LOCUS       sequence-488642-       19562 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For plant prime editing including large DNA fragment editing in 
            wheat plants or other monocotyledons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19562)
  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 19562)
  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/s41477-024-01898-3"; journalName: "Nat Plants"; date: 
            "2025-02"; volume: "11"; issue: "2"; pages: "191-205"
COMMENT     Sequence Label: pB–DualPE
FEATURES             Location/Qualifiers
     source          1..19562
                     /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"
     primer_bind     5164..5186
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5178..5195
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5179..5195
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5664..5683
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    5839..5858
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     polyA_signal    5865..6039
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        6046..8038
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             8624..8680
                     /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             8684..8710
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             8750..8770
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             12784..12792
                     /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             13094..13114
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14840..14860
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(15524..15540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15524..15540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15537..15559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15548..15564
                     /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(15572..15578,15579..15596,15597..15602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15617..15638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        15829..16174
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     16117..16137
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             16225..16776
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    16783..16957
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    17035..17059
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             18683..19477
                     /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 cgtgcgcggg 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 aaactgaagg cgggaaacga caatctgatc caagctcaag ctgctctagc
     5041 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5101 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     5161 tttcccagtc acgacgttgt aaaacgacgg ccagtgcctg gcagacatac tgtcccacaa
     5221 atgaagatgg aatctgtaaa agaaaacgcg tgaaataatg cgtctgacaa aggttaggtc
