LOCUS       sequence-404911-       18602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cytosine base editing using miniSdd7 in monocotyledon plants 
            (Agrobacterium-mediated transformation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18602)
  AUTHORS   Huang J, Lin Q, Fei H, He Z, Xu H, Li Y, Qu K, Han P, Gao Q, Li B, 
            Liu G, Zhang L, Hu J, Zhang R, Zuo E, Luo Y, Ran Y, Qiu JL, Zhao KT,
            Gao C
  TITLE     Discovery of deaminase functions by structure-based protein 
            clustering.
  JOURNAL   Cell. 2023 Jul 20;186(15):3182-3195.e14. doi: 
            10.1016/j.cell.2023.05.041. Epub 2023 Jun 27.
  PUBMED    37379837
REFERENCE   2  (bases 1 to 18602)
  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: 
            "10.1016/j.cell.2023.05.041"; journalName: "Cell"; date: 
            "2023-07-20- 20"; volume: "186"; issue: "15"; pages: "3182-3195.e14"
COMMENT     Sequence Label: pH-Ubi-miniSdd7-nCas9-PBE
FEATURES             Location/Qualifiers
     source          1..18602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    21..45
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     233..255
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     247..264
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     248..264
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        331..710
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             990..1781
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        1933..2008
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2312..4304
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             4321..4341
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4867..4890,4891..4893,4894..8967)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             8880..8888
                     /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             8968..9015
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9037..9285
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9298..9318
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9977..9993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9977..9993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9990..10012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10001..10017
                     /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(10025..10031,10032..10049,10050..10055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10070..10091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10282..10958
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     10895..10915
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11025..12050
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    12091..12265
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    12343..12367
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             12792..13586
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      13673..14261
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14162..14181
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14415..14432
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    14447..14587
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(14513..14532)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14673..14695
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      14931..15125
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(15191..16264)
                     /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(16693..17322)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta
       61 aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc
      121 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      181 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      241 cgacgttgta aaacgacggc cagtgcctta taaagtaatt catccaggtc accaagttct
      301 aggattttca gaactgcaac ttattttatc aaggaatctt taaacatacg aacagatcac
      361 ttaaagttct tctgaagcaa cttaaagtta tcaggcatgc atggatcttg gaggaatcag
      421 atgtgcagtc agggaccata gcacaagaca ggcgtcttct actggtgcta ccagcaaatg
      481 ctggaagccg ggaacactgg gtacgttgga aaccacgtga tgtgaagaag taagataaac
      541 tgtaggagaa aagcatttcg tagtgggcca tgaagccttt caggacatgt attgcagtat
      601 gggccggccc attacgcaat tggacgacaa caaagactag tattagtacc acctcggcta
