LOCUS       sequence-488644-       18589 bp    DNA     circular SYN 07-OCT-2025
DEFINITION  For plant prime editing including large DNA fragment editing in 
            Solanum lycopersicum or other dicotyledons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18589)
  AUTHORS   Zhao Y, Huang Z, Zhou X, Teng W, Liu Z, Wang W, Tang S, Liu Y, Liu 
            J, Wang W, Chai L, Zhang N, Guo W, Liu J, Ni Z, Sun Q, Wang Y, Zong 
            Y
  TITLE     Precise deletion, replacement and inversion of large DNA fragments 
            in plants using dual prime editing.
  JOURNAL   Nat Plants. 2025 Feb;11(2):191-205. doi: 10.1038/s41477-024-01898-3.
            Epub 2025 Jan 13.
  PUBMED    39805934
REFERENCE   2  (bases 1 to 18589)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41477-024-01898-3"; journalName: "Nat Plants"; date: 
            "2025-02"; volume: "11"; issue: "2"; pages: "191-205"
COMMENT     Sequence Label: pH-EF1-alpha-DualPE
FEATURES             Location/Qualifiers
     source          1..18589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    776..916
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(842..861)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1002..1024
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      1260..1454
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(1520..2593)
                     /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(3022..3651)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    4952..4976
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    5460..5479
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    5635..5654
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     polyA_signal    5661..5835
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             8000..8056
                     /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             8060..8086
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             8126..8146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             12160..12168
                     /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             12470..12490
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14216..14236
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(14895..14911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14895..14911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14908..14930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14919..14935
                     /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(14943..14949,14950..14967,14968..14973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14988..15009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        15200..15876
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     15813..15833
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             15943..16968
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    17009..17183
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    17261..17285
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             17710..18504
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
      781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      841 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
      901 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      961 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     1021 gaggttttca ccgtcatcac cgaaacgcgc gaggcagggt gccttgatgt gggcgccggc
     1081 ggtcgagtgg cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag
     1141 ccatttttga gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc
     1201 gcagcgaccg aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt
     1261 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
     1321 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
     1381 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
     1441 acgtgctatc cacaggaaac agaccttttc gacctttttc ccctgctagg gcaatttgcc
     1501 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
     1561 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