     5281 ggctgccttt aatcaatacc aaagtggtcc ctaccacgat ggaaaaactg tgcagtcggt
     5341 ttggcttttt ctgacgaaca aataagattc gtggccgaca ggtgggggtc caccatgtga
     5401 aggcatcttc agactccaat aatggagcaa tgacgtaagg gcttacgaaa taagtaaggg
     5461 tagtttggga aatgtccact cacccgtcag tctataaata cttagcccct ccctcattgt
     5521 taagggagca aaatctcaga gagatagtcc tagagagaga aagagagcaa gtagcctaga
     5581 agtagtcaag gcggcgaagt attcaggcac gtggccagga agaagaaaag ccaagacgac
     5641 gaaaacaggt aagagctaag cttgttcact gccgtatagg cagcgagacc aaggtctcag
     5701 tttaagagct atgctggaaa cagcatagca agtttaaata aggctagtcc gttatcaact
     5761 tgaaaaagtg gcaccgagtc ggtgcggaga ccaaggtctc ccgcggttct atctagttac
     5821 gcgttaaacc aactagaagt tcactgccgt ataggcaggg atcctttctc cataataatg
     5881 tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc atgtgttgag
     5941 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
     6001 taattcctaa aaccaaaatc cagtactaaa atccagatca agcttctgca gtgcagcgtg
     6061 acccggtcgt gcccctctct agagataatg agcattgcat gtctaagtta taaaaaatta
     6121 ccacatattt tttttgtcac acttgtttga agtgcagttt atctatcttt atacatatat
     6181 ttaaacttta ctctacgaat aatataatct atagtactac aataatatca gtgttttaga
     6241 gaatcatata aatgaacagt tagacatggt ctaaaggaca attgagtatt ttgacaacag
     6301 gactctacag ttttatcttt ttagtgtgca tgtgttctcc tttttttttg caaatagctt
     6361 cacctatata atacttcatc cattttatta gtacatccat ttagggttta gggttaatgg
     6421 tttttataga ctaatttttt tagtacatct attttattct attttagcct ctaaattaag
     6481 aaaactaaaa ctctatttta gtttttttat ttaataattt agatataaaa tagaataaaa
     6541 taaagtgact aaaaattaaa caaataccct ttaagaaatt aaaaaaacta aggaaacatt
     6601 tttcttgttt cgagtagata atgccagcct gttaaacgcc gtcgacgagt ctaacggaca
     6661 ccaaccagcg aaccagcagc gtcgcgtcgg gccaagcgaa gcagacggca cggcatctct
     6721 gtcgctgcct ctggacccct ctcgagagtt ccgctccacc gttggacttg ctccgctgtc
     6781 ggcatccaga aattgcgtgg cggagcggca gacgtgagcc ggcacggcag gcggcctcct
     6841 cctcctctca cggcaccggc agctacgggg gattcctttc ccaccgctcc ttcgctttcc
     6901 cttcctcgcc cgccgtaata aatagacacc ccctccacac cctctttccc caacctcgtg
     6961 ttgttcggag cgcacacaca cacaaccaga tctcccccaa atccacccgt cggcacctcc
     7021 gcttcaaggt acgccgctcg tcctcccccc ccccccctct ctaccttctc tagatcggcg
     7081 ttccggtcca tggttagggc ccggtagttc tacttctgtt catgtttgtg ttagatccgt
     7141 gtttgtgtta gatccgtgct gctagcgttc gtacacggat gcgacctgta cgtcagacac
     7201 gttctgattg ctaacttgcc agtgtttctc tttggggaat cctgggatgg ctctagccgt
     7261 tccgcagacg ggatcgattt catgattttt tttgtttcgt tgcatagggt ttggtttgcc
     7321 cttttccttt atttcaatat atgccgtgca cttgtttgtc gggtcatctt ttcatgcttt
     7381 tttttgtctt ggttgtgatg atgtggtctg gttgggcggt cgttctagat cggagtagaa
     7441 ttctgtttca aactacctgg tggatttatt aattttggat ctgtatgtgt gtgccataca
     7501 tattcatagt tacgaattga agatgatgga tggaaatatc gatctaggat aggtatacat
     7561 gttgatgcgg gttttactga tgcatataca gagatgcttt ttgttcgctt ggttgtgatg
     7621 atgtggtgtg gttgggcggt cgttcattcg ttctagatcg gagtagaata ctgtttcaaa
