LOCUS       sequence-488643-       17684 bp    DNA     circular SYN 07-OCT-2025
DEFINITION  For plant prime editing including large DNA fragment editing in 
            Nicotiana benthamiana or other dicotyledons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17684)
  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 17684)
  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-2×35S-DualPE
FEATURES             Location/Qualifiers
     source          1..17684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..915
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(841..860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1001..1023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      1259..1453
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(1519..2592)
                     /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(3021..3650)
                     /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    4951..4975
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    5459..5478
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     protein_bind    5634..5653
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     polyA_signal    5660..5834
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     promoter        5841..6517
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             7094..7150
                     /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             7154..7180
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7220..7240
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             11254..11262
                     /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             11564..11584
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13310..13330
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(13989..14005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13989..14005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14002..14024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14013..14029
                     /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(14037..14043,14044..14061,14062..14067))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14082..14103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        14294..14970
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             15037..16062
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    16103..16277
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    16355..16379
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             16804..17598
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
      841 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
      901 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
      961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     1021 aggttttcac cgtcatcacc gaaacgcgcg aggcagggtg ccttgatgtg ggcgccggcg
     1081 gtcgagtggc gacggcgcgg cttgtccgcg ccctggtaga ttgcctggcc gtaggccagc
     1141 catttttgag cggccagcgg ccgcgatagg ccgacgcgaa gcggcggggc gtagggagcg
     1201 cagcgaccga agggtaggcg ctttttgcag ctcttcggct gtgcgctggc cagacagtta
     1261 tgcacaggcc aggcgggttt taagagtttt aataagtttt aaagagtttt aggcggaaaa
     1321 atcgcctttt ttctctttta tatcagtcac ttacatgtgt gaccggttcc caatgtacgg
     1381 ctttgggttc ccaatgtacg ggttccggtt cccaatgtac ggctttgggt tcccaatgta
     1441 cgtgctatcc acaggaaaca gaccttttcg acctttttcc cctgctaggg caatttgccc
     1501 tagcatctgc tccgtacatt aggaaccggc ggatgcttcg ccctcgatca ggttgcggta
     1561 gcgcatgact aggatcgggc cagcctgccc cgcctcctcc ttcaaatcgt actccggcag
     1621 gtcatttgac ccgatcagct tgcgcacggt gaaacagaac ttcttgaact ctccggcgct
     1681 gccactgcgt tcgtagatcg tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg
     1741 gcgtgccagg cggtagagaa aacggccgat gccgggatcg atcaaaaagt aatcggggtg
     1801 aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag
     1861 ctcgatctcg atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat
     1921 caaggcttca ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg
     1981 catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt
     2041 tccgccatcg gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg
     2101 gccgggcttg tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac
     2161 cgccatcagt accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac
     2221 ctctacgtgc ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt
     2281 cgacagacgg aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata
     2341 gagcatcgga acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacgg
     2401 cgcaccggct gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca
     2461 cgattcaccg gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt
     2521 caacttctcc accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt
     2581 gcgaccgctc acgccgattc ctcgggcttg ggggttccag tgccattgca gggccggcag
     2641 gcaacccagc cgcttacgcc tggccaaccg cccgttcctc cacacatggg gcattccacg
     2701 gcgtcggtgc ctggttgttc ttgattttcc atgccgcctc ctttagccgc taaaattcat
     2761 ctactcattt attcatttgc tcatttactc tggtagctgc gcgatgtatt cagatagcag
     2821 ctcggtaatg gtcttgcctt ggcgtaccgc gtacatcttc agcttggtgt gatcctccgc
     2881 cggcaactga aagttgaccc gcttcatggc tggcgtgtct gccaggctgg ccaacgttgc
     2941 agccttgctg ctgcgtgcgc tcggacggcc ggcacttagc gtgtttgtgc ttttgctcat
     3001 tttctcttta cctcattaac tcaaatgagt tttgatttaa tttcagcggc cagcgcctgg
     3061 acctcgcggg cagcgtcgcc ctcgggttct gattcaagaa cggttgtgcc ggcggcggca
     3121 gtgcctgggt agctcacgcg ctgcgtgata cgggactcaa gaatgggcag ctcgtacccg
     3181 gccagcgcct cggcaacctc accgccgatg cgcgtgcctt tgatcgcccg cgacacgaca
     3241 aaggccgctt gtagccttcc atccgtgacc tcaatgcgct gcttaaccag ctccaccagg
     3301 tcggcggtgg cccatatgtc gtaagggctt ggctgcaccg gaatcagcac gaagtcggct
     3361 gccttgatcg cggacacagc caagtccgcc gcctggggcg ctccgtcgat cactacgaag
     3421 tcgcgccggc cgatggcctt cacgtcgcgg tcaatcgtcg ggcggtcgat gccgacaacg
     3481 gttagcggtt gatcttcccg cacggccgcc caatcgcggg cactgccctg gggatcggaa
     3541 tcgactaaca gaacatcggc cccggcgagt tgcagggcgc gggctagatg ggttgcgatg
     3601 gtcgtcttgc ctgacccgcc tttctggtta agtacagcga taaccttcat gcgttcccct
     3661 tgcgtatttg tttatttact catcgcatca tatacgcagc gaccgcatga cgcaagctgt
     3721 tttactcaaa tacacatcac ctttttagac ggcggcgctc ggtttcttca gcggccaagc
     3781 tggccggcca ggccgccagc ttggcatcag acaaaccggc caggatttca tgcagccgca
     3841 cggttgagac gtgcgcgggc ggctcgaaca cgtacccggc cgcgatcatc tccgcctcga
     3901 tctcttcggt aatgaaaaac ggttcgtcct ggccgtcctg gtgcggtttc atgcttgttc
     3961 ctcttggcgt tcattctcgg cggccgccag ggcgtcggcc tcggtcaatg cgtcctcacg
     4021 gaaggcaccg cgccgcctgg cctcggtggg cgtcacttcc tcgctgcgct caagtgcgcg
     4081 gtacagggtc gagcgatgca cgccaagcag tgcagccgcc tctttcacgg tgcggccttc
     4141 ctggtcgatc agctcgcggg cgtgcgcgat ctgtgccggg gtgagggtag ggcgggggcc
     4201 aaacttcacg cctcgggcct tggcggcctc gcgcccgctc cgggtgcggt cgatgattag
     4261 ggaacgctcg aactcggcaa tgccggcgaa cacggtcaac accatgcggc cggccggcgt
     4321 ggtggtgtcg gcccacggct ctgccaggct acgcaggccc gcgccggcct cctggatgcg
     4381 ctcggcaatg tccagtaggt cgcgggtgct gcgggccagg cggtctagcc tggtcactgt
     4441 cacaacgtcg ccagggcgta ggtggtcaag catcctggcc agctccgggc ggtcgcgcct
     4501 ggtgccggtg atcttctcgg aaaacagctt ggtgcagccg gccgcgtgca gttcggcccg
     4561 ttggttggtc aagtcctggt cgtcggtgct gacgcgggca tagcccagca ggccagcggc
     4621 ggcgctcttg ttcatggcgt aatgtctccg gttctagtcg caagtattct actttatgcg
     4681 actaaaacac gcgacaagaa aacgccagga aaagggcagg gcggcagcct gtcgcgtaac
     4741 ttaggacttg tgcgacatgt cgttttcaga agacggctgc actgaacgtc agaagccgac
     4801 tgcactatag cagcggaggg gttggatcaa agtactttga tcccgagggg aaccctgtgg
     4861 ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt taaatatccg
     4921 attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac
     4981 tgatagttta aactggcaga catactgtcc cacaaatgaa gatggaatct gtaaaagaaa
     5041 acgcgtgaaa taatgcgtct gacaaaggtt aggtcggctg cctttaatca ataccaaagt
     5101 ggtccctacc acgatggaaa aactgtgcag tcggtttggc tttttctgac gaacaaataa
     5161 gattcgtggc cgacaggtgg gggtccacca tgtgaaggca tcttcagact ccaataatgg
     5221 agcaatgacg taagggctta cgaaataagt aagggtagtt tgggaaatgt ccactcaccc
     5281 gtcagtctat aaatacttag cccctccctc attgttaagg gagcaaaatc tcagagagat
     5341 agtcctagag agagaaagag agcaagtagc ctagaagtag tcaaggcggc gaagtattca
     5401 ggcacgtggc caggaagaag aaaagccaag acgacgaaaa caggtaagag ctaagcttgt
     5461 tcactgccgt ataggcagcg agaccaaggt ctcagtttaa gagctatgct ggaaacagca
     5521 tagcaagttt aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     5581 ggagaccaag gtctcccgcg gttctatcta gttacgcgtt aaaccaacta gaagttcact
     5641 gccgtatagg cagggatcct ttctccataa taatgtgtga gtagttccca gataagggaa
     5701 ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat
     5761 ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagta
     5821 ctaaaatcca gatcggtacc tgagactttt caacaaaggg taatatcggg aaacctcctc
     5881 ggattccatt gcccagctat ctgtcacttc atcaaaagga cagtagaaaa ggaaggtggc
     5941 acctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc ctctgccgac
     6001 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca
     6061 accacgtctt caaagcaagt ggattgatgt gaacatggtg gagcacgaca ctctcgtcta
     6121 ctccaagaat atcaaagata cagtctcaga agaccaaagg gctattgaga cttttcaaca
     6181 aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcaa
     6241 aaggacagta gaaaaggaag gtggcaccta caaatgccat cattgcgata aaggaaaggc
     6301 tatcgttcaa gatgcctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
     6361 catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt gatgtgatat
     6421 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
     6481 ataaggaagt tcatttcatt tggagaggac acgctgacct aggatggacc actacctcga
     6541 catcaggctc aggccagacc cagagttccc accagcccag ctcatgtccg tcctcttcgg
     6601 caagctccac caggccctcg tggcccaggg cggcgacagg atcggcgtgt ccttcccaga
     6661 cctcgacgag tccaggtcca ggctcggcga gaggctccgc atccacgcct ccgccgacga
     6721 cctcagggcc ctcctcgcca ggccgtggct ggagggcctc agggaccacc tccagttcgg
     6781 cgagccagcc gtggtgccac acccaacccc atacaggcaa gtgtccaggg tgcaagccaa
     6841 gtccaaccca gagaggctca ggaggaggct catgaggagg cacgacctct ccgaggaaga
     6901 ggccaggaag cgcatcccag acaccgtggc cagggccctc gacctcccat tcgtgaccct
     6961 caggtcccag tccaccggcc agcacttccg cctcttcatc aggcacggcc cactccaggt
     7021 gaccgccgag gagggcggct ttacctgcta cggcctctcc aagggcggct tcgtgccgtg
     7081 gttcggctcc ggcgccacca acttctccct cctcaagcaa gccggcgacg tggaggagaa
     7141 cccaggccca atgccagccg ccaagcgcgt caagctggac ggcggaaagc gcaccgccga
     7201 cggatccgag ttcgagagcc ctaagaaaaa gagaaaagtg gacaagaagt actcgatcgg
     7261 cctcgatatt gggactaact ctgttggctg ggccgtgatc accgacgagt acaaggtgcc
     7321 ctcaaagaag ttcaaggtcc tgggcaacac cgatcggcat tccatcaaga agaatctcat
     7381 tggcgctctc ctgttcgaca gcggcgagac ggctgaggct acgcggctca agcgcaccgc
     7441 ccgcaggcgg tacacgcgca ggaagaatcg catctgctac ctgcaggaga ttttctccaa
     7501 cgagatggcg aaggttgacg attctttctt ccacaggctg gaggagtcat tcctcgtgga
     7561 ggaggataag aagcacgagc ggcatccaat cttcggcaac attgtcgacg aggttgccta
     7621 ccacgagaag taccctacga tctaccatct gcggaagaag ctcgtggact ccacagataa
     7681 ggcggacctc cgcctgatct acctcgctct ggcccacatg attaagttca ggggccattt
     7741 cctgatcgag ggggatctca acccggacaa tagcgatgtt gacaagctgt tcatccagct
     7801 cgtgcagacg tacaaccagc tcttcgagga gaaccccatt aatgcgtcag gcgtcgacgc
     7861 gaaggctatc ctgtccgcta ggctctcgaa gtctcggaag ctcgagaacc tgatcgccca
     7921 gctgccgggc gagaagaaga acggcctgtt cgggaatctc attgcgctca gcctggggct
     7981 cacgcccaac ttcaagtcga atttcgatct cgctgaggac gccaagctgc agctctccaa
     8041 ggacacatac gacgatgacc tggataacct cctggcccag atcggcgatc agtacgcgga