     1621 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
     1681 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
     1741 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
     1801 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
     1861 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
     1921 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
     1981 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
     2041 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
     2101 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
     2161 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
     2221 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
     2281 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
     2341 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
     2401 gcgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
     2461 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
     2521 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
     2581 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
     2641 ggcaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
     2701 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
     2761 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca
     2821 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg
     2881 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg
     2941 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca
     3001 ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg
     3061 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc
     3121 agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc
     3181 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac
     3241 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag
     3301 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc
     3361 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa
     3421 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac
     3481 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga
     3541 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat
     3601 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc
     3661 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg
     3721 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag
     3781 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc
     3841 acggttgaga cgtgcgtggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg
     3901 atctcttcgg taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt
     3961 cctcttggcg ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac
     4021 ggaaggcacc gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc
     4081 ggtacagggt cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt
     4141 cctggtcgat cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc
     4201 caaacttcac gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta
     4261 gggaacgctc gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg
     4321 tggtggtgtc ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc
     4381 gctcggcaat gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg
     4441 tcacaacgtc gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc
     4501 tggtgccggt gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc
     4561 gttggttggt caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg
     4621 cggcgctctt gttcatggcg taatgtctcc ggttctagtc gcaagtattc tactttatgc
     4681 gactaaaaca cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa
     4741 cttaggactt gtgcgacatg tcgttttcag aagacggctg cactgaacgt cagaagccga
     4801 ctgcactata gcagcggagg ggttggatca aagtactttg atcccgaggg gaaccctgtg
     4861 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
     4921 gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca
     4981 ctgatagttt aaactggcag acatactgtc ccacaaatga agatggaatc tgtaaaagaa
     5041 aacgcgtgaa ataatgcgtc tgacaaaggt taggtcggct gcctttaatc aataccaaag
     5101 tggtccctac cacgatggaa aaactgtgca gtcggtttgg ctttttctga cgaacaaata
     5161 agattcgtgg ccgacaggtg ggggtccacc atgtgaaggc atcttcagac tccaataatg
