LOCUS       sequence-454164-       17869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for SpCas9-D10A based A-to-G base editing for dicot 
            plants; NGG PAM; SpCas9D10A-ABE was driven by AtUBQ10 and the sgRNA 
            was driven by AtU3 promoter; Hygromycin for plants selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17869)
  AUTHORS   Cheng Y, Li G, Qi A, Mandlik R, Pan C, Wang D, Ge S, Qi Y
  TITLE     A comprehensive all-in-one CRISPR toolbox for large-scale screens in
            plants.
  JOURNAL   Plant Cell. 2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081.
  PUBMED    40202185
REFERENCE   2  (bases 1 to 17869)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2025 Apr 9:koaf081. doi: 10.1093/plcell/koaf081."
COMMENT     Sequence Label: pYPQAT31
FEATURES             Location/Qualifiers
     source          1..17869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1125..1319
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(1555..1577)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1663..1803
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1718..1737
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1818..1835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1989..2577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2069..2088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2664..3458)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    5082..5106
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    5184..5358
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(5399..6424)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(6491..7167)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(6534..6554)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    7358..7379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7394..7399,7400..7417,7418..7424)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7432..7448
                     /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)."
     primer_bind     7456..7472
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(7489..7741)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    7805..7829
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(8032..8107)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    8149..8170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8185..8190,8191..8208,8209..8215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8223..8239
                     /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)."
     primer_bind     8247..8263
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8259..8564
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGL"
     primer_bind     8282..8302
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8284..8302
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8349..8368)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8350..8368)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8378..8394)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8925..8945)
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(9620..9667)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(9668..13741,13742..13744,13745..13768))
                     /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             9747..9755
                     /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             complement(13793..13813)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(13820..13885)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(14486..14506)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(14546..14570)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(15253..15269)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    15472..15496
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             16797..17426
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(17855..17869,1..1059)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag
       61 gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg
      121 caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg
      181 tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat
      241 gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag
      301 cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt
      361 tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt
      421 tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc
      481 gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag
      541 cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag
      601 cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt
      661 agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta
      721 gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac
      781 cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc
      841 gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc
      901 gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg
      961 ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac
     1021 cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg
     1081 caaattgccc tagcagggga aaaaggtcga aaaggtctgt ttcctgtgga tagcacgtac
     1141 attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg
     1201 tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt
     1261 tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa
     1321 ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc
     1381 tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct
     1441 ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc
     1501 cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     1561 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     1621 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     1681 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     1741 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     1801 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     2041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2401 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     2461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2641 attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt
     2701 ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc
     2761 ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg
     2821 ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg
     2881 tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa
     2941 tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca
     3001 tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc
     3061 gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc
     3121 tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg
     3181 gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga
     3241 acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg
     3301 gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc
     3361 ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc
     3421 agttttttca attccggtga tattctcatt ttagccattt attatttcct tcctcttttc
     3481 tacagtattt aaagataccc caagaagcta aggaaggtgc gaataagcgg ggaaattctt
     3541 ctcggctgac tcagtcattt catttcttca tgtttgagcc gattttttct cccgtaaatg
     3601 ccttgaatca gcctatttag accgtttctt cgccatttaa ggcgttatcc ccagttttta
     3661 gtgagatctc tcccactgac gtatcatttg gtccgcccga aacaggttgg ccagcgtgaa
     3721 taacatcgcc agttggttat cgtttttcag caaccccttg tatctggctt tcacgaagcc
     3781 gaactgtcgc ttgatgatgc gaaatgggtg ctccaccctg gcccggatgc tggctttcat
     3841 gtattcgatg ttgatggccg ttttgttctt gcgtggatgc tgtttcaagg ttcttacctt
     3901 gccggggcgc tcggcgatca gccagtccac atccacctcg gccagctcct cgcgctgtgg
     3961 cgccccttgg tagccggcat cggctgagac aaattgctcc tctccatgca gcagattacc
     4021 cagctgattg aggtcatgct cgttggccgc ggtggtgacc aggctgtggg tcaggccact
     4081 cttggcatcg acaccaatgt gggccttcat gccaaagtgc cactgattgc ctttcttggt
     4141 ctgatgcatc tccggatcgc gttgctgctc tttgttcttg gtcgagctgg gtgcctcaat
     4201 gatggtggca tcgaccaagg tgccttgagt catcatgacg cctgcttcgg ccagccagcg
     4261 attgatggtc ttgaacaatt ggcgggccag ttgatgctgc tccagcaggt ggcggaaatt
     4321 catgatggtg gtgcggtccg gcaaggcgct atccagggat aaccgggcaa acagacgcat
     4381 ggaggcgatt tcgtacagag catcttccat cgcgccatcg ctcaggttgt accaatgctg
     4441 catgcagtga atgcgtagca tggtttccag cggataaggt cgccggccat taccagcctt
     4501 ggggtaaaac ggctcgatga cttccaccat gttttgccat ggcagaatct gctccatgcg
     4561 ggacaagaaa atctcttttc tggtctgacg gcgcttactg ctgaattcac tgtcggcgaa
     4621 ggtaagttga tgactcatga tgaaccctgt tctatggctc cagatgacaa acatgatctc
     4681 atatcaggga cttgttcgca ccttccctaa ttataacaag acgaactcca attcactgtt
     4741 ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt tttcaaagtt
     4801 ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac aggcagcaac
     4861 gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg tttcaaaccc
     4921 ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt ctgccgcctt
     4981 acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc gagtggtgat
     5041 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
     5101 gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactga
     5161 attaacgccg aattaattcg ggggatctgg attttagtac tggattttgg ttttaggaat
     5221 tagaaatttt attgatagaa gtattttaca aatacaaata catactaagg gtttcttata
     5281 tgctcaacac atgagcgaaa ccctatagga accctaattc ccttatctgg gaactactca
     5341 cacattatta tggagaaact cgagcttgtc gatcgacaga tcccggtcgg catctactct
     5401 atttctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta
     5461 cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc
     5521 cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat
     5581 tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga
     5641 gtcgtggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca
     5701 tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga
     5761 acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt
     5821 tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca
     5881 tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc
     5941 agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac
     6001 cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga
     6061 tcgcatccat agcctccgcg accggttgta gaacagcggg cagttcggtt tcaggcaggt
     6121 cttgcaacgt gacaccctgt gaacggcggg agatgcaata ggtcaggctc tcgctaaact
     6181 ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat
     6241 aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc
     6301 ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga
     6361 cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt
     6421 tcatatctca ttgccccccg gatctgcgaa agctcgagag agatagattt gtagagagag
     6481 actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt
     6541 cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat
     6601 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg
     6661 ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat
     6721 cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac
     6781 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct
     6841 caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt
     6901 cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct
     6961 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca
     7021 tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc accttccttt
     7081 tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
     7141 gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatctttg
     7201 atattcttgg agtagacgag agtgtcgtgc tccaccatgt tggcaagctg ctctagccaa
     7261 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7321 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7381 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7441 gataacaatt tcacacagga aacagctatg acatgattac gaattcccga tctagtaaca
     7501 tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg
     7561 cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca
     7621 tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca
     7681 tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat
     7741 cggggaaatt cgagctccac cgcggtggcg gccgctctag aactagttaa ttaagaatta
     7801 tcgaaccact ttgtacaaga aagctgggtc gaattcgccc ttactagttc acaatgtttt
     7861 aatttaggct cattatagat tctagttcca aaattcaaga attatctcta aatcagtcat
     7921 gaaagacatt cacatggatg ttggtttgtt ctaggttttc aatatctaca atgatcgata
     7981 tacaaattta agtttatggc aaaaaaaaga aaaaaggaaa gggaaaaaaa agcaccgact
     8041 cggtgccact ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct
     8101 ctaaaacaga gaccaccggt ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     8161 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     8221 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     8281 cgcaattaac cctcactaaa gggaacaaaa gctggaggac ctcgaggggg ggcccggtac
     8341 ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc
     8401 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     8461 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     8521 tgaatggcga atgggacgcg ccctgtagcg gcgccggtct ctgaccaatg gtgctccctc
     8581 agtgttatat atacaacaag atttgtttag atttgagcga tgtgggagaa attaggagct
     8641 tatatgaaac gcatacatgg gtactgggca tgggtattag tgaatatatg gacgatagcc
     8701 cattaaatta cataaaaagc ccacttatct tattgctcga tataagctcc gtatataaac
     8761 ggaagttgat cctaaaagtt cacttgtttc gtccaacata gccaaaacag atgttcttaa
     8821 acccagacgc catttgtttg attcagttat ccatcacagg ctgagccata agaaaaattt
     8881 aaagtaaata ggatcctgcg ttatctagag ggaacgccat ggggcaactt ttgtatacaa
     8941 agttgcgaat tcgccctttc cggtaagctt atctagagtt ccggaacaat tggcaagtca
     9001 taaaatgcat taaaaaatat tttcatactc aactacaaat ccatgagtat aactataatt
     9061 ataaagcaat gattagaatc tgacaaggat tctggaaaat tacataaagg aaagttcata
     9121 aatgtctaaa acacaagagg acatacttgt attcagtaac atttgcagct tttctaggtc
     9181 tgaaaatata tttgttgcct agtgaataag cataatggta caactacaag tgttttactc
     9241 ctcatattaa cttcggtcat tagaggccac gatttgacac atttttactc aaaacaaaat
     9301 gtttgcatat ctcttataat ttcaaattca acacacaaca aataagagaa aaaacaaata
     9361 atattaattt gagaatgaac aaaaggacca tatcattcat taactcttct ccatccattt
     9421 ccatttcaca gttcgatagc gaaaaccgaa taaaaaacac agtaaattac aagcacaaca
     9481 aatggtacaa gaaaaacagt tttcccaatg ccataatact caaactcagt aggattctgg
     9541 tgtgtgcgca atgaaactga tgcattgaac ttgacgaacg ttgtcgaaac cgatgatacg
     9601 aacgaaagct ctgagctcac ttcttcttct tcgcctgccc cgccttcttc gtcgccgctg
     9661 gccgcttgtc gcccccgagc tgagacaggt cgatgcgcgt ctcgtagagg ccggtaatcg
     9721 actggtggat gagggtcgcg tccaggacct ccttagtgct tgtgtacctc ttgcgatcga
     9781 tagttgtgtc gaagtacttg aaagcagcag gggcgccgag gttcgtcagg gtgaagagat
     9841 gaatgatatt ctcagcctgc tccctgattg gcttgtcgcg gtgcttgttg tacgcggaga
     9901 ggaccttatc cagattcgcg tcggccagga tcacgcgctt ggagaactcg gaaatctgct