     7681 ctacctggtg tatttattaa ttttggaact gtatgtgtgt gtcatacatc ttcatagtta
     7741 cgagtttaag atggatggaa atatcgatct aggataggta tacatgttga tgtgggtttt
     7801 actgatgcat atacatgatg gcatatgcag catctattca tatgctctaa ccttgagtac
     7861 ctatctatta taataaacaa gtatgtttta taattatttt gatcttgata tacttggatg
     7921 atggcatatg cagcagctat atgtggattt ttttagccct gccttcatac gctatttatt
     7981 tgcttggtac tgtttctttt gtcgatgctc accctgttgt ttggtgttac ttctgcagcc
     8041 ctaggcttaa gtcatggacc actacctcga catcaggctc aggccagacc cagagttccc
     8101 accagcccag ctcatgtccg tcctcttcgg caagctccac caggccctcg tggcccaggg
     8161 cggcgacagg atcggcgtgt ccttcccaga cctcgacgag tccaggtcca ggctcggcga
     8221 gaggctccgc atccacgcct ccgccgacga cctcagggcc ctcctcgcca ggccgtggct
     8281 ggagggcctc agggaccacc tccagttcgg cgagccagcc gtggtgccac acccaacccc
     8341 atacaggcaa gtgtccaggg tgcaagccaa gtccaaccca gagaggctca ggaggaggct
     8401 catgaggagg cacgacctct ccgaggaaga ggccaggaag cgcatcccag acaccgtggc
     8461 cagggccctc gacctcccat tcgtgaccct caggtcccag tccaccggcc agcacttccg
     8521 cctcttcatc aggcacggcc cactccaggt gaccgccgag gagggcggct ttacctgcta
     8581 cggcctctcc aagggcggct tcgtgccgtg gttcggctcc ggcgccacca acttctccct
     8641 cctcaagcaa gccggcgacg tggaggagaa cccaggccca atgccagccg ccaagcgcgt
     8701 caagctggac ggcggaaagc gcaccgccga cggatccgag ttcgagagcc ctaagaaaaa
     8761 gagaaaagtg gacaagaagt actcgatcgg cctcgatatt gggactaact ctgttggctg
     8821 ggccgtgatc accgacgagt acaaggtgcc ctcaaagaag ttcaaggtcc tgggcaacac
     8881 cgatcggcat tccatcaaga agaatctcat tggcgctctc ctgttcgaca gcggcgagac
     8941 ggctgaggct acgcggctca agcgcaccgc ccgcaggcgg tacacgcgca ggaagaatcg
     9001 catctgctac ctgcaggaga ttttctccaa cgagatggcg aaggttgacg attctttctt
     9061 ccacaggctg gaggagtcat tcctcgtgga ggaggataag aagcacgagc ggcatccaat
     9121 cttcggcaac attgtcgacg aggttgccta ccacgagaag taccctacga tctaccatct
     9181 gcggaagaag ctcgtggact ccacagataa ggcggacctc cgcctgatct acctcgctct
     9241 ggcccacatg attaagttca ggggccattt cctgatcgag ggggatctca acccggacaa
     9301 tagcgatgtt gacaagctgt tcatccagct cgtgcagacg tacaaccagc tcttcgagga
     9361 gaaccccatt aatgcgtcag gcgtcgacgc gaaggctatc ctgtccgcta ggctctcgaa
     9421 gtctcggaag ctcgagaacc tgatcgccca gctgccgggc gagaagaaga acggcctgtt
     9481 cgggaatctc attgcgctca gcctggggct cacgcccaac ttcaagtcga atttcgatct
     9541 cgctgaggac gccaagctgc agctctccaa ggacacatac gacgatgacc tggataacct
     9601 cctggcccag atcggcgatc agtacgcgga cctgttcctc gctgccaaga atctgtcgga
     9661 cgccatcctc ctgtctgata ttctcagggt gaacaccgag attacgaagg ctccgctctc
     9721 agcctccatg atcaagcgct acgacgagca ccatcaggat ctgaccctcc tgaaggcgct
     9781 ggtcaggcag cagctccccg agaagtacaa ggagatcttc ttcgatcagt cgaagaacgg
     9841 ctacgctggg tacattgacg gcggggcctc tcaggaggag ttctacaagt tcatcaagcc
     9901 gattctggag aagatggacg gcacggagga gctgctggtg aagctcaagc gcgaggacct
     9961 cctgaggaag cagcggacat tcgataacgg cagcatccca caccagattc atctcgggga
