgag acgtgcgcgg gcggctcgaa
    15781 cacgtacccg gccgcgatca tctccgcctc gatctcttcg gtaatgaaaa acggttcgtc
    15841 ctggccgtcc tggtgcggtt tcatgcttgt tcctcttggc gttcattctc ggcggccgcc
    15901 agggcgtcgg cctcggtcaa tgcgtcctca cggaaggcac cgcgccgcct ggcctcggtg
    15961 ggcgtcactt cctcgctgcg ctcaagtgcg cggtacaggg tcgagcgatg cacgccaagc
    16021 agtgcagccg cctctttcac ggtgcggcct tcctggtcga tcagctcgcg ggcgtgcgcg
    16081 atctgtgccg gggtgagggt agggcggggg ccaaacttca cgcctcgggc cttggcggcc
    16141 tcgcgcccgc tccgggtgcg gtcgatgatt agggaacgct cgaactcggc aatgccggcg
    16201 aacacggtca acaccatgcg gccggccggc gtggtggtgt cggcccacgg ctctgccagg
    16261 ctacgcaggc ccgcgccggc ctcctggatg cgctcggcaa tgtccagtag gtcgcgggtg
    16321 ctgcgggcca ggcggtctag cctggtcact gtcacaacgt cgccagggcg taggtggtca
    16381 agcatcctgg ccagctccgg gcggtcgcgc ctggtgccgg tgatcttctc ggaaaacagc
    16441 ttggtgcagc cggccgcgtg cagttcggcc cgttggttgg tcaagtcctg gtcgtcggtg
    16501 ctgacgcggg catagcccag caggccagcg gcggcgctct tgttcatggc gtaatgtctc
    16561 cggttctagt cgcaagtatt ctactttatg cgactaaaac acgcgacaag aaaacgccag
    16621 gaaaagggca gggcggcagc ctgtcgcgta acttaggact tgtgcgacat gtcgttttca
    16681 gaagacggct gcactgaacg tcagaagccg actgcactat agcagcggag gggttggatc
    16741 aaagtacttt gatcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg
    16801 gataaacctt ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt
    16861 ag
//
