LOCUS       sequence-368287-       18408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For plant prime editing in rice plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18408)
  AUTHORS   Xu W, Yang Y, Yang B, Krueger CJ, Xiao Q, Zhao S, Zhang L, Kang G, 
            Wang F, Yi H, Ren W, Li L, He X, Zhang C, Zhang B, Zhao J, Yang J
  TITLE     A design optimized prime editor with expanded scope and capability 
            in plants.
  JOURNAL   Nat Plants. 2022 Jan;8(1):45-52. doi: 10.1038/s41477-021-01043-4. 
            Epub 2021 Dec 23.
  PUBMED    34949802
REFERENCE   2  (bases 1 to 18408)
  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"; doi: 
            "10.1038/s41477-021-01043-4"; journalName: "Nat Plants"; date: 
            "2022-01"; volume: "8"; issue: "1"; pages: "45-52"
COMMENT     Sequence Label: PE-P3-rice
FEATURES             Location/Qualifiers
     source          1..18408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1539..1563
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             3622..3687
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3694..3714
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3724..3744
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3754..3774
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3784..3804
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5956..8469,8470..8472,8473..10056)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             7142..7150
                     /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             10054..10068
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             10075..10095
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10105..10125
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10135..10155
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10165..10185
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10207..10263
                     /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"
     CDS             10264..11289
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     terminator      11300..11552
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    12172..12205
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(12249..12266)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12249..12265)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12258..12280)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    12468..12492
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             13793..14422
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             14851..15924
                     /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      15990..16184
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(16420..16442)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    16528..16668
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     16583..16602
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(16683..16700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16854..17442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16934..16953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(17529..18323)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 atgtttgtca tctggagcca tagaacaggg ttcatcatga gtcatcaact taccttcgcc
       61 gacagtgaat tcagcagtaa gcgccgtcag accagaaaag agattttctt gtcccgcatg
      121 gagcagattc tgccatggca aaacatggtg gaagtcatcg agccgtttta ccccaaggct
      181 ggtaatggcc ggcgacctta tccgctggaa accatgctac gcattcactg catgcagcat
      241 tggtacaacc tgagcgatgg cgcgatggaa gatgctctgt acgaaatcgc ctccatgcgt
      301 ctgtttgccc ggttatccct ggatagcgcc ttgccggacc gcaccaccat catgaatttc
      361 cgccacctgc tggagcagca tcaactggcc cgccaattgt tcaagaccat caatcgctgg
      421 ctggccgaag caggcgtcat gatgactcaa ggcaccttgg tcgatgccac catcattgag
      481 gcacccagct cgaccaagaa caaagagcag caacgcgatc cggagatgca tcagaccaag
      541 aaaggcaatc agtggcactt tggcatgaag gcccacattg gtgtcgatgc caagagtggc
      601 ctgacccaca gcctggtcac caccgcggcc aacgagcatg acctcaatca gctgggtaat
      661 ctgctgcatg gagaggagca atttgtctca gccgatgccg gctaccaagg ggcgccacag
      721 cgcgaggagc tggccgaggt ggatgtggac tggctgatcg ccgagcgccc cggcaaggta
      781 agaaccttga aacagcatcc acgcaagaac aaaacggcca tcaacatcga atacatgaaa