    10021 gctgcacgct atcctgagga ggcaggagga cttctaccct ttcctcaagg ataaccgcga
    10081 gaagatcgag aagattctga ctttcaggat cccgtactac gtcggcccac tcgctagggg
    10141 caactcccgc ttcgcttgga tgacccgcaa gtcagaggag acgatcacgc cgtggaactt
    10201 cgaggaggtg gtcgacaagg gcgctagcgc tcagtcgttc atcgagagga tgacgaattt
    10261 cgacaagaac ctgccaaatg agaaggtgct ccctaagcac tcgctcctgt acgagtactt
    10321 cacagtctac aacgagctga ctaaggtgaa gtatgtgacc gagggcatga ggaagccggc
    10381 tttcctgtct ggggagcaga agaaggccat cgtggacctc ctgttcaaga ccaaccggaa
    10441 ggtcacggtt aagcagctca aggaggacta cttcaagaag attgagtgct tcgattcggt
    10501 cgagatctct ggcgttgagg accgcttcaa cgcctccctg gggacctacc acgatctcct
    10561 gaagatcatt aaggataagg acttcctgga caacgaggag aatgaggata tcctcgagga
    10621 cattgtgctg acactcactc tgttcgagga ccgggagatg atcgaggagc gcctgaagac
    10681 ttacgcccat ctcttcgatg acaaggtcat gaagcagctc aagaggagga ggtacaccgg
    10741 ctgggggagg ctgagcagga agctcatcaa cggcattcgg gacaagcagt ccgggaagac
    10801 gatcctcgac ttcctgaaga gcgatggctt cgcgaaccgc aatttcatgc agctgattca
    10861 cgatgacagc ctcacattca aggaggatat ccagaaggct caggtgagcg gccaggggga
    10921 ctcgctgcac gagcatatcg cgaacctcgc tggctcgcca gctatcaaga aggggattct
    10981 gcagaccgtg aaggttgtgg acgagctggt gaaggtcatg ggcaggcaca agcctgagaa
    11041 catcgtcatt gagatggccc gggagaatca gaccacgcag aagggccaga agaactcacg
    11101 cgagaggatg aagaggatcg aggagggcat taaggagctg gggtcccaga tcctcaagga
    11161 gcacccggtg gagaacacgc agctgcagaa tgagaagctc tacctgtact acctccagaa
    11221 tggccgcgat atgtatgtgg accaggagct ggatattaac aggctcagcg attacgacgt
    11281 cgatgccatc gttccacagt cattcctgaa ggatgactcc attgacaaca aggtcctcac
    11341 caggtcggac aagaaccggg gcaagtctga taatgttcct tcagaggagg tcgttaagaa
    11401 gatgaagaac tactggcgcc agctcctgaa tgccaagctg atcacgcagc ggaagttcga
    11461 taacctcaca aaggctgaga ggggcgggct ctctgagctg gacaaggcgg gcttcatcaa
    11521 gaggcagctg gtcgagacac ggcagatcac taagcacgtt gcgcagattc tcgactcacg
    11581 gatgaacact aagtacgatg agaatgacaa gctgatccgc gaggtgaagg tcatcaccct
    11641 gaagtcaaag ctcgtctccg acttcaggaa ggatttccag ttctacaagg ttcgggagat
    11701 caacaattac caccatgccc atgacgcgta cctgaacgcg gtggtcggca cagctctgat
    11761 caagaagtac ccaaagctcg agagcgagtt cgtgtacggg gactacaagg tttacgatgt
    11821 gaggaagatg atcgccaagt cggagcagga gattggcaag gctaccgcca agtacttctt
    11881 ctactctaac attatgaatt tcttcaagac agagatcact ctggccaatg gcgagatccg
    11941 gaagcgcccc ctcatcgaga cgaacggcga gacgggggag atcgtgtggg acaagggcag
    12001 ggatttcgcg accgtcagga aggttctctc catgccacaa gtgaatatcg tcaagaagac
    12061 agaggtccag actggcgggt tctctaagga gtcaattctg cctaagcgga acagcgacaa
    12121 gctcatcgcc cgcaagaagg actgggatcc gaagaagtac ggcgggttcg acagccccac
    12181 tgtggcctac tcggtcctgg ttgtggcgaa ggttgagaag ggcaagtcca agaagctcaa
    12241 gagcgtgaag gagctgctgg ggatcacgat tatggagcgc tccagcttcg agaagaaccc
    12301 gatcgatttc ctggaggcga agggctacaa ggaggtgaag aaggacctga tcattaagct
