LOCUS       sequence-410941-       19402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NEPE for prime editing in rice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19402)
  AUTHORS   Zhong Z, Fan T, He Y, Liu S, Zheng X, Xu Y, Ren J, Yuan H, Xu Z, 
            Zhang Y
  TITLE     An improved plant prime editor for efficient generation of 
            multiple-nucleotide variations and structural variations in rice.
  JOURNAL   Plant Commun. 2024 May 14:100976. doi: 10.1016/j.xplc.2024.100976.
  PUBMED    38751122
REFERENCE   2  (bases 1 to 19402)
  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 
            Commun. 2024 May 14:100976. doi: 10.1016/j.xplc.2024.100976."
COMMENT     Sequence Label: pGEL685
FEATURES             Location/Qualifiers
     source          1..19402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2006
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             2022..2069
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4242..4262
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4263..6779,6780..6782,6783..8366)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     RBS             8279..8287
                     /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             8385..8405
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8418..8438
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8451..8471
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8480..8728
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     CDS             complement(8990..9295)
                     /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     complement(9092..9111)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(9626..9650)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    9680..9704
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11005..11634
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             12063..13136
                     /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      13202..13396
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13632..13654)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13730..13870
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13785..13804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13885..13902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14056..14644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14136..14155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(16069..16863)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    17288..17312
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    17390..17564
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        complement(18654..19330)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(18697..18717)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 ggcgcgcctg cagtgcagcg tgacccggtc gtgcccctct cttgagataa ggtgagcatt
       61 gcatgtctaa gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca
      121 gtttatctat ctttatacat atatttaaac tttactctac gaataatata atctatagta
      181 ctacaataat atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag
      241 gacaattgag tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt
      301 ctcctttttt tttgcaaata gcttcaccta tataatactt catccatttt attagtacat
      361 ccatttaggg tttagggtta atggttttta tagactaatt tttttagtac atctatttta
      421 ttctatttta gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata
      481 atttagatat aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga
      541 aattaaaaaa actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa
      601 cgccgtcgac gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag
      661 cgaagcagac ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc
      721 caccgttgga cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg
      781 agccggcacg gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc
      841 tttcccaccg ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc
      901 acaccctctt tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagaactccc
      961 ccaaatccac ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc cccccccccc
     1021 ctctctacct tctcaagatc ggcgttccgg tccatggtta gggcccggta gttctacttc
     1081 tgttcatgtt tgtgttagat ccgtgtttgt gttagatccg tgctactagc gttcgtacac
     1141 ggatgcgacc tgtacgtcag acacgttctg attgctaact tgccagtgtt tctctttggg
     1201 gaatcctggg atggctctag ccgttccgca gacgggatcg atttcatgat tttttttgtt
     1261 tcgttgcata gggtttggtt tgcccttttc ctttatttca atatatgccg tgcacttgtt
     1321 tgtcgggtca tcttttcatg cttttttttg tcttggttgt gatgatgtgg tctggttggg
     1381 cggtcgttca agatcggagt agaattaatt ctgtttcaaa ctacctggtg gatttattaa
     1441 ttttggatct gtatgtgtgt gccatacata ttcatagtta cgaattgaag atgatggatg
     1501 gaaatatcga tctaggatag gtatacatgt tgatgcgggt tttactgatg catatacaga
     1561 gatgcttttt gttcgcttgg ttgtgatgat gtggtgtggt tgggcggtcg ttcattcgtt
     1621 caagatcgga gtagaatact gtttcaaact acctggtgta tttattaatt ttggaactgt
     1681 atgtgtgtgt catacatctt catagttacg agtttaagat ggatggaaat atcgatctag
     1741 gataggtata catgttgatg tgggttttac tgatgcatat acatgatggc atatgcagca
     1801 tctattcata tgctctaacc ttgagtacct atctattata ataaacaagt atgttttata
     1861 attattttga tcttgatata cttggatgat ggcatatgca gcagctatat gtggattttt
     1921 ttagccctgc cttcatacgc tatttatttg cttggtactg tttcttttgt cgatgctcac
     1981 cctgttgttt ggtgttactt ctgcagcctg caggccatat gaagcggcca gcggcgacga
     2041 agaaggcggg gcaggcgaag aagaagaagt cagggggttc tagcggcggt tcatctggct
     2101 ccgaaacacc gggaacttcc gagagtgcta ctccagaatc tagcggtggg tcatctggag
     2161 ggtcttccac acttaatatt gaggacgaat accgcctgca tgaaactagt aaagaacccg
     2221 acgtgagcct cggtagcacg tggctctctg atttcccgca agcatgggct gaaacaggtg
     2281 ggatgggcct cgctgttcgc caggctccat tgatcatccc actcaaggcc acttctacac
     2341 cggtgtccat aaagcaatac ccaatgagcc aggaagctcg gcttggtatc aagcctcaca
     2401 tccaaagatt gcttgatcag ggaattcttg tcccttgtca aagtccatgg aacacccctc
     2461 ttcttcccgt caaaaaacct gggactaacg attataggcc agtgcaagac ctcagggagg
     2521 ttaacaaaag ggttgaagac attcatccaa cagtgccgaa tccttataat ctgctctcag
     2581 gtttgcctcc aagtcaccag tggtatacag tgctggatct taaagacgca tttttttgtc
     2641 tgcgccttca tcccacttcg caacctcttt ttgctttcga atggagagat ccagagatgg
     2701 gtatctccgg acagctcaca tggacccggt tgccccaagg cttcaaaaac tccccaacct
     2761 tgtttaatga agcgctgcac agagatttgg cggacttcag gatccagcac cccgacttga
     2821 ttctcctgca gtatgtggat gaccttttgc ttgcagccac atctgagctt gactgccaac
     2881 aggggacgcg cgcccttctt cagactttgg gaaaccttgg gtatagagcc tctgccaaga
     2941 aggcacagat ctgccagaag caagttaagt atctcggtta tttgctcaag gagggtcaga
     3001 ggtggctgac ggaggcgagg aaagaaacgg tgatgggcca accaacccca aaaaccccta
     3061 ggcaacttag agagtttctt ggtaaagcgg ggttttgccg gttgttcatt ccaggctttg
     3121 ccgagatggc cgcaccgttg tatccattga cgaaaccggg cacccttttt aattgggggc
     3181 ccgaccagca gaaggcgtat caggagatca agcaagctct cctgaccgca ccagcactcg
     3241 gacttccaga tcttacgaaa ccgtttgagc tcttcgtgga tgagaaacag gggtatgcta
     3301 agggagtcct cacacaaaag ttgggaccgt ggaggcggcc ggttgcctat ctttcgaaaa
     3361 aacttgatcc cgtggcggct ggttggcctc cctgtctgag gatggtggcc gcgatcgcag
     3421 tccttacaaa agatgcaggg aagttgacta tgggacaacc attggtcata cttgccccgc
     3481 acgcggttga ggccctcgtt aagcagcctc ccgataggtg gctcagtaac gcccgcatga
     3541 ctcattacca ggcactgctt ctcgacacag acagggtgca gtttggtccc gtcgtcgctt
     3601 tgaaccccgc caccctgctg cccctcccgg aggaaggatt gcagcacaac tgtctggata
     3661 tcttggccga agcgcatggg actcgcccgg acttgactga ccagcccctt cccgacgctg
     3721 accatacttg gtatacggat ggaagcagcc ttctgcagga agggcaacgc aaggcagggg
     3781 ccgcggttac tactgaaact gaggtcatct gggctaaggc tttgccagcc ggaactagtg