     5221 gagcaatgac gtaagggctt acgaaataag taagggtagt ttgggaaatg tccactcacc
     5281 cgtcagtcta taaatactta gcccctccct cattgttaag ggagcaaaat ctcagagaga
     5341 tagtcctaga gagagaaaga gagcaagtag cctagaagta gtcaaggcgg cgaagtattc
     5401 aggcacgtgg ccaggaagaa gaaaagccaa gacgacgaaa acaggtaaga gctaagcttg
     5461 ttcactgccg tataggcagc gagaccaagg tctcagttta agagctatgc tggaaacagc
     5521 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     5581 cggagaccaa ggtctcccgc ggttctatct agttacgcgt taaaccaact agaagttcac
     5641 tgccgtatag gcagggatcc tttctccata ataatgtgtg agtagttccc agataaggga
     5701 attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
     5761 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
     5821 actaaaatcc agatcggtac cgattagttt gtcaaatagt agagttcatt taaaattctt
     5881 cagccatata gttctatttt taagctagtc gacttttttt ttcttactga aaattaatat
     5941 ttttttcttt ttgaaatact aatacatcta aatttaacaa ttgccaaagt gatttttaat
     6001 tagcttgctg gctaatcaca ataaaaatta ctctccttta ctatataagt aaatttttat
     6061 tgctatattt gttattatta ttattattat taatatttat tttctacaaa tttaataata
     6121 ttttatttta tatcatttta aaaagataag taatgaaata ttaagaattc gtttataatt
     6181 cttttgcagg tgggtttcta tttgtaagct aatctttttc agttatcctt tttttaaaat
     6241 ctttattatt attatagcta tatcttttat cttttaaaat taacattatc tattaaagat
     6301 aatttcaata aaagagtaaa aattaattta gagttctact gtcttcaaat ttctatttta
     6361 aaaaatactt ttaaaacttg atgtattttt tacgtggttt ttcactatga cttaatttct
     6421 gttttattat aatatgtata aatataaaaa tagattttcc ataacatatt ataaaaaatg
     6481 taaggggcat ttacgtaaat agatagactt aaaagaggca ccgagtgaac cctaattctc
     6541 atcgttgaga ctataaaatg cccattatcc cattcgcaca gtctcttcat tacttttgct
     6601 gttatttctc ctcagctgtg ccgcatatcg cctaattttt cttctctaag gtttcatcat
     6661 cttcaccaat ttctttaatc tcgattcaat tttttatgtt tgatctgtta ttgttctgtc
     6721 actacatgtg tttttcagtt gttttactag atgattttca ctgtcttctt gttagatcat
     6781 acatatattg aaaatgtttt ggattgactt ttttgtattg tgaatatctg ttattgtttg
     6841 attgttgttc agtatttaca cacccgatct gtgttatgag cttggtcata actatttctc
     6901 tgtatgtaaa tacagatctg ttaatgtttg taatcaattt ttcatatgca ctgttgatat
     6961 tgttctctct cctgtcctgt tatatgttga tatgattcgg tttttgtata acttgaacta
     7021 aacactagtc ctaaatgttt tttttactat ttaagattta tataatatgg atagattttt
     7081 tgagttccta gtctctgaag aggttaagct tgctgtagtt gtttaccagt tgaggtgcaa
     7141 tactaaaaat caattcaatt actgatattt tttgctgttt aggtttttga caaagtactt
     7201 taatttgctt tattgaacta aaaacgtagt cctgaattca ttgcaagtgt gaaagctata
     7261 gttcattgtt tttgttgcaa ttcttgaaaa attaattggt caagctataa tggattttac
     7321 tttttctgtt ttaatattga atttgctgaa tttatgaatg ggttgcatgg tttttgaaat
     7381 atgttgttgt gtgttgtgta aatgcagttt cttagtgtct caacctagga tggaccacta
     7441 cctcgacatc aggctcaggc cagacccaga gttcccacca gcccagctca tgtccgtcct
     7501 cttcggcaag ctccaccagg ccctcgtggc ccagggcggc gacaggatcg gcgtgtcctt
     7561 cccagacctc gacgagtcca ggtccaggct cggcgagagg ctccgcatcc acgcctccgc
     7621 cgacgacctc agggccctcc tcgccaggcc gtggctggag ggcctcaggg accacctcca
     7681 gttcggcgag ccagccgtgg tgccacaccc aaccccatac aggcaagtgt ccagggtgca
     7741 agccaagtcc aacccagaga ggctcaggag gaggctcatg aggaggcacg acctctccga
     7801 ggaagaggcc aggaagcgca tcccagacac cgtggccagg gccctcgacc tcccattcgt
     7861 gaccctcagg tcccagtcca ccggccagca cttccgcctc ttcatcaggc acggcccact
     7921 ccaggtgacc gccgaggagg gcggctttac ctgctacggc ctctccaagg gcggcttcgt
     7981 gccgtggttc ggctccggcg ccaccaactt ctccctcctc aagcaagccg gcgacgtgga
     8041 ggagaaccca ggcccaatgc cagccgccaa gcgcgtcaag ctggacggcg gaaagcgcac
     8101 cgccgacgga tccgagttcg agagccctaa gaaaaagaga aaagtggaca agaagtactc
     8161 gatcggcctc gatattggga ctaactctgt tggctgggcc gtgatcaccg acgagtacaa
     8221 ggtgccctca aagaagttca aggtcctggg caacaccgat cggcattcca tcaagaagaa
     8281 tctcattggc gctctcctgt tcgacagcgg cgagacggct gaggctacgc ggctcaagcg
     8341 caccgcccgc aggcggtaca cgcgcaggaa gaatcgcatc tgctacctgc aggagatttt
     8401 ctccaacgag atggcgaagg ttgacgattc tttcttccac aggctggagg agtcattcct
     8461 cgtggaggag gataagaagc acgagcggca tccaatcttc ggcaacattg tcgacgaggt
     8521 tgcctaccac gagaagtacc ctacgatcta ccatctgcgg aagaagctcg tggactccac