    12361 ccccaagtac tcactcttcg agctggagaa cggcaggaag cggatgctgg cttccgctgg
    12421 cgagctgcag aaggggaacg agctggctct gccgtccaag tatgtgaact tcctctacct
    12481 ggcctcccac tacgagaagc tcaagggcag ccccgaggac aacgagcaga agcagctgtt
    12541 cgtcgagcag cacaagcatt acctcgacga gatcattgag cagatttccg agttctccaa
    12601 gcgcgtgatc ctggccgacg cgaatctgga taaggtcctc tccgcgtaca acaagcaccg
    12661 cgacaagcca atcagggagc aggctgagaa tatcattcat ctcttcaccc tgacgaacct
    12721 cggcgcccct gctgctttca agtacttcga cacaactatc gatcgcaaga ggtacacaag
    12781 cactaaggag gtcctggacg cgaccctcat ccaccagtcg attaccggcc tctacgagac
    12841 gcgcatcgac ctgtctcagc tcgggggcga cgaattctcc gggagcgaga cgccaggcac
    12901 ctccgagtcg gccaccccag aatctgccac agtggtgtcc ggccaaaagc aggaccgcca
    12961 gggcggagaa cgcagaaggt cccagctcga tagggatcag tgtgcctact gcaaggagaa
    13021 gggccactgg gccaaagact gcccgaaaaa gccgcgcggc ccacgcggcc caaggccaca
    13081 aacatccctc cttccaaaga agaagcggaa ggtggagctc agcggaggat cttccggagg
    13141 atctagcggc tccgagacac caggaacatc cgaaagcgct acaccagaat ctagcggagg
    13201 ctcttccgga ggatctaggc ctaccctcaa catcgaggat gagtatcgcc tccacgaaac
    13261 ctccaaagaa ccggacgtgt ccctcggcag cacatggctc agcgacttcc cacaagcgtg
    13321 ggccgaaacc ggcggcatgg gcctcgccgt ccgccaagcc ccactcatta tcccgctgaa
    13381 ggcgacctcc acaccggtgt ccatcaagca gtacccgatg agccaagagg cgaggctcgg
    13441 gattaagccg cacattcagc gcctcctcga tcaaggcatt ctcgtgccgt gccaatcccc
    13501 gtggaataca ccactcctcc cggtcaaaaa gccgggcacc aacgactatc gcccggtcca
    13561 agatctccgc gaggtcaaca agcgcgtgga agacatccac ccgaccgtcc cgaacccgta
    13621 taatctgctc tccgggctcc caccatccca ccagtggtat acagtgctgg acctcaaaga
    13681 cgccttcttc tgtctccgcc tccacccaac aagccagccg ctcttcgcct tcgagtggcg
    13741 cgacccggag atgggcatct ccggccaact gacatggaca cgcctcccgc aaggcttcaa
    13801 gaacagcccg acactcttca acgaggcgct ccatagggac ctcgcggatt ttcgcatcca
    13861 gcatccggac ctcatcctcc tccagtatgc cgatgatctc ctcctcgccg cgacctccga
    13921 gctggattgt caacaaggca cacgcgcgct cctccaaaca ctcgggaacc tcggctatcg
    13981 cgcgtccgcg aaaaaggccc aaatctgcca gaagcaagtg aagtacctcg ggtatctgct
    14041 caaggaaggc caacgctggc tcaccgaagc gcgcaaagaa acagtgatgg ggcaaccgac
    14101 accgaaaaca ccacgccagc tgcgcgagtt tctcggcaaa gccggcttct gtcgcctctt
    14161 catcccgggc tttgccgaga tggccgcgcc actctaccca ctcaccaagc cgggcacact
    14221 gtttaactgg gggccggatc agcagaaagc ctaccaagag atcaaacaag cgctcctcac
    14281 cgccccagcg ctcgggctcc cagatctcac aaagccgttc gagctgttcg tcgatgagaa
    14341 gcaaggctac gcgaagggcg tgctcacaca gaagctcggc ccgtggagga ggccagtggc
    14401 ctatctctcc aaaaaactcg atccagtggc cgccggctgg ccaccgtgtc tgcgcatggt
    14461 cgccgcgatt gccgtgctca caaaggatgc cggcaaactc acaatgggcc agccgctggt
    14521 gatcctcgcg ccacatgccg tggaagccct cgtcaaacag ccgccggata ggtggctctc
    14581 caatgcgcgc atgacccatt accaagcgct cctcctcgac accgatcgcg tccagttcgg
    14641 cccagtggtc gccctcaatc cggcgacact gctgccactc ccagaggagg gcctccaaca
    14701 caactgtctg gatattctcg cggaagcgca tggcacaagg ccagacctca cagatcaacc