      841 gccagcatcc gggccagggt ggagcaccca tttcgcatca tcaagcgaca gttcggcttc
      901 gtgaaagcca gatacaaggg gttgctgaaa aacgataacc aactggcgat gttattcacg
      961 ctggccaacc tgtttcgggc ggaccaaatg atacgtcagt gggagagatc tcactaaaaa
     1021 ctggggataa cgccttaaat ggcgaagaaa cggtctaaat aggctgattc aaggcattta
     1081 cgggagaaaa aatcggctca aacatgaaga aatgaaatga ctgagtcagc cgagaagaat
     1141 ttccccgctt attcgcacct tccctaatta taacaagacg aactccaatt cactgttcct
     1201 tgcattctaa aaccttaaat accagaaaac agctttttca aagttgtttt caaagttggc
     1261 gtataacata gtatcgacgg agccgatttt gaaaccgcgg tgatcacagg cagcaacgct
     1321 ctgtcatcgt tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc
     1381 agcttagttg ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca
     1441 acggctctcc cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt
     1501 gtgccgagct gccggtcggg gagctgttgg ctggctggtg gcaggatata ttgtggtgta
     1561 aacaaattga cgcttagaca acttaataac acattgcgga cgtttttaat gtaggtacca
     1621 cctaaatttc caagcttgtc gtgcccctct ctagagataa tgagcattgc atgtctaagt
     1681 tataaaaaat taccacatat tttttttgtc acacttgttt gaagtgcagt ttatctatct
     1741 ttatacatat atttaaactt tactctacga ataatataat ctatagtact acaataatat
     1801 cagtgtttta gagaatcata taaatgaaca gttagacatg gtctaaagga caattgagta
     1861 ttttgacaac aggactctac agttttatct ttttagtgtg catgtgttct cctttttttt
     1921 tgcaaatagc ttcacctata taatacttca tccattttat tagtacatcc atttagggtt
     1981 tagggttaat ggtttttata gactaatttt tttagtacat ctattttatt ctattttagc
     2041 ctctaaatta agaaaactaa aactctattt tagttttttt atttaataat ttagatataa
     2101 aatagaataa aataaagtga ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac
     2161 taaggaaaca tttttcttgt ttcgagtaga taatgccagc ctgttaaacg ccgtcgacga
     2221 gtctaacgga caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg aagcagacgg
     2281 cacggcatct ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca ccgttggact
     2341 tgctccgctg tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag ccggcacggc
     2401 aggcggcctc ctcctcctct cacggcaccg gcagctacgg gggattcctt tcccaccgct
     2461 ccttcgcttt cccttcctcg cccgccgtaa taaatagaca ccccctccac accctctttc
     2521 cccaacctcg tgttgttcgg agcgcacaca cacacaacca gatctccccc aaatccmccc
     2581 gtcggcacct ccgcttcaag gtacgccgct cgtcctcccc cccccccctc tctaccttct
     2641 ctagatcggc gttccggtcc atggttaggg cccggtagtt ctacttctgt tcatgtttgt
     2701 gttagatccg tgtttgtgtt agatccgtgc tgctagcgtt cgtacacgga tgcgacctgt
     2761 acgtcagaca cgttctgatt gctaacttgc cagtgtttct ctttggggaa tcctgggatg
     2821 gctctagccg ttccgcagac gggatcgatt tcatgatttt ttttgtttcg ttgcataggg
     2881 tttggtttgc ccttttcctt tatttcaata tatgccgtgc acttgtttgt cgggtcatct
     2941 tttcatgctt ttttttgtct tggttgtgat gatgtggtct ggttgggcgg tcgttctaga
     3001 tcggagtaga attctgtttc aaactacctg gtggatttat taattttgga tctgtatgtg
     3061 tgtgccatac atattcatag ttacgaattg aagatgatgg atggaaatat cgatctagga
     3121 taggtataca tgttgatgcg ggttttactg atgcatatac agagatgctt tttgttcgct
     3181 tggttgtgat gatgtggtgt ggttgggcgg tcgttcattc gttctagatc ggagtagaat
     3241 actgtttcaa actacctggt gtatttatta attttggaac tgtatgtgtg tgtcatacat
     3301 cttcatagtt acgagtttaa gatggatgga aatatcgatc taggataggt atacatgttg
     3361 atgtgggttt tactgatgca tatacatgat ggcatatgca gcatctattc atatgctcta
     3421 accttgagta cctatctatt ataataaaca agtatgtttt ataattattt tgatcttgat
     3481 atacttggat gatggcatat gcagcagcta tatgtggatt tttttagccc tgccttcata
     3541 cgctatttat ttgcttggta ctgtttcttt tgtcgatgct caccctgttg tttggtgtta
     3601 cttctgcagt acgtaagcat ggactacaag gaccacgacg gggattacaa agaccacgac
     3661 atagactaca aggatgacga tgacaaaatg gcaccgaaga aaaaaaggaa ggtcggcggc
     3721 tccccgaaga aaaaaaggaa ggtcggcggc tccccgaaga aaaaaaggaa ggtcggcggc
     3781 tccccgaaga aaaaaaggaa ggtcggaatc catggcgttc cagaaaccct caatatcgag
     3841 gacgagtaca ggctgcatga gacatccaag gagccggacg tgtcactcgg ctctacatgg
     3901 ctgagcgatt tcccacaggc ctgggcggag acaggcggca tgggcctcgc ggtcaggcag
     3961 gcgccgctca tcattccact gaaggcgacc tccacaccgg tcagcatcaa gcagtaccca
     4021 atgtcacagg aggcacggct cggcatcaag ccacacattc agaggctcct ggaccagggc
     4081 attctggtcc cttgccagag cccgtggaac acccctctcc tgccggtgaa gaagcctggc
     4141 acaaatgact acaggccggt ccaggatctc agggaggtga acaagcgcgt cgaggatatc
     4201 catccgaccg tgccgaaccc atacaatctc ctgtcaggcc tcccgccatc tcaccagtgg