      661 tccacataga tcaaagctga tttaaaagag ttgtgcagat gatccgtggc gtgagaccaa
      721 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
      781 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
      841 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
      901 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
      961 gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga
     1021 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     1081 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     1141 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     1201 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     1261 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     1321 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     1381 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     1441 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     1501 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1561 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1621 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1681 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1741 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct agaaattcgt
     1801 tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac taagcacata
     1861 attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag cgtgcataat
     1921 aagccggtct cggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     1981 acttgaaaaa gtggcaccga gtcggtgctt ttttttttcg ttttgcattg agttttctcc
     2041 gtcgcatgtt tgcagtttta ttttccgttt tgcattgaaa tttctccgtc tcatgtttgc
     2101 agcgtgttca aaaagtacgc agctgtattt cacttattta cggcgccaca ttttcatgcc
     2161 gtttgtgcca actatcccga gctagtgaat acagcttggc ttcacacaac actggtgacc
     2221 cgctgacctg ctcgtacctc gtaccgtcgt acggcacagc atttggaatt aaagggtgtg
     2281 atcgatactg cttgctgcta agcttgcatg cctgcagtgc agcgtgaccc ggtcgtgccc
     2341 ctctctagag ataatgagca ttgcatgtct aagttataaa aaattaccac atattttttt
     2401 tgtcacactt gtttgaagtg cagtttatct atctttatac atatatttaa actttactct
     2461 acgaataata taatctatag tactacaata atatcagtgt tttagagaat catataaatg
     2521 aacagttaga catggtctaa aggacaattg agtattttga caacaggact ctacagtttt
     2581 atctttttag tgtgcatgtg ttctcctttt tttttgcaaa tagcttcacc tatataatac
     2641 ttcatccatt ttattagtac atccatttag ggtttagggt taatggtttt tatagactaa
     2701 tttttttagt acatctattt tattctattt tagcctctaa attaagaaaa ctaaaactct
     2761 attttagttt ttttatttaa taatttagat ataaaataga ataaaataaa gtgactaaaa
     2821 attaaacaaa taccctttaa gaaattaaaa aaactaagga aacatttttc ttgtttcgag
     2881 tagataatgc cagcctgtta aacgccgtcg acgagtctaa cggacaccaa ccagcgaacc
     2941 agcagcgtcg cgtcgggcca agcgaagcag acggcacggc atctctgtcg ctgcctctgg
     3001 acccctctcg agagttccgc tccaccgttg gacttgctcc gctgtcggca tccagaaatt
     3061 gcgtggcgga gcggcagacg tgagccggca cggcaggcgg cctcctcctc ctctcacggc
     3121 accggcagct acgggggatt cctttcccac cgctccttcg ctttcccttc ctcgcccgcc
     3181 gtaataaata gacaccccct ccacaccctc tttccccaac ctcgtgttgt tcggagcgca
     3241 cacacacaca accagatctc ccccaaatcc acccgtcggc acctccgctt caaggtacgc
     3301 cgctcgtcct cccccccccc ccctctctac cttctctaga tcggcgttcc ggtccatggt
     3361 tagggcccgg tagttctact tctgttcatg tttgtgttag atccgtgttt gtgttagatc
     3421 cgtgctgcta gcgttcgtac acggatgcga cctgtacgtc agacacgttc tgattgctaa
     3481 cttgccagtg tttctctttg gggaatcctg ggatggctct agccgttccg cagacgggat
     3541 cgatttcatg attttttttg tttcgttgca tagggtttgg tttgcccttt tcctttattt
     3601 caatatatgc cgtgcacttg tttgtcgggt catcttttca tgcttttttt tgtcttggtt
     3661 gtgatgatgt ggtctggttg ggcggtcgtt ctagatcgga gtagaattct gtttcaaact
     3721 acctggtgga tttattaatt ttggatctgt atgtgtgtgc catacatatt catagttacg
     3781 aattgaagat gatggatgga aatatcgatc taggataggt atacatgttg atgcgggttt
     3841 tactgatgca tatacagaga tgctttttgt tcgcttggtt gtgatgatgt ggtgtggttg
     3901 ggcggtcgtt cattcgttct agatcggagt agaatactgt ttcaaactac ctggtgtatt
     3961 tattaatttt ggaactgtat gtgtgtgtca tacatcttca tagttacgag tttaagatgg
     4021 atggaaatat cgatctagga taggtataca tgttgatgtg ggttttactg atgcatatac
     4081 atgatggcat atgcagcatc tattcatatg ctctaacctt gagtacctat ctattataat
     4141 aaacaagtat gttttataat tattttgatc ttgatatact tggatgatgg catatgcagc
     4201 agctatatgt ggattttttt agccctgcct tcatacgcta tttatttgct tggtactgtt
     4261 tcttttgtcg atgctcaccc tgttgtttgg tgttacttct gcagccctag ggccaccatg