    14761 gctcagcggc ggcagcaaaa ggaccgccga ttcccaacat tccaccccac caaaaaccaa
    14821 gaggaaggtg ggctctggcc caaagaagaa gaggaaggtc tgaactagta gctcagagct
    14881 ttcgttcgta tcatcggttt cgacaacgtt cgtcaagttc aatgcatcag tttcattgcg
    14941 cacacaccag aatcctactg agtttgagta ttatggcatt gggaaaactg tttttcttgt
    15001 accatttgtt gtgcttgtaa tttactgtgt tttttattcg gttttcgcta tcgaactgtg
    15061 aaatggaaat ggatggagaa gagttaatga atgatatggt ccttttgttc attctcaaat
    15121 taatattatt tgttttttct cttatttgtt gtgtgttgaa tttgaaatta taagagatat
    15181 gcaaacattt tgttttgagt aaaaatgtgt caaatcgtgg cctctaatga ccgaagttaa
    15241 tatgaggagt aaaacacttg tagttgtacc attatgctta ttcactaggc aacaaatata
    15301 ttttcagacc tagaaaagct gcaaatgtta ctgaatacaa gtatgtcctc ttgtgtttta
    15361 gacatttatg aactttcctt tatgtaattt tccagaatcc ttgtcagatt ctaatcattg
    15421 ctttataatt atagttatac tcatggattt gtagttgagt atgaaaatat tttttaatgc
    15481 attttatgac ttgccaattg attgacaacg aattcgtaat catgtcatag ctgtttcctg
    15541 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
    15601 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
    15661 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
    15721 gaggcggttt gcgtattggc tagagcagct tgccaacatg gtggagcacg acactctcgt
    15781 ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca
    15841 acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
    15901 caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa
    15961 ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag
    16021 gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga
    16081 tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
    16141 tatataagga agttcatttc atttggagag gacacgctga aatcaccagt ctctctctac
    16201 aaatctatct ctctcgagtc taccatgagc ccagaacgac gcccggccga catccgccgt
    16261 gccaccgagg cggacatgcc ggcggtctgc accatcgtca accactacat cgagacaagc
    16321 acggtcaact tccgtaccga gccgcaggaa ccgcaggagt ggacggacga cctcgtccgt
    16381 ctgcgggagc gctatccctg gctcgtcgcc gaggtggacg gcgaggtcgc cggcatcgcc
    16441 tacgcgggcc cctggaaggc acgcaacgcc tacgactgga cggccgagtc gaccgtgtac
    16501 gtctcccccc gccaccagcg gacgggactg ggctccacgc tctacaccca cctgctgaag
    16561 tccctggagg cacagggctt caagagcgtg gtcgctgtca tcgggctgcc caacgacccg
    16621 agcgtgcgca tgcacgaggc gctcggatat gccccccgcg gcatgctgcg ggcggccggc
    16681 ttcaagcacg ggaactggca tgacgtgggt ttctggcagc tggacttcag cctgccggta
    16741 ccgccccgtc cggtcctgcc cgtcaccgag atttgactcg agtttctcca taataatgtg
    16801 tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca
    16861 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta
    16921 attcctaaaa ccaaaatcca gtactaaaat ccagatcccc cgaattaatt cggcgttaat
    16981 tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca atttgtttac
    17041 accacaatat atcctgccac cagccagcca acagctcccc gaccggcagc tcggcacaaa
    17101 atcaccactc gatacaggca gcccatcagt ccgggacggc gtcagcggga gagccgttgt
    17161 aaggcggcag actttgctca tgttaccgat gctattcgga agaacggcaa ctaagctgcc