     3841 ctcaaagagc agaattgatt gccctcacgc aagctttgaa aatggctgag ggtaagaagc
     3901 ttaatgtgta tacagacagt aggtatgcat ttgcgacagc acatatccat ggtgagatct
     3961 accggcgcag aggatggctt acgagcgagg ggaaggagat taagaacaaa gatgagattt
     4021 tggccctgct caaagcgctc ttcctcccga agcgccttag catcatccat tgcccaggcc
     4081 atcagaaggg tcactcggcc gaagcacggg gtaataggat ggccgatcag gcagcaagga
     4141 aggctgctat cactgagaca cccgacacct ctacccttct catagagaat tcatcgccga
     4201 gcggcggcag caagaggact gcagacgggt cggaattcga accgaagaaa aagagaaagg
     4261 taatggacaa gaagtactcg atcggcctcg atattgggac taactctgtt ggctgggccg
     4321 tgatcaccga cgagtacaag gtgccctcaa agaagttcaa ggtcctgggc aacaccgatc
     4381 ggcattccat caagaagaat ctcattggcg ctctcctgtt cgacagcggc gagacggctg
     4441 aggctacgcg gctcaagcgc accgcccgca ggcggtacac gcgcaggaag aatcgcatct
     4501 gctacctgca ggagattttc tccaacgaga tggcgaaggt tgacgattct ttcttccaca
     4561 ggctggagga gtcattcctc gtggaggagg ataagaagca cgagcggcat ccaatcttcg
     4621 gcaacattgt cgacgaggtt gcctaccacg agaagtaccc tacgatctac catctgcgga
     4681 agaagctcgt ggactccaca gataaggcgg acctccgcct gatctacctc gctctggccc
     4741 acatgattaa gttcaggggc catttcctga tcgaggggga tctcaacccg gacaatagcg
     4801 atgttgacaa gctgttcatc cagctcgtgc agacgtacaa ccagctcttc gaggagaacc
     4861 ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc cgctaggctc tcgaagtctc
     4921 ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa gaagaacggc ctgttcggga
     4981 atctcattgc gctcagcctg gggctcacgc ccaacttcaa gtcgaatttc gatctcgctg
     5041 aggacgccaa gctgcagctc tccaaggaca catacgacga tgacctggat aacctcctgg
     5101 cccagatcgg cgatcagtac gcggacctgt tcctcgctgc caagaatctg tcggacgcca
     5161 tcctcctgtc tgatattctc agggtgaaca ccgagattac gaaggctccg ctctcagcct
     5221 ccatgatcaa gcgctacgac gagcaccatc aggatctgac cctcctgaag gcgctggtca
     5281 ggcagcagct ccccgagaag tacaaggaga tcttcttcga tcagtcgaag aacggctacg
     5341 ctgggtacat tgacggcggg gcctctcagg aggagttcta caagttcatc aagccgattc
     5401 tggagaagat ggacggcacg gaggagctgc tggtgaagct caatcgcgag gacctcctga
     5461 ggaagcagcg gacattcgat aacggcagca tcccacacca gattcatctc ggggagctgc
     5521 acgctatcct gaggaggcag gaggacttct accctttcct caaggataac cgcgagaaga
     5581 tcgagaagat tctgactttc aggatcccgt actacgtcgg cccactcgct aggggcaact
     5641 cccgcttcgc ttggatgacc cgcaagtcag aggagacgat cacgccgtgg aacttcgagg
     5701 aggtggtcga caagggcgct agcgctcagt cgttcatcga gaggatgacg aatttcgaca
     5761 agaacctgcc aaatgagaag gtgctcccta agcactcgct cctgtacgag tacttcacag
     5821 tctacaacga gctgactaag gtgaagtatg tgaccgaggg catgaggaag ccggctttcc
     5881 tgtctgggga gcagaagaag gccatcgtgg acctcctgtt caagaccaac cggaaggtca
     5941 cggttaagca gctcaaggag gactacttca agaagattga gtgcttcgat tcggtcgaga
     6001 tctctggcgt tgaggaccgc ttcaacgcct ccctggggac ctaccacgat ctcctgaaga
     6061 tcattaagga taaggacttc ctggacaacg aggagaatga ggatatcctc gaggacattg
     6121 tgctgacact cactctgttc gaggaccggg agatgatcga ggagcgcctg aagacttacg
     6181 cccatctctt cgatgacaag gtcatgaagc agctcaagag gaggaggtac accggctggg
     6241 ggaggctgag caggaagctc atcaacggca ttcgggacaa gcagtccggg aagacgatcc
     6301 tcgacttcct gaagagcgat ggcttcgcga accgcaattt catgcagctg attcacgatg
     6361 acagcctcac attcaaggag gatatccaga aggctcaggt gagcggccag ggggactcgc
     6421 tgcacgagca tatcgcgaac ctcgctggct cgccagctat caagaagggg attctgcaga
     6481 ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag gcacaagcct gagaacatcg
     6541 tcattgagat ggcccgggag aatcagacca cgcagaaggg ccagaagaac tcacgcgaga
     6601 ggatgaagag gatcgaggag ggcattaagg agctggggtc ccagatcctc aaggagcacc
     6661 cggtggagaa cacgcagctg cagaatgaga agctctacct gtactacctc cagaatggcc
     6721 gcgatatgta tgtggaccag gagctggata ttaacaggct cagcgattac gacgtcgatg
     6781 ctatcgttcc acagtcattc ctgaaggatg actccattga caacaaggtc ctcaccaggt