     8581 agataaggcg gacctccgcc tgatctacct cgctctggcc cacatgatta agttcagggg
     8641 ccatttcctg atcgaggggg atctcaaccc ggacaatagc gatgttgaca agctgttcat
     8701 ccagctcgtg cagacgtaca accagctctt cgaggagaac cccattaatg cgtcaggcgt
     8761 cgacgcgaag gctatcctgt ccgctaggct ctcgaagtct cggaagctcg agaacctgat
     8821 cgcccagctg ccgggcgaga agaagaacgg cctgttcggg aatctcattg cgctcagcct
     8881 ggggctcacg cccaacttca agtcgaattt cgatctcgct gaggacgcca agctgcagct
     8941 ctccaaggac acatacgacg atgacctgga taacctcctg gcccagatcg gcgatcagta
     9001 cgcggacctg ttcctcgctg ccaagaatct gtcggacgcc atcctcctgt ctgatattct
     9061 cagggtgaac accgagatta cgaaggctcc gctctcagcc tccatgatca agcgctacga
     9121 cgagcaccat caggatctga ccctcctgaa ggcgctggtc aggcagcagc tccccgagaa
     9181 gtacaaggag atcttcttcg atcagtcgaa gaacggctac gctgggtaca ttgacggcgg
     9241 ggcctctcag gaggagttct acaagttcat caagccgatt ctggagaaga tggacggcac
     9301 ggaggagctg ctggtgaagc tcaagcgcga ggacctcctg aggaagcagc ggacattcga
     9361 taacggcagc atcccacacc agattcatct cggggagctg cacgctatcc tgaggaggca
     9421 ggaggacttc taccctttcc tcaaggataa ccgcgagaag atcgagaaga ttctgacttt
     9481 caggatcccg tactacgtcg gcccactcgc taggggcaac tcccgcttcg cttggatgac
     9541 ccgcaagtca gaggagacga tcacgccgtg gaacttcgag gaggtggtcg acaagggcgc
     9601 tagcgctcag tcgttcatcg agaggatgac gaatttcgac aagaacctgc caaatgagaa
     9661 ggtgctccct aagcactcgc tcctgtacga gtacttcaca gtctacaacg agctgactaa
     9721 ggtgaagtat gtgaccgagg gcatgaggaa gccggctttc ctgtctgggg agcagaagaa
     9781 ggccatcgtg gacctcctgt tcaagaccaa ccggaaggtc acggttaagc agctcaagga
     9841 ggactacttc aagaagattg agtgcttcga ttcggtcgag atctctggcg ttgaggaccg
     9901 cttcaacgcc tccctgggga cctaccacga tctcctgaag atcattaagg ataaggactt
     9961 cctggacaac gaggagaatg aggatatcct cgaggacatt gtgctgacac tcactctgtt
    10021 cgaggaccgg gagatgatcg aggagcgcct gaagacttac gcccatctct tcgatgacaa
    10081 ggtcatgaag cagctcaaga ggaggaggta caccggctgg gggaggctga gcaggaagct
    10141 catcaacggc attcgggaca agcagtccgg gaagacgatc ctcgacttcc tgaagagcga
    10201 tggcttcgcg aaccgcaatt tcatgcagct gattcacgat gacagcctca cattcaagga
    10261 ggatatccag aaggctcagg tgagcggcca gggggactcg ctgcacgagc atatcgcgaa
    10321 cctcgctggc tcgccagcta tcaagaaggg gattctgcag accgtgaagg ttgtggacga
    10381 gctggtgaag gtcatgggca ggcacaagcc tgagaacatc gtcattgaga tggcccggga
    10441 gaatcagacc acgcagaagg gccagaagaa ctcacgcgag aggatgaaga ggatcgagga
    10501 gggcattaag gagctggggt cccagatcct caaggagcac ccggtggaga acacgcagct
    10561 gcagaatgag aagctctacc tgtactacct ccagaatggc cgcgatatgt atgtggacca
    10621 ggagctggat attaacaggc tcagcgatta cgacgtcgat gccatcgttc cacagtcatt
    10681 cctgaaggat gactccattg acaacaaggt cctcaccagg tcggacaaga accggggcaa
    10741 gtctgataat gttccttcag aggaggtcgt taagaagatg aagaactact ggcgccagct
    10801 cctgaatgcc aagctgatca cgcagcggaa gttcgataac ctcacaaagg ctgagagggg
    10861 cgggctctct gagctggaca aggcgggctt catcaagagg cagctggtcg agacacggca
    10921 gatcactaag cacgttgcgc agattctcga ctcacggatg aacactaagt acgatgagaa
    10981 tgacaagctg atccgcgagg tgaaggtcat caccctgaag tcaaagctcg tctccgactt
    11041 caggaaggat ttccagttct acaaggttcg ggagatcaac aattaccacc atgcccatga
    11101 cgcgtacctg aacgcggtgg tcggcacagc tctgatcaag aagtacccaa agctcgagag
    11161 cgagttcgtg tacggggact acaaggttta cgatgtgagg aagatgatcg ccaagtcgga
    11221 gcaggagatt ggcaaggcta ccgccaagta cttcttctac tctaacatta tgaatttctt
    11281 caagacagag atcactctgg ccaatggcga gatccggaag cgccccctca tcgagacgaa
    11341 cggcgagacg ggggagatcg tgtgggacaa gggcagggat ttcgcgaccg tcaggaaggt
    11401 tctctccatg ccacaagtga atatcgtcaa gaagacagag gtccagactg gcgggttctc
    11461 taaggagtca attctgccta agcggaacag cgacaagctc atcgcccgca agaaggactg
    11521 ggatccgaag aagtacggcg ggttcgacag ccccactgtg gcctactcgg tcctggttgt
    11581 ggcgaaggtt gagaagggca agtccaagaa gctcaagagc gtgaaggagc tgctggggat
    11641 cacgattatg gagcgctcca gcttcgagaa gaacccgatc gatttcctgg aggcgaaggg
    11701 ctacaaggag gtgaagaagg acctgatcat taagctcccc aagtactcac tcttcgagct
    11761 ggagaacggc aggaagcgga tgctggcttc cgctggcgag ctgcagaagg ggaacgagct