     9961 caatgatctc gtcgaggtaa tgcttgtgct gctcgacgaa cagctgcttc tgctcgttgt
    10021 cctcggggct gcccttgagc ttctcgtagt gggaggccag gtagaggaag ttcacatact
    10081 tggacggcag agccagctcg ttccccttct gcagctcgcc agcggaagcc agcatccgct
    10141 tcctgccgtt ctccagctcg aagagtgagt acttggggag cttaatgatc aggtccttct
    10201 tcacctcctt gtagcccttc gcctccagga aatcgatcgg gttcttctcg aagctggagc
    10261 gctccataat cgtgatcccc agcagctcct tcacgctctt gagcttcttg gacttgccct
    10321 tctcaacctt cgccacaacc aggaccgagt aggccacagt ggggctgtcg aacccgccgt
    10381 acttcttcgg atcccagtcc ttcttgcggg cgatgagctt gtcgctgttc cgcttaggca
    10441 gaattgactc cttagagaac ccgccagtct ggacctctgt cttcttgacg atattcactt
    10501 gtggcatgga gagaaccttc ctgacggtcg cgaaatccct gcccttgtcc cacacgatct
    10561 cccccgtctc gccgttcgtc tcgatgaggg ggcgcttccg gatctcgcca ttggccagag
    10621 tgatctctgt cttgaagaaa ttcataatgt tagagtagaa gaagtacttg gcggtagcct
    10681 tgccaatctc ctgctccgac ttggcgatca tcttcctcac atcgtaaacc ttgtagtccc
    10741 cgtacacgaa ctcgctctcg agctttgggt acttcttgat cagagctgtg ccgaccaccg
    10801 cgttcaggta cgcgtcatgg gcatggtggt aattgttgat ctcccgaacc ttgtagaact
    10861 ggaaatcctt cctgaagtcg gagacgagct ttgacttcag ggtgatgacc ttcacctcgc
    10921 ggatcagctt gtcattctca tcgtacttag tgttcatccg tgagtcgaga atctgcgcaa
    10981 cgtgcttagt gatctgccgt gtctcgacca gctgcctctt gatgaagccc gccttgtcca
    11041 gctcagagag cccgcccctc tcagcctttg tgaggttatc gaacttccgc tgcgtgatca
    11101 gcttggcatt caggagctgg cgccagtagt tcttcatctt cttaacgacc tcctctgaag
    11161 gaacattatc agacttgccc cggttcttgt ccgacctggt gaggaccttg ttgtcaatgg
    11221 agtcatcctt caggaatgac tgtggaacga tatgatcgac gtcgtaatcg ctgagcctgt
    11281 taatatccag ctcctggtcc acatacatat cgcggccatt ctggaggtag tacaggtaga
    11341 gcttctcatt ctgcagctgc gtgttctcca ccgggtgctc cttgaggatc tgggacccca
    11401 gctccttaat gccctcctcg atcctcttca tcctctcgcg tgagttcttc tggcccttct
    11461 gcgtggtctg attctcccgg gccatctcaa tgacgatgtt ctcaggcttg tgcctgccca
    11521 tgaccttcac cagctcgtcc acaaccttca cggtctgcag aatccccttc ttgatagctg
    11581 gcgagccagc gaggttcgcg atatgctcgt gcagcgagtc cccctggccg ctcacctgag
    11641 ccttctggat atcctccttg aatgtgaggc tgtcatcgtg aatcagctgc atgaaattgc
    11701 ggttcgcgaa gccatcgctc ttcaggaagt cgaggatcgt cttcccggac tgcttgtccc
    11761 gaatgccgtt gatgagcttc ctgctcagcc tcccccagcc ggtgtacctc ctcctcttga
    11821 gctgcttcat gaccttgtca tcgaagagat gggcgtaagt cttcaggcgc tcctcgatca
    11881 tctcccggtc ctcgaacaga gtgagtgtca gcacaatgtc ctcgaggata tcctcattct
    11941 cctcgttgtc caggaagtcc ttatccttaa tgatcttcag gagatcgtgg taggtcccca
    12001 gggaggcgtt gaagcggtcc tcaacgccag agatctcgac cgaatcgaag cactcaatct
    12061 tcttgaagta gtcctccttg agctgcttaa ccgtgacctt ccggttggtc ttgaacagga
    12121 ggtccacgat ggccttcttc tgctccccag acaggaaagc cggcttcctc atgccctcgg
    12181 tcacatactt caccttagtc agctcgttgt agactgtgaa gtactcgtac aggagcgagt
    12241 gcttagggag caccttctca tttggcaggt tcttgtcgaa attcgtcatc ctctcgatga
    12301 acgactgagc gctagcgccc ttgtcgacca cctcctcgaa gttccacggc gtgatcgtct
    12361 cctctgactt gcgggtcatc caagcgaagc gggagttgcc cctagcgagt gggccgacgt
    12421 agtacgggat cctgaaagtc agaatcttct cgatcttctc gcggttatcc ttgaggaaag
    12481 ggtagaagtc ctcctgcctc ctcaggatag cgtgcagctc cccgagatga atctggtgtg
    12541 ggatgctgcc gttatcgaat gtccgctgct tcctcaggag gtcctcgcga ttgagcttca
    12601 ccagcagctc ctccgtgccg tccatcttct ccagaatcgg cttgatgaac ttgtagaact
    12661 cctcctgaga ggccccgccg tcaatgtacc cagcgtagcc gttcttcgac tgatcgaaga
    12721 agatctcctt gtacttctcg gggagctgct gcctgaccag cgccttcagg agggtcagat
    12781 cctgatggtg ctcgtcgtag cgcttgatca tggaggctga gagcggagcc ttcgtaatct
    12841 cggtgttcac cctgagaata tcagacagga ggatggcgtc cgacagattc ttggcagcga
    12901 ggaacaggtc cgcgtactga tcgccgatct gggccaggag gttatccagg tcatcgtcgt
    12961 atgtgtcctt ggagagctgc agcttggcgt cctcagcgag atcgaaattc gacttgaagt
    13021 tgggcgtgag ccccaggctg agcgcaatga gattcccgaa caggccgttc ttcttctcgc
    13081 ccggcagctg ggcgatcagg ttctcgaggc gccgagactt cgagagccta gcggacagga
    13141 tagccttcgc gtcgacgcct gacgcattaa tggggttctc ctcgaagagc tggttgtacg
    13201 tctgcacgag ctggatgaac agcttgtcaa catcgctatt gtccgggttg agatccccct
    13261 cgatcaggaa atggcccctg aacttaatca tgtgggccag agcgaggtag atcaggcgga
    13321 ggtccgcctt atctgtggag tccacgagct tcttccgcag atggtagatc gtagggtact
    13381 tctcgtggta ggcaacctcg tcgacaatgt tgccgaagat tggatgccgc tcgtgcttct
    13441 tatcctcctc cacgaggaat gactcctcca gcctgtggaa gaaagaatcg tcaaccttcg
    13501 ccatctcgtt ggagaaaatc tcctgcaggt agcagatgcg attcttcctg cgcgtgtacc
    13561 gcctgcgggc ggtgcgcttg agccgcgtag cctcagccgt ctcgccgctg tcgaacagga
    13621 gagcgccaat gagattcttc ttgatggaat gccgatcggt gttgcccagg accttgaact
    13681 tctttgaggg caccttgtac tcgtcggtga tcacggccca gccaacagag ttagtcccaa
    13741 tagcgaggcc gatcgagtac ttcttgtcag cagctggcac cccgtggatg ccaaccttcc
    13801 tcttcttctt cggagccatc ttgtcatcat catccttgta atcaatgtcg tggtccttgt