     6841 cggacaagaa ccggggcaag tctgataatg ttccttcaga ggaggtcgtt aagaagatga
     6901 agaactactg gcgccagctc ctgaatgcca agctgatcac gcagcggaag ttcgataacc
     6961 tcacaaaggc tgagaggggc gggctctctg agctggacaa ggcgggcttc atcaagaggc
     7021 agctggtcga gacacggcag atcactaagc acgttgcgca gattctcgac tcacggatga
     7081 acactaagta cgatgagaat gacaagctga tccgcgaggt gaaggtcatc accctgaagt
     7141 caaagctcgt ctccgacttc aggaaggatt tccagttcta caaggttcgg gagatcaaca
     7201 attaccacca tgcccatgac gcgtacctga acgcggtggt cggcacagct ctgatcaaga
     7261 agtacccaaa gctcgagagc gagttcgtgt acggggacta caaggtttac gatgtgagga
     7321 agatgatcgc caagtcggag caggagattg gcaaggctac cgccaagtac ttcttctact
     7381 ctaacattat gaatttcttc aagacagaga tcactctggc caatggcgag atccggaagc
     7441 gccccctcat cgagacgaac ggcgagacgg gggagatcgt gtgggacaag ggcagggatt
     7501 tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa tatcgtcaag aagacagagg
     7561 tccagactgg cgggttctct aaggagtcaa ttctgcctaa gcggaacagc gacaagctca
     7621 tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg gttcgacagc cccactgtgg
     7681 cctactcggt cctggttgtg gcgaaggttg agaagggcaa gtccaagaag ctcaagagcg
     7741 tgaaggagct gctggggatc acgattatgg agcgctccag cttcgagaag aacccgatcg
     7801 atttcctgga ggcgaagggc tacaaggagg tgaagaagga cctgatcatt aagctcccca
     7861 agtactcact cttcgagctg gagaacggca ggaagcggat gctggcttcc gctggcgagc
     7921 tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt gaacttcctc tacctggcct
     7981 cccactacga gaagctcaag ggcagccccg aggacaacga gcagaagcag ctgttcgtcg
     8041 agcagcacaa gcattacctc gacgagatca ttgagcagat ttccgagttc tccaagcgcg
     8101 tgatcctggc cgacgcgaat ctggataagg tcctctccgc gtacaacaag caccgcgaca
     8161 agccaatcag ggagcaggct gagaatatca ttcatctctt caccctgacg aacctcggcg
     8221 cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg caagaggtac acaagcacta
     8281 aggaggtcct ggacgcgacc ctcatccacc agtcgattac cggcctctac gagacgcgca
     8341 tcgacctgtc tcagctcggg ggcgacacta gttccggcgg cagcccaaag aagaagcgga
     8401 aggtgtctgg aggttctcct aagaaaaaga gaaaagtgtc cggcggctcc ccgaagaaga
     8461 agcgcaaggt gtgagcttat atgaagatga agatgaaata tttggtgtgt caaataaaaa
     8521 gcttgtgtgc ttaagtttgt gtttttttct tggcttgttg tgttatgaat ttgtggcttt
     8581 ttctaatatt aaatgaatgt aagatcacat tataatgaat aaacaaatgt ttctataatc
     8641 cattgtgaat gttttgttgg atctcttctg cagcatataa ctactgtatg tgctatggta
     8701 tggactatgg aatatgatta aagataagag gatccgcgga tcatgaacca acggcctggc
     8761 tgtatttggt ggttgtgtag ggagatgggg agaagaaaag cccgattctc ttcgctgtga
     8821 tgggctggat gcatgcgggg gagcgggagg cccaagtacg tgcacggtga gcggcccaca
     8881 gggcgagtgt gagcgcgaga ggcgggagga acagtttagt accacattgc ccagctaact
     8941 cgaacgcgac caacttataa acccgcgcgc tgtcgcttgt gtggagacct tatattcccc
     9001 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
     9061 cttccccgat aacggaaacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
     9121 catccccgat atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg
     9181 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
     9241 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
     9301 cagcccctgt tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
     9361 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
     9421 atggttgtgc ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc
     9481 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
     9541 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
     9601 ctgttatctg gcttggtctc tctagaccca gctttcttgt acaaagttgg cattactgcg
     9661 agctcaaact atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg
     9721 tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg
     9781 tatgtgcatg ccaaccacag ggttcccctc gggatcaaag tactttgatc caacccctcc
     9841 gctgctatag tgcagtcggc ttctgacgtt cagtgcagcc gtcttctgaa aacgacatgt
     9901 cgcacaagtc ctaagttacg cgacaggctg ccgccctgcc cttttcctgg cgttttcttg