     4321 ccaaagaaga agaggaaggt tgagggagga gggcctgggg cggttcctga aggaggcgac
     4381 ggcccgccag ccgtgccggc ggaggaggtg gagaggttga gaggagaact tcctccacct
     4441 gttgtcccgg gcactggtca aaaaacgcat gggcgctgga ttggtccaga tggtcgcgtc
     4501 cgcgccattg tttcaggaag ggatgaagat gctgcactgg tgcatgcaca gctcgccgcc
     4561 aaggggatac cagacgagcc aacaagaaat tctgatgttg agcagaagct ggccgcgcac
     4621 atggtggcaa atggcatccg ccacgtcacg ctcgtcatca accatagacc ctgccgcggc
     4681 ttcgacgaca gctgtgatac acttgtgccc ataattctgc cggagggctg caccctcacg
     4741 gtgcacgggc aaactgacaa gggcatgagg gtgcgggtaa ggtacaccgg cggcgcgcgg
     4801 ccgtggtggt ccaagtccgg cagcgagacg ccaggcacct ccgagagcgc tacgcctgaa
     4861 aggcctgaca agaagtactc gatcggcctc gccattggga ctaactctgt tggctgggcc
     4921 gtgatcaccg acgagtacaa ggtgccctca aagaagttca aggtcctggg caacaccgat
     4981 cggcattcca tcaagaagaa tctcattggc gctctcctgt tcgacagcgg cgagacggct
     5041 gaggctacgc ggctcaagcg caccgcccgc aggcggtaca cgcgcaggaa gaatcgcatc
     5101 tgctacctgc aggagatttt ctccaacgag atggcgaagg ttgacgattc tttcttccac
     5161 aggctggagg agtcattcct cgtggaggag gataagaagc acgagcggca tccaatcttc
     5221 ggcaacattg tcgacgaggt tgcctaccac gagaagtacc ctacgatcta ccatctgcgg
     5281 aagaagctcg tggactccac agataaggcg gacctccgcc tgatctacct cgctctggcc
     5341 cacatgatta agttcagggg ccatttcctg atcgaggggg atctcaaccc ggacaatagc
     5401 gatgttgaca agctgttcat ccagctcgtg cagacgtaca accagctctt cgaggagaac
     5461 cccattaatg cgtcaggcgt cgacgcgaag gctatcctgt ccgctaggct ctcgaagtct
     5521 cggcgcctcg agaacctgat cgcccagctg ccgggcgaga agaagaacgg cctgttcggg
     5581 aatctcattg cgctcagcct ggggctcacg cccaacttca agtcgaattt cgatctcgct
     5641 gaggacgcca agctgcagct ctccaaggac acatacgacg atgacctgga taacctcctg
     5701 gcccagatcg gcgatcagta cgcggacctg ttcctcgctg ccaagaatct gtcggacgcc
     5761 atcctcctgt ctgatattct cagggtgaac accgagatta cgaaggctcc gctctcagcc
     5821 tccatgatca agcgctacga cgagcaccat caggatctga ccctcctgaa ggcgctggtc
     5881 aggcagcagc tccccgagaa gtacaaggag atcttcttcg atcagtcgaa gaacggctac
     5941 gctgggtaca ttgacggcgg ggcctctcag gaggagttct acaagttcat caagccgatt
     6001 ctggagaaga tggacggcac ggaggagctg ctggtgaagc tcaatcgcga ggacctcctg
     6061 aggaagcagc ggacattcga taacggcagc atcccacacc agattcatct cggggagctg
     6121 cacgctatcc tgaggaggca ggaggacttc taccctttcc tcaaggataa ccgcgagaag
     6181 atcgagaaga ttctgacttt caggatcccg tactacgtcg gcccactcgc taggggcaac
     6241 tcccgcttcg cttggatgac ccgcaagtca gaggagacga tcacgccgtg gaacttcgag
     6301 gaggtggtcg acaagggcgc tagcgctcag tcgttcatcg agaggatgac gaatttcgac
     6361 aagaacctgc caaatgagaa ggtgctccct aagcactcgc tcctgtacga gtacttcaca
     6421 gtctacaacg agctgactaa ggtgaagtat gtgaccgagg gcatgaggaa gccggctttc
     6481 ctgtctgggg agcagaagaa ggccatcgtg gacctcctgt tcaagaccaa ccggaaggtc
     6541 acggttaagc agctcaagga ggactacttc aagaagattg agtgcttcga ttcggtcgag
     6601 atctctggcg ttgaggaccg cttcaacgcc tccctgggga cctaccacga tctcctgaag
     6661 atcattaagg ataaggactt cctggacaac gaggagaatg aggatatcct cgaggacatt
     6721 gtgctgacac tcactctgtt cgaggaccgg gagatgatcg aggagcgcct gaagacttac
     6781 gcccatctct tcgatgacaa ggtcatgaag cagctcaaga ggaggaggta caccggctgg
     6841 gggaggctga gcaggaagct catcaacggc attcgggaca agcagtccgg gaagacgatc
     6901 ctcgacttcc tgaagagcga tggcttcgcg aaccgcaatt tcatgcagct gattcacgat
     6961 gacagcctca cattcaagga ggatatccag aaggctcagg tgagcggcca gggggactcg
     7021 ctgcacgagc atatcgcgaa cctcgctggc tcgccagcta tcaagaaggg gattctgcag
     7081 accgtgaagg ttgtggacga gctggtgaag gtcatgggca ggcacaagcc tgagaacatc
     7141 gtcattgaga tggcccggga gaatcagacc acgcagaagg gccagaagaa ctcacgcgag
     7201 aggatgaaga ggatcgagga gggcattaag gagctggggt cccagatcct caaggagcac
     7261 ccggtggaga acacgcagct gcagaatgag aagctctacc tgtactacct ccagaatggc
     7321 cgcgatatgt atgtggacca ggagctggat attaacaggc tcagcgatta cgacgtcgat
     7381 catatcgttc cacagtcatt cctgaaggat gactccattg acaacaaggt cctcaccagg
     7441 tcggacaaga accggggcaa gtctgataat gttccttcag aggaggtcgt taagaagatg
     7501 aagaactact ggcgccagct cctgaatgcc aagctgatca cgcagcggaa gttcgataac
     7561 ctcacaaagg ctgagagggg cgggctctct gagctggaca aggcgggctt catcaagagg
     7621 cagctggtcg agacacggca gatcactaag cacgttgcgc agattctcga ctcacggatg
     7681 aacactaagt acgatgagaa tgacaagctg atccgcgagg tgaaggtcat caccctgaag