     4261 tacaccgtcc tcgacctgaa ggatgcgttc ttctgcctca ggctgcatcc aacaagccag
     4321 cctctcttcg ccttcgagtg gcgcgatcca gagatgggca tttcaggcca gctcacctgg
     4381 acacggctgc cacagggctt caagaactct cctaccctct tcaatgaggc gctccatcgg
     4441 gacctggccg atttcaggat ccagcaccct gacctcattc tcctgcagta cgtggacgat
     4501 ctcctgctcg ccgcgacatc agagctggat tgccagcagg gcaccagggc cctgctccag
     4561 acactcggca atctgggcta ccgggcctct gcgaagaagg cccagatctg ccagaagcag
     4621 gtgaagtacc tcggctacct gctcaaggag ggacagaggt ggctgacaga ggcaaggaag
     4681 gagacagtca tgggccagcc taccccgaag acacctcggc agctcaggga gttcctgggc
     4741 aaggccggat tctgcaggct cttcattcca ggattcgcgg agatggcggc gccactctac
     4801 cctctgacca agccgggcac actgttcaac tggggcccag accagcagaa ggcgtaccag
     4861 gagattaagc aggcactgct cacagcacct gcgctcggcc tgccggacct cacaaagcca
     4921 ttcgagctgt tcgtggatga gaagcagggc tacgcgaagg gagtcctgac acagaagctg
     4981 ggaccatgga ggcgcccagt ggcctacctc tcaaagaagc tcgacccagt ggcggccgga
     5041 tggcctccgt gcctgaggat ggtggcggcc attgccgtcc tcaccaagga tgccggcaag
     5101 ctgacaatgg gccagcctct cgtgattctg gcgccgcatg cggtggaggc cctggtcaag
     5161 cagccacctg ataggtggct gtccaacgcg cgcatgaccc actaccaggc cctgctcctg
     5221 gacacagata gggtccagtt cggaccagtg gtggcactca atcctgccac actgctgcca
     5281 ctccctgagg agggcctgca gcataactgc ctcgatattc tggcggaggc ccatggcacc
     5341 cggccagacc tcacagatca gccgctgcca gacgccgatc acacctggta cacagatggc
     5401 tcatctctcc tgcaggaggg ccagaggaag gccggagcag ccgtgaccac agagacagag
     5461 gtcatctggg caaaggccct cccagcgggc acctcagcac agagggccga gctcattgca
     5521 ctgacacagg cgctcaagat ggccgagggc aagaagctga atgtgtacac agactccagg
     5581 tacgcattcg ccacagcaca catccatggc gagatttaca ggcggagggg atggctcaca
     5641 tcagagggaa aggagatcaa gaacaaggat gagattctcg cgctcctgaa ggccctcttc
     5701 ctgcctaagc gcctgtcaat cattcactgc ccaggacatc agaagggaca ctcagccgag
     5761 gcaaggggaa ataggatggc agaccaggcg gccaggaagg cagcgatcac cgagacacca
     5821 gatacctcca cactcctgat tgagaactcc agccctagcg gtggctccag cggtggtagc
     5881 agcggtagcg aaactccagg gacctcggaa tcggcgactc cagaatccag tgggggtagc
     5941 agcggcggat ccagcgacaa gaagtactcc atcggcctcg acatcggcac caacagcgtc
     6001 ggctgggcgg tgatcaccga cgagtacaag gtcccgtcca agaagttcaa ggtcctgggc
     6061 aacaccgacc gccactccat caagaagaac ctcatcggcg ccctcctctt cgactccggc
     6121 gagacggcgg aggcgacccg cctcaagcgc accgcccgcc gccgctacac ccgccgcaag
     6181 aaccgcatct gctacctcca ggagatcttc tccaacgaga tggcgaaggt cgacgactcc
     6241 ttcttccacc gcctcgagga gtccttcctc gtggaggagg acaagaagca cgagcgccac
     6301 cccatcttcg gcaacatcgt cgacgaggtc gcctaccacg agaagtaccc cactatctac
     6361 caccttcgta agaagcttgt tgactctact gataaggctg atcttcgtct catctacctt
     6421 gctctcgctc acatgatcaa gttccgtggt cacttcctta tcgagggtga ccttaaccct
     6481 gataactccg acgtggacaa gctcttcatc cagctcgtcc agacctacaa ccagctcttc
     6541 gaggagaacc ctatcaacgc ttccggtgtc gacgctaagg cgatcctttc cgctaggctc
     6601 tccaagtcca ggcgtctcga gaacctcatc gcccagctcc ctggtgagaa gaagaacggt
     6661 cttttcggta acctcatcgc tctctccctc ggtctgaccc ctaacttcaa gtccaacttc
     6721 gacctcgctg aggacgctaa gcttcagctc tccaaggata cctacgacga tgatctcgac
     6781 aacctcctcg ctcagattgg agatcagtac gctgatctct tccttgctgc taagaacctc
     6841 tccgatgcta tcctcctttc ggatatcctt agggttaaca ctgagatcac taaggctcct
     6901 ctttctgctt ccatgatcaa gcgctacgac gagcaccacc aggacctcac cctcctcaag
     6961 gctcttgttc gtcagcagct ccccgagaag tacaaggaga tcttcttcga ccagtccaag
     7021 aacggctacg ccggttacat tgacggtgga gctagccagg aggagttcta caagttcatc
     7081 aagccaatcc ttgagaagat ggatggtact gaggagcttc tcgttaagct taaccgtgag
     7141 gacctcctta ggaagcagag gactttcgat aacggctcta tccctcacca gatccacctt
     7201 ggtgagcttc acgccatcct tcgtaggcag gaggacttct accctttcct caaggacaac
     7261 cgtgagaaga tcgagaagat ccttactttc cgtattcctt actacgttgg tcctcttgct
     7321 cgtggtaact cccgtttcgc ttggatgact aggaagtccg aggagactat caccccttgg
     7381 aacttcgagg aggttgttga caagggtgct tccgcccagt ccttcatcga gcgcatgacc
     7441 aacttcgaca agaacctccc caacgagaag gtcctcccca agcactccct cctctacgag
     7501 tacttcacgg tctacaacga gctcaccaag gtcaagtacg tcaccgaggg tatgcgcaag
     7561 cctgccttcc tctccggcga gcagaagaag gctatcgttg acctcctctt caagaccaac