    11821 ggctctgccg tccaagtatg tgaacttcct ctacctggcc tcccactacg agaagctcaa
    11881 gggcagcccc gaggacaacg agcagaagca gctgttcgtc gagcagcaca agcattacct
    11941 cgacgagatc attgagcaga tttccgagtt ctccaagcgc gtgatcctgg ccgacgcgaa
    12001 tctggataag gtcctctccg cgtacaacaa gcaccgcgac aagccaatca gggagcaggc
    12061 tgagaatatc attcatctct tcaccctgac gaacctcggc gcccctgctg ctttcaagta
    12121 cttcgacaca actatcgatc gcaagaggta cacaagcact aaggaggtcc tggacgcgac
    12181 cctcatccac cagtcgatta ccggcctcta cgagacgcgc atcgacctgt ctcagctcgg
    12241 gggcgacgaa ttctccggga gcgagacgcc aggcacctcc gagtcggcca ccccagaatc
    12301 tgccacagtg gtgtccggcc aaaagcagga ccgccagggc ggagaacgca gaaggtccca
    12361 gctcgatagg gatcagtgtg cctactgcaa ggagaagggc cactgggcca aagactgccc
    12421 gaaaaagccg cgcggcccac gcggcccaag gccacaaaca tccctccttc caaagaagaa
    12481 gcggaaggtg gagctcagcg gaggatcttc cggaggatct agcggctccg agacaccagg
    12541 aacatccgaa agcgctacac cagaatctag cggaggctct tccggaggat ctaggcctac
    12601 cctcaacatc gaggatgagt atcgcctcca cgaaacctcc aaagaaccgg acgtgtccct
    12661 cggcagcaca tggctcagcg acttcccaca agcgtgggcc gaaaccggcg gcatgggcct
    12721 cgccgtccgc caagccccac tcattatccc gctgaaggcg acctccacac cggtgtccat
    12781 caagcagtac ccgatgagcc aagaggcgag gctcgggatt aagccgcaca ttcagcgcct
    12841 cctcgatcaa ggcattctcg tgccgtgcca atccccgtgg aatacaccac tcctcccggt
    12901 caaaaagccg ggcaccaacg actatcgccc ggtccaagat ctccgcgagg tcaacaagcg
    12961 cgtggaagac atccacccga ccgtcccgaa cccgtataat ctgctctccg ggctcccacc
    13021 atcccaccag tggtatacag tgctggacct caaagacgcc ttcttctgtc tccgcctcca
    13081 cccaacaagc cagccgctct tcgccttcga gtggcgcgac ccggagatgg gcatctccgg
    13141 ccaactgaca tggacacgcc tcccgcaagg cttcaagaac agcccgacac tcttcaacga
    13201 ggcgctccat agggacctcg cggattttcg catccagcat ccggacctca tcctcctcca
    13261 gtatgccgat gatctcctcc tcgccgcgac ctccgagctg gattgtcaac aaggcacacg
    13321 cgcgctcctc caaacactcg ggaacctcgg ctatcgcgcg tccgcgaaaa aggcccaaat
    13381 ctgccagaag caagtgaagt acctcgggta tctgctcaag gaaggccaac gctggctcac
    13441 cgaagcgcgc aaagaaacag tgatggggca accgacaccg aaaacaccac gccagctgcg
    13501 cgagtttctc ggcaaagccg gcttctgtcg cctcttcatc ccgggctttg ccgagatggc
    13561 cgcgccactc tacccactca ccaagccggg cacactgttt aactgggggc cggatcagca
    13621 gaaagcctac caagagatca aacaagcgct cctcaccgcc ccagcgctcg ggctcccaga
    13681 tctcacaaag ccgttcgagc tgttcgtcga tgagaagcaa ggctacgcga agggcgtgct
    13741 cacacagaag ctcggcccgt ggaggaggcc agtggcctat ctctccaaaa aactcgatcc
    13801 agtggccgcc ggctggccac cgtgtctgcg catggtcgcc gcgattgccg tgctcacaaa
    13861 ggatgccggc aaactcacaa tgggccagcc gctggtgatc ctcgcgccac atgccgtgga
    13921 agccctcgtc aaacagccgc cggataggtg gctctccaat gcgcgcatga cccattacca
    13981 agcgctcctc ctcgacaccg atcgcgtcca gttcggccca gtggtcgccc tcaatccggc
    14041 gacactgctg ccactcccag aggagggcct ccaacacaac tgtctggata ttctcgcgga
    14101 agcgcatggc acaaggccag acctcacaga tcaaccgctc agcggcggca gcaaaaggac
    14161 cgccgattcc caacattcca ccccaccaaa aaccaagagg aaggtgggct ctggcccaaa
    14221 gaagaagagg aaggtctgag agctcagagc tttcgttcgt atcatcggtt tcgacaacgt
    14281 tcgtcaagtt caatgcatca gtttcattgc gcacacacca gaatcctact gagtttgagt
    14341 attatggcat tgggaaaact gtttttcttg taccatttgt tgtgcttgta atttactgtg
    14401 ttttttattc ggttttcgct atcgaactgt gaaatggaaa tggatggaga agagttaatg
    14461 aatgatatgg tccttttgtt cattctcaaa ttaatattat ttgttttttc tcttatttgt
    14521 tgtgtgttga atttgaaatt ataagagata tgcaaacatt ttgttttgag taaaaatgtg
    14581 tcaaatcgtg gcctctaatg accgaagtta atatgaggag taaaacactt gtagttgtac
    14641 cattatgctt attcactagg caacaaatat attttcagac ctagaaaagc tgcaaatgtt
    14701 actgaataca agtatgtcct cttgtgtttt agacatttat gaactttcct ttatgtaatt
    14761 ttccagaatc cttgtcagat tctaatcatt gctttataat tatagttata ctcatggatt
    14821 tgtagttgag tatgaaaata ttttttaatg cattttatga cttgccaatt gattgacaac
    14881 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    14941 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    15001 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    15061 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctagagcagc
    15121 ttgccaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