     8101 cctgttcctc gctgccaaga atctgtcgga cgccatcctc ctgtctgata ttctcagggt
     8161 gaacaccgag attacgaagg ctccgctctc agcctccatg atcaagcgct acgacgagca
     8221 ccatcaggat ctgaccctcc tgaaggcgct ggtcaggcag cagctccccg agaagtacaa
     8281 ggagatcttc ttcgatcagt cgaagaacgg ctacgctggg tacattgacg gcggggcctc
     8341 tcaggaggag ttctacaagt tcatcaagcc gattctggag aagatggacg gcacggagga
     8401 gctgctggtg aagctcaagc gcgaggacct cctgaggaag cagcggacat tcgataacgg
     8461 cagcatccca caccagattc atctcgggga gctgcacgct atcctgagga ggcaggagga
     8521 cttctaccct ttcctcaagg ataaccgcga gaagatcgag aagattctga ctttcaggat
     8581 cccgtactac gtcggcccac tcgctagggg caactcccgc ttcgcttgga tgacccgcaa
     8641 gtcagaggag acgatcacgc cgtggaactt cgaggaggtg gtcgacaagg gcgctagcgc
     8701 tcagtcgttc atcgagagga tgacgaattt cgacaagaac ctgccaaatg agaaggtgct
     8761 ccctaagcac tcgctcctgt acgagtactt cacagtctac aacgagctga ctaaggtgaa
     8821 gtatgtgacc gagggcatga ggaagccggc tttcctgtct ggggagcaga agaaggccat
     8881 cgtggacctc ctgttcaaga ccaaccggaa ggtcacggtt aagcagctca aggaggacta
     8941 cttcaagaag attgagtgct tcgattcggt cgagatctct ggcgttgagg accgcttcaa
     9001 cgcctccctg gggacctacc acgatctcct gaagatcatt aaggataagg acttcctgga
     9061 caacgaggag aatgaggata tcctcgagga cattgtgctg acactcactc tgttcgagga
     9121 ccgggagatg atcgaggagc gcctgaagac ttacgcccat ctcttcgatg acaaggtcat
     9181 gaagcagctc aagaggagga ggtacaccgg ctgggggagg ctgagcagga agctcatcaa
     9241 cggcattcgg gacaagcagt ccgggaagac gatcctcgac ttcctgaaga gcgatggctt
     9301 cgcgaaccgc aatttcatgc agctgattca cgatgacagc ctcacattca aggaggatat
     9361 ccagaaggct caggtgagcg gccaggggga ctcgctgcac gagcatatcg cgaacctcgc
     9421 tggctcgcca gctatcaaga aggggattct gcagaccgtg aaggttgtgg acgagctggt
     9481 gaaggtcatg ggcaggcaca agcctgagaa catcgtcatt gagatggccc gggagaatca
     9541 gaccacgcag aagggccaga agaactcacg cgagaggatg aagaggatcg aggagggcat
     9601 taaggagctg gggtcccaga tcctcaagga gcacccggtg gagaacacgc agctgcagaa
     9661 tgagaagctc tacctgtact acctccagaa tggccgcgat atgtatgtgg accaggagct
     9721 ggatattaac aggctcagcg attacgacgt cgatgccatc gttccacagt cattcctgaa
     9781 ggatgactcc attgacaaca aggtcctcac caggtcggac aagaaccggg gcaagtctga
     9841 taatgttcct tcagaggagg tcgttaagaa gatgaagaac tactggcgcc agctcctgaa
     9901 tgccaagctg atcacgcagc ggaagttcga taacctcaca aaggctgaga ggggcgggct
     9961 ctctgagctg gacaaggcgg gcttcatcaa gaggcagctg gtcgagacac ggcagatcac
    10021 taagcacgtt gcgcagattc tcgactcacg gatgaacact aagtacgatg agaatgacaa
    10081 gctgatccgc gaggtgaagg tcatcaccct gaagtcaaag ctcgtctccg acttcaggaa
    10141 ggatttccag ttctacaagg ttcgggagat caacaattac caccatgccc atgacgcgta
    10201 cctgaacgcg gtggtcggca cagctctgat caagaagtac ccaaagctcg agagcgagtt
    10261 cgtgtacggg gactacaagg tttacgatgt gaggaagatg atcgccaagt cggagcagga
    10321 gattggcaag gctaccgcca agtacttctt ctactctaac attatgaatt tcttcaagac
    10381 agagatcact ctggccaatg gcgagatccg gaagcgcccc ctcatcgaga cgaacggcga
    10441 gacgggggag atcgtgtggg acaagggcag ggatttcgcg accgtcagga aggttctctc
    10501 catgccacaa gtgaatatcg tcaagaagac agaggtccag actggcgggt tctctaagga
    10561 gtcaattctg cctaagcgga acagcgacaa gctcatcgcc cgcaagaagg actgggatcc
    10621 gaagaagtac ggcgggttcg acagccccac tgtggcctac tcggtcctgg ttgtggcgaa
    10681 ggttgagaag ggcaagtcca agaagctcaa gagcgtgaag gagctgctgg ggatcacgat
    10741 tatggagcgc tccagcttcg agaagaaccc gatcgatttc ctggaggcga agggctacaa
    10801 ggaggtgaag aaggacctga tcattaagct ccccaagtac tcactcttcg agctggagaa
    10861 cggcaggaag cggatgctgg cttccgctgg cgagctgcag aaggggaacg agctggctct
    10921 gccgtccaag tatgtgaact tcctctacct ggcctcccac tacgagaagc tcaagggcag
    10981 ccccgaggac aacgagcaga agcagctgtt cgtcgagcag cacaagcatt acctcgacga
    11041 gatcattgag cagatttccg agttctccaa gcgcgtgatc ctggccgacg cgaatctgga
    11101 taaggtcctc tccgcgtaca acaagcaccg cgacaagcca atcagggagc aggctgagaa