     7741 tcaaagctcg tctccgactt caggaaggat ttccagttct acaaggttcg ggagatcaac
     7801 aattaccacc atgcccatga cgcgtacctg aacgcggtgg tcggcacagc tctgatcaag
     7861 aagtacccaa agctcgagag cgagttcgtg tacggggact acaaggttta cgatgtgagg
     7921 aagatgatcg ccaagtcgga gcaggagatt ggcaaggcta ccgccaagta cttcttctac
     7981 tctaacatta tgaatttctt caagacagag atcactctgg ccaatggcga gatccggaag
     8041 cgccccctca tcgagacgaa cggcgagacg ggggagatcg tgtgggacaa gggcagggat
     8101 ttcgcgaccg tcaggaaggt tctctccatg ccacaagtga atatcgtcaa gaagacagag
     8161 gtccagactg gcgggttctc taaggagtca attctgccta agcggaacag cgacaagctc
     8221 atcgcccgca agaaggactg ggatccgaag aagtacggcg ggttcgacag ccccactgtg
     8281 gcctactcgg tcctggttgt ggcgaaggtt gagaagggca agtccaagaa gctcaagagc
     8341 gtgaaggagc tgctggggat cacgattatg gagcgctcca gcttcgagaa gaacccgatc
     8401 gatttcctgg aggcgaaggg ctacaaggag gtgaagaagg acctgatcat taagctcccc
     8461 aagtactcac tcttcgagct ggagaacggc aggaagcgga tgctggcttc cgctggcgag
     8521 ctgcagaagg ggaacgagct ggctctgccg tccaagtatg tgaacttcct ctacctggcc
     8581 tcccactacg agaagctcaa gggcagcccc gaggacaacg agcagaagca gctgttcgtc
     8641 gagcagcaca agcattacct cgacgagatc attgagcaga tttccgagtt ctccaagcgc
     8701 gtgatcctgg ccgacgcgaa tctggataag gtcctctccg cgtacaacaa gcaccgcgac
     8761 aagccaatca gggagcaggc tgagaatatc attcatctct tcaccctgac gaacctcggc
     8821 gcccctgctg ctttcaagta cttcgacaca actatcgatc gcaagaggta cacaagcact
     8881 aaggaggtcc tggacgcgac cctcatccac cagtcgatta ccggcctcta cgagacgcgc
     8941 atcgacctgt ctcagctcgg gggcgacaag cggccagcgg cgacgaagaa ggcggggcag
     9001 gcgaagaaga agaagacccg cgactccggc ggcagcacga acctctccga catcatcgag
     9061 aaggagacgg gcaagcagct cgtgatccag gagagcatcc tcatgctgcc ggaggaggtg
     9121 gaggaggtca tcggcaacaa gcccgagtcc gacatcctcg tgcacaccgc ctacgacgag
     9181 tccacggacg agaacgtcat gctcctgacg agcgacgctc cagagtacaa gccatgggct
     9241 ctcgtgatcc aggacagcaa cggcgagaac aagatcaaga tgctgtccgg cggctcccca
     9301 aagaagaagc ggaaggtgta ggagctcaga gctttcgttc gtatcatcgg tttcgacaac
     9361 gttcgtcaag ttcaatgcat cagtttcatt gcgcacacac cagaatccta ctgagtttga
     9421 gtattatggc attgggaaaa ctgtttttct tgtaccattt gttgtgcttg taatttactg
     9481 tgttttttat tcggttttcg ctatcgaact gtgaaatgga aatggatgga gaagagttaa
     9541 tgaatgatat ggtccttttg ttcattctca aattaatatt atttgttttt tctcttattt
     9601 gttgtgtgtt gaatttgaaa ttataagaga tatgcaaaca ttttgttttg agtaaaaatg
     9661 tgtcaaatcg tggcctctaa tgaccgaagt taatatgagg agtaaaacac ttgtagttgt
     9721 accattatgc ttattcacta ggcaacaaat atattttcag acctagaaaa gctgcaaatg
     9781 ttactgaata caagtatgtc ctcttgtgtt ttagacattt atgaactttc ctttatgtaa
     9841 ttttccagaa tccttgtcag attctaatca ttgctttata attatagtta tactcatgga
     9901 tttgtagttg agtatgaaaa tattttttaa tgcattttat gacttgccaa ttgattgaca
     9961 acgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    10021 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    10081 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    10141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggctagagca
    10201 gcttgccaac atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt
    10261 ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct
    10321 cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa aggaaggtgg
    10381 cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctgccga
    10441 cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc
    10501 aaccacgtct tcaaagcaag tggattgatg tgaacatggt ggagcacgac actctcgtct
    10561 actccaagaa tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac
    10621 aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca
    10681 aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg
    10741 ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
    10801 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
    10861 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta
    10921 tataaggaag ttcatttcat ttggagagga cacgctgaaa tcaccagtct ctctctacaa
    10981 atctatctct ctcgagcttt cgcagatccg gggggcaatg agatatgaaa aagcctgaac
    11041 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga
    11101 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat
    11161 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc
    11221 actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggag tttagcgaga
    11281 gcctgaccta ttgcatctcc cgccgttcac agggtgtcac gttgcaagac ctgcctgaaa