    17221 gggtttgaaa cacggatgat ctcgcggagg gtagcatgtt gattgtaacg atgacagagc
    17281 gttgctgcct gtgatcaccg cggtttcaaa atcggctccg tcgatactat gttatacgcc
    17341 aactttgaaa acaactttga aaaagctgtt ttctggtatt taaggtttta gaatgcaagg
    17401 aacagtgaat tggagttcgt cttgttataa ttagggaagg tgcgaataag cggggaaatt
    17461 cttctcggct gactcagtca tttcatttct tcatgtttga gccgattttt tctcccgtaa
    17521 atgccttgaa tcagcctatt tagaccgttt cttcgccatt taaggcgtta tccccagttt
    17581 ttagtgagat ctctcccact gacgtatcat ttggtccgcc cgaaacaggt tggccagcgt
    17641 gaataacatc gccagttggt tatcgttttt cagcaacccc ttgtatctgg ctttcacgaa
    17701 gccgaactgt cgcttgatga tgcgaaatgg gtgctccacc ctggcccgga tgctggcttt
    17761 catgtattcg atgttgatgg ccgttttgtt cttgcgtgga tgctgtttca aggttcttac
    17821 cttgccgggg cgctcggcga tcagccagtc cacatccacc tcggccagct cctcgcgctg
    17881 tggcgcccct tggtagccgg catcggctga gacaaattgc tcctctccat gcagcagatt
    17941 acccagctga ttgaggtcat gctcgttggc cgcggtggtg accaggctgt gggtcaggcc
    18001 actcttggca tcgacaccaa tgtgggcctt catgccaaag tgccactgat tgcctttctt
    18061 ggtctgatgc atctccggat cgcgttgctg ctctttgttc ttggtcgagc tgggtgcctc
    18121 aatgatggtg gcatcgacca aggtgccttg agtcatcatg acgcctgctt cggccagcca
    18181 gcgattgatg gtcttgaaca attggcgggc cagttgatgc tgctccagca ggtggcggaa
    18241 attcatgatg gtggtgcggt ccggcaaggc gctatccagg gataaccggg caaacagacg
    18301 catggaggcg atttcgtaca gagcatcttc catcgcgcca tcgctcaggt tgtaccaatg
    18361 ctgcatgcag tgaatgcgta gcatggtttc cagcggataa ggtcgccggc cattaccagc
    18421 cttggggtaa aacggctcga tgacttccac catgttttgc catggcagaa tctgctccat
    18481 gcgggacaag aaaatctctt ttctggtctg acggcgctta ctgctgaatt cactgtcggc
    18541 gaaggtaagt tgatgactca tgatgaaccc tgttctatgg ctccagatga caaacatgat
    18601 ctcatatcag ggacttgttc gcaccttcct tagcttcttg gggtatcttt aaatactgta
    18661 gaaaagagga aggaaataat aaatggctaa aatgagaata tcaccggaat tgaaaaaact
    18721 gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg tctcctgcta aggtatataa
    18781 gctggtggga gaaaatgaaa acctatattt aaaaatgacg gacagccggt ataaagggac
    18841 cacctatgat gtggaacggg aaaaggacat gatgctatgg ctggaaggaa agctgcctgt
    18901 tccaaaggtc ctgcactttg aacggcatga tggctggagc aatctgctca tgagtgaggc
    18961 cgatggcgtc ctttgctcgg aagagtatga agatgaacaa agccctgaaa agattatcga
    19021 gctgtatgcg gagtgcatca ggctctttca ctccatcgac atatcggatt gtccctatac
    19081 gaatagctta gacagccgct tagccgaatt ggattactta ctgaataacg atctggccga
    19141 tgtggattgc gaaaactggg aagaagacac tccatttaaa gatccgcgcg agctgtatga
    19201 ttttttaaag acggaaaagc ccgaagagga acttgtcttt tcccacggcg acctgggaga
    19261 cagcaacatc tttgtgaaag atggcaaagt aagtggcttt attgatcttg ggagaagcgg
    19321 cagggcggac aagtggtatg acattgcctt ctgcgtccgg tcgatcaggg aggatatcgg
    19381 ggaagaacag tatgtcgagc tattttttga cttactgggg atcaagcctg attgggagaa
    19441 aataaaatat tatattttac tggatgaatt gttttagtac ctagaatgca tgaccaaaat
    19501 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    19561 tt
//
