LOCUS       sequence-454215-       15759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR/Cas-mediated gene editing in model fern species, 
            Ceratopteris richardii; expresses guide RNAs from CrActin promoter 
            and Cas9 from CaMV 35S promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15759)
  AUTHORS   Jiang W, Deng F, Babla M, Chen C, Yang D, Tong T, Qin Y, Chen G, 
            Marchant B, Soltis P, Soltis DE, Zeng F, Chen ZH
  TITLE     Efficient gene editing of a model fern species through 
            gametophyte-based transformation.
  JOURNAL   Plant Physiol. 2024 Sep 12:kiae473. doi: 10.1093/plphys/kiae473.
  PUBMED    39268871
REFERENCE   2  (bases 1 to 15759)
  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 
            Physiol. 2024 Sep 12:kiae473. doi: 10.1093/plphys/kiae473."
COMMENT     Sequence Label: pRGEB32-CrActin-35S
FEATURES             Location/Qualifiers
     source          1..15759
                     /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"
     primer_bind     5163..5185
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5177..5194
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5178..5194
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(5203..5455)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(5477..5500)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    5511..5535
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(5568..5615)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(5616..9716)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(5616..9716)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9741..9761)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9768..9833)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(9945..10621)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     terminator      complement(10733..10985)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12064..12080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12064..12080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12077..12099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12088..12104
                     /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(12112..12118,12119..12136,12137..12142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12157..12178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12369..13045
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             13112..14137
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    14178..14352
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    14430..14454
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             14879..15673
                     /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 cgtgctattc acaggaaaga gtgcttttcg 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 aacacatggg 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 aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca
     5041 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     5101 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     5161 ttcccagtca cgacgttgta aaacgacggc cagtgaattc ccgatctagt aacatagatg
     5221 acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt
     5281 aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt
     5341 acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa
     5401 gaccggcaac aggattcaat cttaagaaac tttattgcca aatgtttgaa cgatcgggga
     5461 aattcgagct ctaagccttg tcatcgtcat ccttgtagtc gctgttatca accactttgt
     5521 acaagaaagc tgggtctaga tatctcgagt gcggccgcga attcttactt tttctttttt
     5581 gcctggccgg cctttttcgt ggccgccggc cttttgtcgc ctcccagctg agacaggtcg
     5641 atccgtgtct cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc cagcacctct
     5701 ttggtgctgg tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa ggcggcaggg
     5761 gctcccagat tggtcagggt aaacaggtgg atgatattct cggcctgctc tctgatgggc
     5821 ttatcccggt gcttgttgta ggcggacagc actttgtcca gattagcgtc ggccaggatc