     9961 tcgcgtgttt tagtcgcata aagtagaata cttgcgacta gaaccggaga cattacgcca
    10021 tgaacaagag cgccgccgct ggcctgctgg gctatgcccg cgtcagcacc gacgaccagg
    10081 acttgaccaa ccaacgggcc gaactgcacg cggccggctg caccaagctg ttttccgaga
    10141 agatcaccgg caccaggcgc gaccgcccgg agctggccag gatgcttgac cacctacgcc
    10201 ctggcgacgt tgtgacagtg accaggctag accgcctggc ccgcagcacc cgcgacctac
    10261 tggacattgc cgagcgcatc caggaggccg gcgcgggcct gcgtagcctg gcagagccgt
    10321 gggccgacac caccacgccg gccggccgca tggtgttgac cgtgttcgcc ggcattgccg
    10381 agttcgagcg ttccctaatc atcgaccgca cccggagcgg gcgcgaggcc gccaaggccc
    10441 gaggcgtgaa gtttggcccc cgccctaccc tcaccccggc acagatcgcg cacgcccgcg
    10501 agctgatcga ccaggaaggc cgcaccgtga aagaggcggc tgcactgctt ggcgtgcatc
    10561 gctcgaccct gtaccgcgca cttgagcgca gcgaggaagt gacgcccacc gaggccaggc
    10621 ggcgcggtgc cttccgtgag gacgcattga ccgaggccga cgccctggcg gccgccgaga
    10681 atgaacgcca agaggaacaa gcatgaaacc gcaccaggac ggccaggacg aaccgttttt
    10741 cattaccgaa gagatcgagg cggagatgat cgcggccggg tacgtgttcg agccgcccgc
    10801 gcacggctca accgtgcggc tgcatgaaat cctggccggt ttgtctgatg ccaagctggc
    10861 ggcctggccg gccagcttgg ccgctgaaga aaccgagcgc cgccgtctaa aaaggtgatg
    10921 tgtatttgag taaaacagct tgcgtcatgc ggtcgctgcg tatatgatgc gatgagtaaa
    10981 taaacaaata cgcaagggga acgcatgaag gttatcgctg tacttaacca gaaaggcggg
    11041 tcaggcaaga cgaccatcgc aacccatcta gcccgcgccc tgcaactcgc cggggccgat
    11101 gttctgttag tcgattccga tccccagggc agtgcccgcg attgggcggc cgtgcgggaa
    11161 gatcaaccgc taaccgttgt cggcatcgac cgcccgacga ttgaccgcga cgtgaaggcc
    11221 atcggccggc gcgacttcgt agtgatcgac ggagcgcccc aggcggcgga cttggctgtg
    11281 tccgcgatca aggcagccga cttcgtgctg attccggtgc agccaagccc ttacgacata
    11341 tgggccaccg ccgacctggt ggagctggtt aagcagcgca ttgaggtcac ggatggaagg
    11401 ctacaagcgg cctttgtcgt gtcgcgggcg atcaaaggca cgcgcatcgg cggtgaggtt
    11461 gccgaggcgc tggccgggta cgagctgccc attcttgagt cccgtatcac gcagcgcgtg
    11521 agctacccag gcactgccgc cgccggcaca accgttcttg aatcagaacc cgagggcgac
    11581 gctgcccgcg aggtccaggc gctggccgct gaaattaaat caaaactcat ttgagttaat
    11641 gaggtaaaga gaaaatgagc aaaagcacaa acacgctaag tgccggccgt ccgagcgcac
    11701 gcagcagcaa ggctgcaacg ttggccagcc tggcagacac gccagccatg aagcgggtca
    11761 actttcagtt gccggcggag gatcacacca agctgaagat gtacgcggta cgccaaggca
    11821 agaccattac cgagctgcta tctgaataca tcgcgcagct accagagtaa atgagcaaat
    11881 gaataaatga gtagatgaat tttagcggct aaaggaggcg gcatggaaaa tcaagaacaa
    11941 ccaggcaccg acgccgtgga atgccccatg tgtggaggaa cgggcggttg gccaggcgta
    12001 agcggctggg ttgcctgccg gccctgcaat ggcactggaa cccccaagcc cgaggaatcg
    12061 gcgtgagcgg tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc
    12121 tggtggagaa gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac
    12181 gccccggtga atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc
    12241 cggcagccgg tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt
    12301 tcgttccgat gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg
    12361 ttttccgtct gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag
    12421 acgggcacgt agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc
    12481 tggtactgat ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg
    12541 gagacaagcc cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc
    12601 gagccgatgg cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca
    12661 cgcacgttgc catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg
    12721 agggtgaagc cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt
    12781 acatcgagat cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg
    12841 acgtgctgac ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct
    12901 accgcctggc acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct
    12961 acgaacgcag tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga
    13021 tcgggtcaaa tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga
    13081 tcctagtcat gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta
    13141 cggagcagat gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggcctctttc
    13201 ctgtggatag cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca
    13261 ttgggaaccc aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag
    13321 agaaaaaagg cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc
    13381 gcctggcctg tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta
    13441 cccttcggtc gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg
    13501 gccgctcaaa aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc
    13561 cgtcgccact cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat
    13621 gacggtgaaa acctctgaca catgcagctc ccggaaacgg tcacagcttg tctgtaagcg
    13681 gatgccggga gcagacaagc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
    13741 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
    13801 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
    13861 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    13921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    13981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    14041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    14101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    14161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    14221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    14281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    14341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    14401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    14461 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
    14521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    14581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    14641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    14701 ttaagggctg atgaatcccc taatgatttt tatcaaaatc attaagttaa ggtagataca
    14761 catcttgtca tatgatcaaa tggtttcgcc aaaaatcaat aatcagacaa caaaatgtgc
    14821 gaactcgata ttttacacga ctctctttac caattctgcc ccgaattaca cttaaaacga
    14881 ctcaacagct taacgttggc ttgccacgcc ttacttgact gtaaaactct cactcttacc
    14941 gaacttggcc gtaacctgcc aaccaaagcg agaacaaaac ataacatcaa acgaatcgac
    15001 cgattgttag gtaatcgtca cctccacaaa gagcgactcg ctgtataccg ttggcatgct
    15061 agctttatct gttcgggcaa tacgatgccc attgtacttg ttgactggtc tgatatccgt
    15121 gagcaaaaac ggcttatggt attgcgagct tcagtcgcac tacacggtcg ttctgttact
    15181 ctttatgaga aagcgttccc gctttcagag caatgttcaa agaaagctca tgaccaattt
    15241 ctagccgacc ttgcgagcat tctaccgagt aacaccacac cgctcattgt cagtgatgct
    15301 ggctttaaag tgccatggta taaatccgtt gagaagctgg gttggtactg gttaagtcga
    15361 gtaagaggaa aagtacaata tgcagaccta ggagcggaaa actggaaacc tatcagcaac
    15421 ttacatgata tgtcatctag tcactcaaag actttaggct ataagaggct gactaaaagc
    15481 aatccaatct catgccaaat tctattgtat aaatctcgct ctaaaggccg aaaaaatcag
    15541 cgctcgacac ggactcattg tcaccacccg tcacctaaaa tctactcagc gtcggcaaag
    15601 gagccatgga ttctagcaac taacttacct gttgaaattc gaacacccaa acaacttgtt
    15661 aatatctatt cgaagcgaat gcagattgaa gaaaccttcc gagacttgaa aagtcctgcc
    15721 tacggactag gcctacgcca tagccgaacg agcagctcag agcgttttga tatcatgctg
    15781 ctaatcgccc tgatgcttca actaacatgt tggcttgcgg gcgttcatgc tcagaaacaa
    15841 ggttgggaca agcacttcca ggctaacaca gtcagaaatc gaaacgtact ctcaacagtt
    15901 cgcttaggca tggaagtttt gcggcattct ggctacacaa taacaaggga agacttactc
    15961 gtggctgcaa ccctactagc tcaaaattta ttcacacatg gttacgcttt ggggaaatta
    16021 tgaggggatc tctcagcgtt aagggatttt ggtcatgcat tctaggtact aaaacaattc
    16081 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
    16141 aaatagctcg acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc
    16201 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
    16261 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
    16321 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
    16381 ttcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
    16441 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
    16501 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
    16561 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
    16621 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
    16681 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
    16741 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
    16801 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
    16861 catttattat ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat
    16921 aacaagacga actccaattc actgttcctt gcattctaaa accttaaata ccagaaaaca
    16981 gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga gccgattttg
    17041 aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc
    17101 cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg
    17161 gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg
    17221 atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc
    17281 tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca
    17341 cattgcggac gtttttaatg tactgaatta acgccgaatt aattcggggg atctggattt
    17401 tagtactgga ttttggtttt aggaattaga aattttattg atagaagtat tttacaaata
    17461 caaatacata ctaagggttt cttatatgct caacacatga gcgaaaccct ataggaaccc
    17521 taattccctt atctgggaac tactcacaca ttattatgga gaaaatagag cttgtcgatc
    17581 gttaattaat cacctccaag atcccggacg agtgctgggg cgtcggtttc cactatcggc
    17641 gagtacttct acacagccat cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg
    17701 cccgacagtc ccggctccgg atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc
    17761 atcatcgaaa ttgccgtcaa ccaagctctg atagagttgg tcaagaccaa tgcggagcat
    17821 atacgcccgg agccgcggcg atcctgcaag ctccggatgc ctccgctcga agtagcgcgt
    17881 ctgctgctcc atacaagcca accacggcct ccagaagaag atgttggcga cctcgtattg
    17941 ggaatccccg aacatcgcct cgctccagtc aatgaccgct gttatgcggc cattgtccgt
    18001 caggacattg ttggagccga aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc
    18061 ggcccaaagc atcagctcat cgagagcctg cgcgacggac gcactgacgg tgtcgtccat
    18121 cacagtttgc cagtgataca catggggatc agcaatcgcg catatgaaat cacgccatgt
    18181 agtgtattga ccgattcctt gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc
    18241 ggccgcagcg atcgcatcca tggcctccgc gaccggctgc agaacagcgg gcagttcggt
    18301 ttcaggcagg tcttgcaacg tgacaccctg tgcacggcgg gagatgcaat aggtcaggct
    18361 ctcgctgaat tccccaatgt caagcacttc cggaatcggg agcgcggccg atgcaaagtg
    18421 ccgataaaca taacgatctt tgtagaaacc atcggcgcag ctatttaccc gcaggacata
    18481 tccacgccct cctacatcga agctgaaagc acgagattct tcgccctccg agagctgcat
    18541 caggtcggac acgctgtcga acttttcgat cagaaacttc tcgacagacg tcgcggtgag
    18601 ttcaggcttt ttcatcctag gagagataga tttgtagaga gagactggtg atttcagcgt
    18661 gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg
    18721 gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac
    18781 gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
    18841 ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg
    18901 taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg
    18961 aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct
    19021 tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgttc
    19081 acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct
    19141 cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt
    19201 tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat
    19261 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg
    19321 aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac
    19381 gagagtgtcg tgctccacca tg
//