     7621 cgcaaggtca ccgtcaagca gctcaaggag gactacttca agaagatcga gtgcttcgac
     7681 tccgtcgaga tcagcggcgt tgaggaccgt ttcaacgctt ctctcggtac ctaccacgat
     7741 ctcctcaaga tcatcaagga caaggacttc ctcgacaacg aggagaacga ggacatcctc
     7801 gaggacatcg tcctcactct tactctcttc gaggataggg agatgatcga ggagaggctc
     7861 aagacttacg ctcatctctt cgatgacaag gttatgaagc agctcaagcg tcgccgttac
     7921 accggttggg gtaggctctc ccgcaagctc atcaacggta tcagggataa gcagagcggc
     7981 aagactatcc tcgacttcct caagtctgat ggtttcgcta acaggaactt catgcagctc
     8041 atccacgatg actctcttac cttcaaggag gatattcaga aggctcaggt gtccggtcag
     8101 ggcgactctc tccacgagca cattgctaac cttgctggtt cccctgctat caagaagggc
     8161 atccttcaga ctgttaaggt tgtcgatgag cttgtcaagg ttatgggtcg tcacaagcct
     8221 gagaacatcg tcatcgagat ggctcgtgag aaccagacta cccagaaggg tcagaagaac
     8281 tcgagggagc gcatgaagag gattgaggag ggtatcaagg agcttggttc tcagatcctt
     8341 aaggagcacc ctgtcgagaa cacccagctc cagaacgaga agctctacct ctactacctc
     8401 cagaacggta gggatatgta cgttgaccag gagctcgaca tcaacaggct ttctgactac
     8461 gacgtcgacg ccattgttcc tcagtctttc cttaaggatg actccatcga caacaaggtc
     8521 ctcacgaggt ccgacaagaa caggggtaag tcggacaacg tcccttccga ggaggttgtc
     8581 aagaagatga agaactactg gaggcagctt ctcaacgcta agctcattac ccagaggaag
     8641 ttcgacaacc tcacgaaggc tgagaggggt ggcctttccg agcttgacaa ggctggtttc
     8701 atcaagaggc agcttgttga gacgaggcag attaccaagc acgttgctca gatcctcgat
     8761 tctaggatga acaccaagta cgacgagaac gacaagctca tccgcgaggt caaggtgatc
     8821 accctcaagt ccaagctcgt ctccgacttc cgcaaggact tccagttcta caaggtccgc
     8881 gagatcaaca actaccacca cgctcacgat gcttacctta acgctgtcgt tggtaccgct
     8941 cttatcaaga agtaccctaa gcttgagtcc gagttcgtct acggtgacta caaggtctac
     9001 gacgttcgta agatgatcgc caagtccgag caggagatcg gcaaggccac cgccaagtac
     9061 ttcttctact ccaacatcat gaacttcttc aagaccgaga tcaccctcgc caacggcgag
     9121 atccgcaagc gccctcttat cgagacgaac ggtgagactg gtgagatcgt ttgggacaag
     9181 ggtcgcgact tcgctactgt tcgcaaggtc ctttctatgc ctcaggttaa catcgtcaag
     9241 aagaccgagg tccagaccgg tggcttctcc aaggagtcta tccttccaaa gagaaactcg
     9301 gacaagctca tcgctaggaa gaaggattgg gaccctaaga agtacggtgg tttcgactcc
     9361 cctactgtcg cctactccgt cctcgtggtc gccaaggtgg agaagggtaa gtcgaagaag
     9421 ctcaagtccg tcaaggagct cctcggcatc accatcatgg agcgctcctc cttcgagaag
     9481 aacccgatcg acttcctcga ggccaagggc tacaaggagg tcaagaagga cctcatcatc
     9541 aagctcccca agtactctct tttcgagctc gagaacggtc gtaagaggat gctggcttcc
     9601 gctggtgagc tccagaaggg taacgagctt gctcttcctt ccaagtacgt gaacttcctc
     9661 tacctcgcct cccactacga gaagctcaag ggttcccctg aggataacga gcagaagcag
     9721 ctcttcgtgg agcagcacaa gcactacctc gacgagatca tcgagcagat ctccgagttc
     9781 tccaagcgcg tcatcctcgc tgacgctaac ctcgacaagg tcctctccgc ctacaacaag
     9841 caccgcgaca agcccatccg cgagcaggcc gagaacatca tccacctctt cacgctcacg
     9901 aacctcggcg cccctgctgc tttcaagtac ttcgacacca ccatcgacag gaagcgttac
     9961 acgtccacca aggaggttct cgacgctact ctcatccacc agtccatcac cggtctttac
    10021 gagactcgta tcgacctttc ccagcttggt ggtgatgacg atgacaaaat ggcaccgaag
    10081 aaaaaaagga aggtcggcgg ctccccgaag aaaaaaagga aggtcggcgg ctccccgaag
    10141 aaaaaaagga aggtcggcgg ctccccgaag aaaaaaagga aggtcggaat ccatggcgga
    10201 tcaggagcca ccaacttctc cctcctcaag caggccggcg acgtggagga gaacccgggc
    10261 ccaatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
    10321 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
    10381 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
    10441 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
    10501 attggggagt ttagcgagag cctgacctat tgcatctccc gccgttcaca gggtgtcacg
    10561 ttgcaagacc tgcctgaaac cgaactgccc gctgttctac aaccggtcgc ggaggctatg
    10621 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
    10681 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
    10741 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
    10801 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
    10861 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
    10921 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
    10981 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
    11041 ccacgactcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
    11101 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