     5881 actctcttgg agaactcgct gatctgctcg atgatctcgt ccaggtagtg cttgtgctgt
     5941 tccacaaaca gctgtttctg ctcattatcc tcgggggagc ccttcagctt ctcatagtgg
     6001 ctggccaggt acaggaagtt cacatatttg gagggcaggg ccagttcgtt tcccttctgc
     6061 agttcgccgg cagaggccag cattctcttc cggccgtttt ccagctcgaa cagggagtac
     6121 ttaggcagct tgatgatcag gtcctttttc acttctttgt agcccttggc ttccagaaag
     6181 tcgatgggat tcttctcgaa gctgcttctt tccatgatgg tgatccccag cagctctttc
     6241 acactcttca gtttcttgga cttgcccttt tccactttgg ccaccaccag cacagaatag
     6301 gccacggtgg ggctgtcgaa gccgccgtac ttcttagggt cccagtcctt ctttctggcg
     6361 atcagcttat cgctgttcct cttgggcagg atagactctt tgctgaagcc gcctgtctgc
     6421 acctcggtct ttttcacgat attcacttgg ggcatgctca gcactttccg cacggtggca
     6481 aaatcccggc ccttatccca cacgatctcc ccggtttcgc cgtttgtctc gatcagaggc
     6541 cgcttccgga tctcgccgtt ggccagggta atctcggtct tgaaaaagtt catgatgttg
     6601 ctgtagaaga agtacttggc ggtagccttg ccgatttcct gctcgctctt ggcgatcatc
     6661 ttccgcacgt cgtacacctt gtagtcgccg tacacgaact cgctttccag cttagggtac
     6721 tttttgatca gggcggttcc cacgacggcg ttcaggtagg cgtcgtgggc gtggtggtag
     6781 ttgttgatct cgcgcacttt gtaaaactgg aaatccttcc ggaaatcgga caccagcttg
     6841 gacttcaggg tgatcacttt cacttcccgg atcagcttgt cattctcgtc gtacttagtg
     6901 ttcatccggg agtccaggat ctgtgccacg tgctttgtga tctgccgggt ttccaccagc
     6961 tgtctcttga tgaagccggc cttatccagt tcgctcaggc cgcctctctc ggccttggtc
     7021 agattgtcga actttctctg ggtaatcagc ttggcgttca gcagctgccg ccagtagttc
     7081 ttcatcttct tcacgacctc ttcggagggc acgttgtcgc tcttgccccg gttcttgtcg
     7141 cttctggtca gcaccttgtt gtcgatggag tcgtccttca gaaagctctg aggcacgata
     7201 tggtccacat cgtagtcgga cagccggttg atgtccagtt cctggtccac gtacatatcc
     7261 cgcccattct gcaggtagta caggtacagc ttctcgttct gcagctgggt gttttccacg
     7321 gggtgttctt tcaggatctg gctgcccagc tctttgatgc cctcttcgat ccgcttcatt
     7381 ctctcgcggc tgttcttctg tcccttctgg gtggtctggt tctctctggc catttcgatc
     7441 acgatgttct cgggcttgtg ccggcccatc actttcacga gctcgtccac caccttcact
     7501 gtctgcagga tgcccttctt aatggcgggg ctgccggcca gattggcaat gtgctcgtgc
     7561 aggctatcgc cctggccgga cacctgggct ttctggatgt cctctttaaa ggtcaggctg
     7621 tcgtcgtgga tcagctgcat gaagtttctg ttggcgaagc cgtcggactt caggaaatcc
     7681 aggattgtct tgccggactg cttgtcccgg atgccgttga tcagcttccg gctcagcctg
     7741 ccccagccgg tgtatctccg ccgcttcagc tgcttcatca ctttgtcgtc gaacaggtgg
     7801 gcataggttt tcagccgttc ctcgatcatc tctctgtcct caaacagtgt cagggtcagc
     7861 acgatatctt ccagaatgtc ctcgttttcc tcattgtcca ggaagtcctt gtccttgata
     7921 attttcagca gatcgtggta tgtgcccagg gaggcgttga accgatcttc cacgccggag
     7981 atttccacgg agtcgaagca ctcgattttc ttgaagtagt cctctttcag ctgcttcacg
     8041 gtcactttcc ggttggtctt gaacagcagg tccacgatgg cctttttctg ctcgccgctc
     8101 aggaaggcgg gctttctcat tccctcggtc acgtatttca ctttggtcag ctcgttatac
     8161 acggtgaagt actcgtacag caggctgtgc ttgggcagca ccttctcgtt gggcaggttc
     8221 ttatcgaagt tggtcatccg ctcgatgaag ctctgggcgg aagcgccctt gtccaccact
     8281 tcctcgaagt tccagggggt gatggtttcc tcgctctttc tggtcatcca ggcgaatctg
     8341 ctgtttcccc tggccagagg gcccacgtag taggggatgc ggaaggtcag gatcttctcg
     8401 atcttttccc ggttgtcctt caggaatggg taaaaatctt cctgccgccg cagaatggcg
     8461 tgcagctctc ccaggtggat ctggtggggg atgctgccgt tgtcgaaggt ccgctgcttc
     8521 cgcagcaggt cctctctgtt cagcttcacg agcagttcct cggtgccgtc catcttttcc
     8581 aggatgggct tgatgaactt gtagaactct tcctggctgg ctccgccgtc aatgtagccg
     8641 gcgtagccgt tcttgctctg gtcgaagaaa atctctttgt acttctcagg cagctgctgc
     8701 cgcacgagag ctttcagcag ggtcaggtcc tggtggtgct cgtcgtatct cttgatcata
     8761 gaggcgctca ggggggcctt ggtgatctcg gtgttcactc tcaggatgtc gctcagcagg
     8821 atggcgtcgg acaggttctt ggcggccaga aacaggtcgg cgtactggtc gccgatctgg
     8881 gccagcaggt tgtccaggtc gtcgtcgtag gtgtccttgc tcagctgcag tttggcatcc
     8941 tcggccaggt cgaagttgct cttgaagttg ggggtcaggc ccaggctcag ggcaatcagg
     9001 tttccgaaca ggccattctt cttctcgccg ggcagctggg cgatcagatt ttccagccgt