    11341 ccgaactgcc cgctgttcta caaccggtcg cggaggctat ggatgcgatc gctgcggccg
    11401 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta
    11461 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
    11521 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
    11581 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
    11641 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
    11701 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
    11761 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccacgactc cgggcgtata
    11821 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
    11881 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
    11941 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
    12001 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaagaaatag agtagatgcc
    12061 gaccgggatc tgtcgatcga caagctcgag tttctccata ataatgtgtg agtagttccc
    12121 agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc
    12181 ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc
    12241 aaaatccagt actaaaatcc agatcccccg aattaattcg gcgttaattc agtacattaa
    12301 aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat
    12361 cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga
    12421 tacaggcagc ccatcagtcc gggacggcgt cagcgggaga gccgttgtaa ggcggcagac
    12481 tttgctcatg ttaccgatgc tattcggaag aacggcaact aagctgccgg gtttgaaaca
    12541 cggatgatct cgcggagggt agcatgttga ttgtaacgat gacagagcgt tgctgcctgt
    12601 gatcaccgcg gtttcaaaat cggctccgtc gatactatgt tatacgccaa ctttgaaaac
    12661 aactttgaaa aagctgtttt ctggtattta aggttttaga atgcaaggaa cagtgaattg
    12721 gagttcgtct tgttataatt agcttcttgg ggtatcttta aatactgtag aaaagaggaa
    12781 ggaaataata aatggctaaa atgagaatat caccggaatt gaaaaaactg atcgaaaaat
    12841 accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa ggtatataag ctggtgggag
    12901 aaaatgaaaa cctatattta aaaatgacgg acagccggta taaagggacc acctatgatg
    12961 tggaacggga aaaggacatg atgctatggc tggaaggaaa gctgcctgtt ccaaaggtcc
    13021 tgcactttga acggcatgat ggctggagca atctgctcat gagtgaggcc gatggcgtcc
    13081 tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa gattatcgag ctgtatgcgg
    13141 agtgcatcag gctctttcac tccatcgaca tatcggattg tccctatacg aatagcttag
    13201 acagccgctt agccgaattg gattacttac tgaataacga tctggccgat gtggattgcg
    13261 aaaactggga agaagacact ccatttaaag atccgcgcga gctgtatgat tttttaaaga
    13321 cggaaaagcc cgaagaggaa cttgtctttt cccacggcga cctgggagac agcaacatct
    13381 ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg gagaagcggc agggcggaca
    13441 agtggtatga cattgccttc tgcgtccggt cgatcaggga ggatatcggg gaagaacagt
    13501 atgtcgagct attttttgac ttactgggga tcaagcctga ttgggagaaa ataaaatatt
    13561 atattttact ggatgaattg ttttagtacc tagaatgcat gaccaaaatc ccttaacgtg
    13621 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    13681 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    13741 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    13801 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
    13861 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    13921 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    13981 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    14041 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    14101 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    14161 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    14221 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    14281 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    14341 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
    14401 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
    14461 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct
    14521 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
    14581 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
    14641 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
    14701 accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg cggtcgagtg
    14761 gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccgtaggcca gccatttttg
    14821 agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag cgcagcgacc
    14881 gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt tatgcacagg
    14941 ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa aaatcgcctt