    15181 cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
    15241 gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag gaaggtggca
    15301 cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctgccgaca
    15361 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
    15421 ccacgtcttc aaagcaagtg gattgatgtg aacatggtgg agcacgacac tctcgtctac
    15481 tccaagaata tcaaagatac agtctcagaa gaccaaaggg ctattgagac ttttcaacaa
    15541 agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa
    15601 aggacagtag aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct
    15661 atcgttcaag atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
    15721 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc
    15781 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
    15841 taaggaagtt catttcattt ggagaggaca cgctgaaatc accagtctct ctctacaaat
    15901 ctatctctct cgagctttcg cagatccggg gggcaatgag atatgaaaaa gcctgaactc
    15961 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
    16021 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
    16081 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
    16141 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggagtt tagcgagagc
    16201 ctgacctatt gcatctcccg ccgttcacag ggtgtcacgt tgcaagacct gcctgaaacc
    16261 gaactgcccg ctgttctaca accggtcgcg gaggctatgg atgcgatcgc tgcggccgat
    16321 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
    16381 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
    16441 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
    16501 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
    16561 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
    16621 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
    16681 cgctacttcg agcggaggca tccggagctt gcaggatcgc cacgactccg ggcgtatatg
    16741 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
    16801 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
    16861 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
    16921 gatagtggaa accgacgccc cagcactcgt ccgagggcaa agaaatagag tagatgccga
    16981 ccgggatctg tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag
    17041 ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt
    17101 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
    17161 aatccagtac taaaatccag atcccccgaa ttaattcggc gttaattcag tacattaaaa
    17221 acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc
    17281 tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata
    17341 caggcagccc atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt
    17401 tgctcatgtt accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg
    17461 gatgatctcg cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga
    17521 tcaccgcggt ttcaaaatcg gctccgtcga tactatgtta tacgccaact ttgaaaacaa
    17581 ctttgaaaaa gctgttttct ggtatttaag gttttagaat gcaaggaaca gtgaattgga
    17641 gttcgtcttg ttataattag cttcttgggg tatctttaaa tactgtagaa aagaggaagg
    17701 aaataataaa tggctaaaat gagaatatca ccggaattga aaaaactgat cgaaaaatac
    17761 cgctgcgtaa aagatacgga aggaatgtct cctgctaagg tatataagct ggtgggagaa
    17821 aatgaaaacc tatatttaaa aatgacggac agccggtata aagggaccac ctatgatgtg
    17881 gaacgggaaa aggacatgat gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg
    17941 cactttgaac ggcatgatgg ctggagcaat ctgctcatga gtgaggccga tggcgtcctt
    18001 tgctcggaag agtatgaaga tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag
    18061 tgcatcaggc tctttcactc catcgacata tcggattgtc cctatacgaa tagcttagac
    18121 agccgcttag ccgaattgga ttacttactg aataacgatc tggccgatgt ggattgcgaa
    18181 aactgggaag aagacactcc atttaaagat ccgcgcgagc tgtatgattt tttaaagacg
    18241 gaaaagcccg aagaggaact tgtcttttcc cacggcgacc tgggagacag caacatcttt
    18301 gtgaaagatg gcaaagtaag tggctttatt gatcttggga gaagcggcag ggcggacaag
    18361 tggtatgaca ttgccttctg cgtccggtcg atcagggagg atatcgggga agaacagtat
    18421 gtcgagctat tttttgactt actggggatc aagcctgatt gggagaaaat aaaatattat
    18481 attttactgg atgaattgtt ttagtaccta gaatgcatga ccaaaatccc ttaacgtgag
    18541 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatctt
//