     9061 ctgctcttgc tcagtctggc agacaggatg gccttggcgt ccacgccgct ggcgttgatg
     9121 gggttttcct cgaacagctg gttgtaggtc tgcaccagct ggatgaacag cttgtccacg
     9181 tcgctgttgt cggggttcag gtcgccctcg atcaggaagt ggccccggaa cttgatcatg
     9241 tgggccaggg ccagatagat cagccgcagg tcggccttgt cggtgctgtc caccagtttc
     9301 tttctcaggt ggtagatggt ggggtacttc tcgtggtagg ccacctcgtc cacgatgttg
     9361 ccgaagatgg ggtgccgctc gtgcttctta tcctcttcca ccaggaagga ctcttccagt
     9421 ctgtggaaga agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag
     9481 cagatccggt tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc
     9541 tcggctgttt cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc
     9601 cggtcggtgt tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc
     9661 acggcccagc ccacagagtt ggtgccgatg tccaggccga tgctgtactt cttgtcggct
     9721 gctgggactc cgtggatacc gaccttccgc ttcttctttg gggccatctt atcgtcatcg
     9781 tctttgtaat caatatcatg atccttgtag tctccgtcgt ggtccttata gtccatggtt
     9841 aagtatttcc ttagagaatt ggttctatct ccttcgaaat ctcattgccc cccggatctg
     9901 cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg tgtcctctcc
     9961 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg
    10021 tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga
    10081 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
    10141 gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt gtaggtgcca
    10201 ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg gaatccgagg
    10261 aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact
    10321 gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt cacatcaatc
    10381 cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc tcgtgggtgg
    10441 gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct ttcctttatc
    10501 gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga tgaagtgaca
    10561 gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt gaaaagtctc
    10621 aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga cgagagtgtc
    10681 gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgcctct ccgatctagt
    10741 aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct
    10801 atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac
    10861 gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata
    10921 atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca aatgtttgaa
    10981 cgatccctgc aggcatgcac gcgctaaaaa cggactagcc ttattttaac ttgctatttc
    11041 tagctctaaa accgagacct cggtctccaa gagcatcagt taaatcccga cctgcaagat
    11101 ccagtcgcag gatagcatgg ggcaatgcat aaccctcata aatgggcact gtgtgcgtga
    11161 ctccatcacc agaatccagc acgatacctg agaagctcat caacccatga tttttttgga
    11221 cttagaaatt atatatgtat ttgttgaaat acaaatagca ttcaaggaaa gacataaaat
    11281 acatacctgt ggtccttcca cttgcataaa gtgataacac agcttgaatt gcaacataca
    11341 tagctggagc attgaacgtg tcaaacatga tctgtgtcat tttttctcga ttggccttcg
    11401 gattcaatgg agcttcagtg agcagaactg gatgctcctc tggagcaaca cgtagctcat
    11461 tatagaaagt atggtgccat atcttctcca tatcatccca attcgtcaca ataccatgct
    11521 cgatagggta cttcaatgta agaatcccac gctttgattg agcctcatca ccaacatatg
    11581 catctttttg gcccatacca accatgaccc cagtatgtct aggacgaccc acaatgctag
    11641 gaaatacagc ccgaggggca tcatcgccag caaatccagc ctgaattgaa gaaaatggat
    11701 gtgttgttgt tgtacatgaa gcaatagaca agaaaccaga caaaactttg gcattacctt
    11761 gaccatccca gaaccattat cacaaacaag cggttgaact tcgtccacat cagccatttt
    11821 cccttagctt tgttagataa tggacctagt tgataaaaca aatttaaaac ctgagttgaa
    11881 cgctaagcag cacgaaaata aaaaagatcc cttatacgaa acacagaaaa cacaatctaa
    11941 gctaaatagt gaaggtatgt aaaggcagaa ttaaagagat ctagaccata ttagcagatt
    12001 tctacacaat tttcatttat tttatagaat aagcagcaat atgagaaagc ttggcgtaat
    12061 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    12121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    12181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    12241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggc tagagcagct tgccaacatg