    11161 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
    11221 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
    11281 aagaaataga ctagttcccg atcgttcaaa catttggcaa taaagtttct taagattgaa
    11341 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
    11401 aataattaac atgtaatgca tgacgttatt tatgaggtgg gtttttatga ttagagtccc
    11461 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
    11521 atcgcgcgcg gtgtcatcta tgttactaga cctgcaggtg gaatcggcag caaaggattt
    11581 tttcctgtag ttttcccaca accatttttt accatccgaa tgataggata ggaaaaatat
    11641 ccaagtgaac agtattccta taaaattccc gtaaaaagcc tgcaatccga atgagccctg
    11701 aagtctgaac tagccggtca cctgtacagg ctatcgagat gccatacaag agacggtagt
    11761 aggaactagg aagacgatgg ttgattcgtc aggcgaaatc gtcgtcctgc agtcgcatct
    11821 atgggcctgg acggaatagg ggaaaaagtt ggccggatag gagggaaagg cccaggtgct
    11881 tacgtgcgag gtaggcctgg gctctcagca cttcgattcg ttggcaccgg ggtaggatgc
    11941 aatagagagc aacgtttagt accacctcgc ttagctagag caaactggac tgccttatat
    12001 gcgcgggtgc tggcttggct gccgtgagac cgaggtctcg ttttttttaa gcttggcgag
    12061 ctagcggccg catgcatcga tctcctacat cgtataaatt agcctatacg aagttattgc
    12121 atctatgtcg ggtgcggaga aagaggtaat gaaatggcag tattagatct gataacttcg
    12181 tataatgtat gctatacgaa gttatgactg caggtcgaca cccataatag ctgtttgcca
    12241 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
    12301 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
    12361 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc
    12421 ttggatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat
    12481 tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg
    12541 tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg
    12601 gatcaaagta ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca
    12661 gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc
    12721 gccctgccct tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact
    12781 tgcgactaga accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc
    12841 tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg
    12901 gccggctgca ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag
    12961 ctggccagga tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac
    13021 cgcctggccc gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc
    13081 gcgggcctgc gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg
    13141 gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc
    13201 cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc
    13261 accccggcac agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa
    13321 gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc
    13381 gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc
    13441 gaggccgacg ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc
    13501 accaggacgg ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg
    13561 cggccgggta cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc
    13621 tggccggttt gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa
    13681 ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg
    13741 tcgctgcgta tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt
    13801 tatcgctgta cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc
    13861 ccgcgccctg caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag
    13921 tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg
    13981 cccgacgatt gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg
    14041 agcgccccag gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat
    14101 tccggtgcag ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa
    14161 gcagcgcatt gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat
    14221 caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat
    14281 tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac
    14341 cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga
    14401 aattaaatca aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac
    14461 acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg
    14521 gcagacacgc cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag
    14581 ctgaagatgt acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc
    14641 gcgcagctac cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa
    14701 aggaggcggc atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg
    14761 tggaggaacg ggcggttggc caggcgtaag cggctgggtt gcctgccggc cctgcaatgg
    14821 cactggaacc cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta
    14881 caaatcggcg cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc
    14941 cagcggcaac gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat
    15001 cgaatccgca aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg
    15061 cccaagggcg acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc
    15121 gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct
    15181 ggcgaggtga tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc
    15241 ggcatggcca gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa
    15301 tccatgaacc gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac
    15361 gttgcggacg tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg
    15421 gtagaaacct gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc
    15481 aagaacggcc gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc
    15541 gtaaagagcg aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac
    15601 cgcgagatca cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg
    15661 atcgatcccg gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca
    15721 gaagccagat ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag
    15781 aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg
    15841 aaggaggagg cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag
    15901 ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca
    15961 ggggaaaaag gtcgaaaagt tctctttcct gtggatagca cgtacattgg gaacccaaag
    16021 ccgtacattg ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg
    16081 tcacacatgt aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct
    16141 ttaaaactta ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc
    16201 acagccgaag agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc
    16261 gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc
    16321 aatctaccag ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa
    16381 ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
    16441 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
    16501 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
    16561 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
    16621 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
    16681 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    16741 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    16801 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    16861 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    16921 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    16981 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    17041 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    17101 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    17161 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    17221 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    17281 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    17341 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    17401 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    17461 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat
    17521 tctaggtact aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc
    17581 ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg
    17641 atcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga
    17701 tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg
    17761 tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc
    17821 ggatctttaa atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta
    17881 ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc
    17941 gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca
    18001 gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc
    18061 agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct
    18121 tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg
    18181 ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata taccttagca
    18241 ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc
    18301 ggtgatattc tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga
    18361 taccccaaga agctaaggaa ggtgcgaaca agtccctgat atgagatc
//