    11161 tatcattcat ctcttcaccc tgacgaacct cggcgcccct gctgctttca agtacttcga
    11221 cacaactatc gatcgcaaga ggtacacaag cactaaggag gtcctggacg cgaccctcat
    11281 ccaccagtcg attaccggcc tctacgagac gcgcatcgac ctgtctcagc tcgggggcga
    11341 cgaattctcc gggagcgaga cgccaggcac ctccgagtcg gccaccccag aatctgccac
    11401 agtggtgtcc ggccaaaagc aggaccgcca gggcggagaa cgcagaaggt cccagctcga
    11461 tagggatcag tgtgcctact gcaaggagaa gggccactgg gccaaagact gcccgaaaaa
    11521 gccgcgcggc ccacgcggcc caaggccaca aacatccctc cttccaaaga agaagcggaa
    11581 ggtggagctc agcggaggat cttccggagg atctagcggc tccgagacac caggaacatc
    11641 cgaaagcgct acaccagaat ctagcggagg ctcttccgga ggatctaggc ctaccctcaa
    11701 catcgaggat gagtatcgcc tccacgaaac ctccaaagaa ccggacgtgt ccctcggcag
    11761 cacatggctc agcgacttcc cacaagcgtg ggccgaaacc ggcggcatgg gcctcgccgt
    11821 ccgccaagcc ccactcatta tcccgctgaa ggcgacctcc acaccggtgt ccatcaagca
    11881 gtacccgatg agccaagagg cgaggctcgg gattaagccg cacattcagc gcctcctcga
    11941 tcaaggcatt ctcgtgccgt gccaatcccc gtggaataca ccactcctcc cggtcaaaaa
    12001 gccgggcacc aacgactatc gcccggtcca agatctccgc gaggtcaaca agcgcgtgga
    12061 agacatccac ccgaccgtcc cgaacccgta taatctgctc tccgggctcc caccatccca
    12121 ccagtggtat acagtgctgg acctcaaaga cgccttcttc tgtctccgcc tccacccaac
    12181 aagccagccg ctcttcgcct tcgagtggcg cgacccggag atgggcatct ccggccaact
    12241 gacatggaca cgcctcccgc aaggcttcaa gaacagcccg acactcttca acgaggcgct
    12301 ccatagggac ctcgcggatt ttcgcatcca gcatccggac ctcatcctcc tccagtatgc
    12361 cgatgatctc ctcctcgccg cgacctccga gctggattgt caacaaggca cacgcgcgct
    12421 cctccaaaca ctcgggaacc tcggctatcg cgcgtccgcg aaaaaggccc aaatctgcca
    12481 gaagcaagtg aagtacctcg ggtatctgct caaggaaggc caacgctggc tcaccgaagc
    12541 gcgcaaagaa acagtgatgg ggcaaccgac accgaaaaca ccacgccagc tgcgcgagtt
    12601 tctcggcaaa gccggcttct gtcgcctctt catcccgggc tttgccgaga tggccgcgcc
    12661 actctaccca ctcaccaagc cgggcacact gtttaactgg gggccggatc agcagaaagc
    12721 ctaccaagag atcaaacaag cgctcctcac cgccccagcg ctcgggctcc cagatctcac
    12781 aaagccgttc gagctgttcg tcgatgagaa gcaaggctac gcgaagggcg tgctcacaca
    12841 gaagctcggc ccgtggagga ggccagtggc ctatctctcc aaaaaactcg atccagtggc
    12901 cgccggctgg ccaccgtgtc tgcgcatggt cgccgcgatt gccgtgctca caaaggatgc
    12961 cggcaaactc acaatgggcc agccgctggt gatcctcgcg ccacatgccg tggaagccct
    13021 cgtcaaacag ccgccggata ggtggctctc caatgcgcgc atgacccatt accaagcgct
    13081 cctcctcgac accgatcgcg tccagttcgg cccagtggtc gccctcaatc cggcgacact
    13141 gctgccactc ccagaggagg gcctccaaca caactgtctg gatattctcg cggaagcgca
    13201 tggcacaagg ccagacctca cagatcaacc gctcagcggc ggcagcaaaa ggaccgccga
    13261 ttcccaacat tccaccccac caaaaaccaa gaggaaggtg ggctctggcc caaagaagaa
    13321 gaggaaggtc tgagagctca gagctttcgt tcgtatcatc ggtttcgaca acgttcgtca
    13381 agttcaatgc atcagtttca ttgcgcacac accagaatcc tactgagttt gagtattatg
    13441 gcattgggaa aactgttttt cttgtaccat ttgttgtgct tgtaatttac tgtgtttttt
    13501 attcggtttt cgctatcgaa ctgtgaaatg gaaatggatg gagaagagtt aatgaatgat
    13561 atggtccttt tgttcattct caaattaata ttatttgttt tttctcttat ttgttgtgtg
    13621 ttgaatttga aattataaga gatatgcaaa cattttgttt tgagtaaaaa tgtgtcaaat
    13681 cgtggcctct aatgaccgaa gttaatatga ggagtaaaac acttgtagtt gtaccattat
    13741 gcttattcac taggcaacaa atatattttc agacctagaa aagctgcaaa tgttactgaa
    13801 tacaagtatg tcctcttgtg ttttagacat ttatgaactt tcctttatgt aattttccag
    13861 aatccttgtc agattctaat cattgcttta taattatagt tatactcatg gatttgtagt
    13921 tgagtatgaa aatatttttt aatgcatttt atgacttgcc aattgattga caacgaattc
    13981 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    14041 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    14101 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    14161 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggctagag cagcttgcca
    14221 acatggtgga gcacgacact ctcgtctact ccaagaatat caaagataca gtctcagaag