    12301 gtggagcacg acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccaa
    12361 agggctattg agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc
    12421 ccagctatct gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc
    12481 catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa
    12541 gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca
    12601 aagcaagtgg attgatgtga acatggtgga gcacgacact ctcgtctact ccaagaatat
    12661 caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa gggtaatatc
    12721 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa ggacagtaga
    12781 aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga
    12841 tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
    12901 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt
    12961 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
    13021 atttcatttg gagaggacac gctgaaatca ccagtctctc tctacaaatc tatctctctc
    13081 gagctttcgc agatccgggg ggcaatgaga tatgaaaaag cctgaactca ccgcgacgtc
    13141 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
    13201 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
    13261 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
    13321 cgcgctcccg attccggaag tgcttgacat tggggagttt agcgagagcc tgacctattg
    13381 catctcccgc cgttcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
    13441 tgttctacaa ccggtcgcgg aggctatgga tgcgatcgct gcggccgatc ttagccagac
    13501 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
    13561 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
    13621 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
    13681 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
    13741 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
    13801 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
    13861 gcggaggcat ccggagcttg caggatcgcc acgactccgg gcgtatatgc tccgcattgg
    13921 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
    13981 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
    14041 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
    14101 ccgacgcccc agcactcgtc cgagggcaaa gaaatagagt agatgccgac cgggatctgt
    14161 cgatcgacaa gctcgagttt ctccataata atgtgtgagt agttcccaga taagggaatt
    14221 agggttccta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt
    14281 gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtact
    14341 aaaatccaga tcccccgaat taattcggcg ttaattcagt acattaaaaa cgtccgcaat
    14401 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
    14461 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
    14521 tcagtccggg acggcgtcag cgggagagcc gttgtaaggc ggcagacttt gctcatgtta
    14581 ccgatgctat tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc
    14641 ggagggtagc atgttgattg taacgatgac agagcgttgc tgcctgtgat caccgcggtt
    14701 tcaaaatcgg ctccgtcgat actatgttat acgccaactt tgaaaacaac tttgaaaaag
    14761 ctgttttctg gtatttaagg ttttagaatg caaggaacag tgaattggag ttcgtcttgt
    14821 tataattagc ttcttggggt atctttaaat actgtagaaa agaggaagga aataataaat
    14881 ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa
    14941 agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct
    15001 atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa
    15061 ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg
    15121 gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga
    15181 gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct
    15241 ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc
    15301 cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga
    15361 agacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga
    15421 agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg
    15481 caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat
    15541 tgccttctgc gtccggtcga tcagggagga tatcggggaa gaacagtatg tcgagctatt
    15601 ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga
    15661 tgaattgttt tagtacctag aatgcatgac caaaatccct taacgtgagt tttcgttcca
    15721 ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