    13861 aatccccgtc gtggtccttg taatccatgg ttgacccccc gctgctgccc ccgctgcttt
    13921 caggtgttgc gctctcgctt gtgccaggtg tctcagagcc agaggagcct ccgctagatc
    13981 ctccggagtt gatggagctc tgggccttct tctgagcatt gaacacctgt ctaggcatcc
    14041 gatagaaatc gcacagcagg gcggcacatt catctgccag gattccctcg gtaatttcga
    14101 cgcggtgatt catgccgggg tagttcagca cgttcatcag ggagcctgcg gcgcctcttt
    14161 ttgagttcct cacgccaaac accacgcggc cgatcctaga gtggatcatg gcgccggcgc
    14221 acatcacgca aggctcgaat gtcacgtaca gggtggcgtc aatcagtctg tagttctgca
    14281 tgaccaggcc gccctgtctc agggccataa tttcggcatg ggctgttggg tcgtgcaggc
    14341 cgatggctct gttccagccc tcgccgatca ctctattgtt cagcaccagc acggctccca
    14401 caggcacctc cctctcatcc cgtgccctct tggccagggt cagggcatgt ctcatccagt
    14461 actcgtggga aaactccacc tcagagactt tccgcttctt ctttggtgac tcgaactcgc
    14521 ttccgtcggc tgtccgtttc atcggagcct gcttttttgt acaaacttgt ttgatagctt
    14581 ggcgcgcctc gagggggggc ccggtaccct gttaatcaga aaaactcaga ttaatctaca
    14641 aattcgatcg cacaaactag aaactaacac cagatctaga tagaaatcac aaatcgaaga
    14701 gtaattattc gacaaaactc aaattatttg aacaaatcgg atgatatcta tgaaacccta
    14761 atcgagaatt aagatgatat ctaacgatca aacccagaaa atcgtcttcg atctaagatt
    14821 aacagaatct aaaccaaaga acatatacga aattgggatc gaacgaaaac aaaatcgaag
    14881 attttgagag aataaggaac acagaaattt accttgatca cggtagagag aattgagaga
    14941 aagtttttaa gattttgaga aattgaaatc tgaattgtga agaagaagag ctctttgggt
    15001 attgttttat agaagaagaa gaagaaaaga cgaggacgac taggtcacga gaaagctaag
    15061 gcggtgaagc aatagctaat aataaaatga cacgtgtatt gagcgttgtt tacacgcaaa
    15121 gttgtttttg gctaattgcc ttatttttag gttgaggaaa agtatttgtg ctttgagttg
    15181 ataaacacga ctcgtgtgtg ccggctgcaa ccactttgac gccgtttatt actgactcgt
    15241 cgaaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
    15301 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
    15361 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgct agagcagctt
    15421 gagcttggat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt ttgacaggat
    15481 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
    15541 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc
    15601 tcgggatcaa agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg
    15661 ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc
    15721 tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa
    15781 tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct
    15841 gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca
    15901 cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc
    15961 ggagctggcc aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct
    16021 agaccgcctg gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc
    16081 cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg
    16141 catggtgttg accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg
    16201 cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac
    16261 cctcaccccg gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt
    16321 gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg
    16381 cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt
    16441 gaccgaggcc gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa
    16501 ccgcaccagg acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg
    16561 atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa
    16621 atcctggccg gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa
    16681 gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat
    16741 gcggtcgctg cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga
    16801 aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc
    16861 tagcccgcgc cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg
    16921 gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg
    16981 accgcccgac gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg
    17041 acggagcgcc ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc
    17101 tgattccggt gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg
    17161 ttaagcagcg cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg
    17221 cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc
    17281 ccattcttga gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca
    17341 caaccgttct tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg
    17401 ctgaaattaa atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac
    17461 aaacacgcta agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag
    17521 cctggcagac acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac
    17581 caagctgaag atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata
    17641 catcgcgcag ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg
    17701 ctaaaggagg cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca
    17761 tgtgtggagg aacgggcggt tggccaggcg taagcggctg ggttgcctgc cggccctgca
    17821 atggcactgg aacccccaag cccgaggaat cggcgtgagc ggtcgcaaa
//