    15001 ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac ggctttgggt
    15061 tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg tacgtgctat
    15121 ccacaggaaa cagacctttt cgaccttttt cccctgctag ggcaatttgc cctagcatct
    15181 gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg tagcgcatga
    15241 ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc aggtcatttg
    15301 acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg ctgccactgc
    15361 gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca
    15421 ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca
    15481 gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct agctcgatct
    15541 cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta atcaaggctt
    15601 caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa
    15661 cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc tttccgccat
    15721 cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg cggccgggct
    15781 tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa accgccatca
    15841 gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa acctctacgt
    15901 gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc ttcgacagac
    15961 ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca tagagcatcg
    16021 gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac ggcgcaccgg
    16081 ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc cacgattcac
    16141 cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct
    16201 ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc
    16261 tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc aggcaaccca
    16321 gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca cggcgtcggt
    16381 gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc atctactcat
    16441 ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa
    16501 tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact
    16561 gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc
    16621 tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt
    16681 tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg
    16741 ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg
    16801 gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc
    16861 ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc
    16921 ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt
    16981 ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat
    17041 cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg
    17101 gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg
    17161 ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa
    17221 cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt
    17281 gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt
    17341 tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca
    17401 aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc
    17461 caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg cacggttgag
    17521 acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc gatctcttcg
    17581 gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt tcctcttggc
    17641 gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca cggaaggcac
    17701 cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg cggtacaggg
    17761 tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct tcctggtcga
    17821 tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg ccaaacttca
    17881 cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt agggaacgct
    17941 cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc gtggtggtgt
    18001 cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg cgctcggcaa
    18061 tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact gtcacaacgt
    18121 cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc ctggtgccgg
    18181 tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc cgttggttgg
    18241 tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg gcggcgctct
    18301 tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg cgactaaaac
    18361 acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta acttaggact
    18421 tgtgcgacat gtcgttttca gaagacggct gcactgaacg tcagaagccg actgcactat
    18481 agcagcggag gggttggatc aaagtacttt gatcccgagg ggaaccctgt ggttggcatg
    18541 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
    18601 aa
//