    14281 accaaagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc
    14341 attgcccagc tatctgtcac ttcatcaaaa ggacagtaga aaaggaaggt ggcacctaca
    14401 aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctgcc gacagtggtc
    14461 ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt
    14521 cttcaaagca agtggattga tgtgaacatg gtggagcacg acactctcgt ctactccaag
    14581 aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
    14641 atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca
    14701 gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt
    14761 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
    14821 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact
    14881 gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga
    14941 agttcatttc atttggagag gacacgctga aatcaccagt ctctctctac aaatctatct
    15001 ctctcgagct ttcgcagatc cggggggcaa tgagatatga aaaagcctga actcaccgcg
    15061 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
    15121 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
    15181 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
    15241 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg agtttagcga gagcctgacc
    15301 tattgcatct cccgccgttc acagggtgtc acgttgcaag acctgcctga aaccgaactg
    15361 cccgctgttc tacaaccggt cgcggaggct atggatgcga tcgctgcggc cgatcttagc
    15421 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
    15481 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
    15541 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
    15601 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
    15661 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
    15721 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
    15781 ttcgagcgga ggcatccgga gcttgcagga tcgccacgac tccgggcgta tatgctccgc
    15841 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
    15901 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
    15961 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
    16021 ggaaaccgac gccccagcac tcgtccgagg gcaaagaaat agagtagatg ccgaccggga
    16081 tctgtcgatc gacaagctcg agtttctcca taataatgtg tgagtagttc ccagataagg
    16141 gaattagggt tcctataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg
    16201 tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca
    16261 gtactaaaat ccagatcccc cgaattaatt cggcgttaat tcagtacatt aaaaacgtcc
    16321 gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac
    16381 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
    16441 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
    16501 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
    16561 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaccg
    16621 cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga
    16681 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
    16741 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
    16801 taaatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc
    16861 gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa
    16921 aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg
    16981 gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt
    17041 gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg
    17101 gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc
    17161 aggctctttc actccatcga catatcggat tgtccctata cgaatagctt agacagccgc
    17221 ttagccgaat tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg
    17281 gaagaagaca ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag
    17341 cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa
    17401 gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat
    17461 gacattgcct tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag
    17521 ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta
    17581 ctggatgaat tgttttagta cctagaatgc atgaccaaaa tcccttaacg tgagttttcg
    17641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
