LOCUS       sequence-454616-       15044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR/Cas-mediated gene editing in model fern species, 
            Ceratopteris richardii; expresses guide RNAs from CrU6-2 promoter 
            and Cas9 from CaMV 35S promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15044)
  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 15044)
  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-CrU6-2-35S
FEATURES             Location/Qualifiers
     source          1..15044
                     /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"
     primer_bind     complement(11349..11365)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11349..11365)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11362..11384)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11373..11389
                     /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(11397..11403,11404..11421,11422..11427))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11442..11463
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11654..12330
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             12397..13422
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    13463..13637
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    13715..13739
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             14164..14958
                     /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 cccctgcagg
    10741 catgcacgcg ctaaaaacgg actagcctta ttttaacttg ctatttctag ctctaaaacc
    10801 gagacctcgg tctccagcag ccaatcgctg gccggccaat atgtaggctc atctttcagg
    10861 gttgtggttc acatgtgggt tttctcctcc ccacattatt aataattcct atccacgcgc
    10921 gcctcttccg ttgaactacc ctcagtcgac acaccaacac actaaaatgc cacagctttt
    10981 caatcggcac aaaacccctc tgcaatacac ggacgaacaa aacagtgaga tcaaaggtag
    11041 tacgacaaac accatcacga agagggaaac gcaccgagaa cctcaaaaca tatggagcta
    11101 tgggagtcaa atgccccaga aacaagatta tatcttaaaa aatagctcca attccaattc
    11161 cacaaaccct cgaatcctag aaaaagctcc aattccaatt ctagaaagca tctttacaac
    11221 aaactacatt catagaaagt tcaacactga gcaccacccc ctcacgttct tccctcttta
    11281 ggagctcttt ttgtccgctc cgcacgtata tacatacacc tataacccta aaagcttggc
    11341 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    11401 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    11461 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    11521 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttggctagag cagcttgcca
    11581 acatggtgga gcacgacact ctcgtctact ccaagaatat caaagataca gtctcagaag
    11641 accaaagggc tattgagact tttcaacaaa gggtaatatc gggaaacctc ctcggattcc
    11701 attgcccagc tatctgtcac ttcatcaaaa ggacagtaga aaaggaaggt ggcacctaca
    11761 aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga tgcctctgcc gacagtggtc
    11821 ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt ccaaccacgt
    11881 cttcaaagca agtggattga tgtgaacatg gtggagcacg acactctcgt ctactccaag
    11941 aatatcaaag atacagtctc agaagaccaa agggctattg agacttttca acaaagggta
    12001 atatcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat caaaaggaca
    12061 gtagaaaagg aaggtggcac ctacaaatgc catcattgcg ataaaggaaa ggctatcgtt
    12121 caagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
    12181 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact
    12241 gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga
    12301 agttcatttc atttggagag gacacgctga aatcaccagt ctctctctac aaatctatct
    12361 ctctcgagct ttcgcagatc cggggggcaa tgagatatga aaaagcctga actcaccgcg
    12421 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
    12481 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
    12541 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
    12601 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg agtttagcga gagcctgacc
    12661 tattgcatct cccgccgttc acagggtgtc acgttgcaag acctgcctga aaccgaactg
    12721 cccgctgttc tacaaccggt cgcggaggct atggatgcga tcgctgcggc cgatcttagc
    12781 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
    12841 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
    12901 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
    12961 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
    13021 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
    13081 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
    13141 ttcgagcgga ggcatccgga gcttgcagga tcgccacgac tccgggcgta tatgctccgc
    13201 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
    13261 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
    13321 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
    13381 ggaaaccgac gccccagcac tcgtccgagg gcaaagaaat agagtagatg ccgaccggga
    13441 tctgtcgatc gacaagctcg agtttctcca taataatgtg tgagtagttc ccagataagg
    13501 gaattagggt tcctataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg
    13561 tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca
    13621 gtactaaaat ccagatcccc cgaattaatt cggcgttaat tcagtacatt aaaaacgtcc
    13681 gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac
    13741 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
    13801 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
    13861 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
    13921 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaccg
    13981 cggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga
    14041 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
    14101 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
    14161 taaatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc
    14221 gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa
    14281 aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg
    14341 gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt
    14401 gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg
    14461 gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc
    14521 aggctctttc actccatcga catatcggat tgtccctata cgaatagctt agacagccgc
    14581 ttagccgaat tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg
    14641 gaagaagaca ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag
    14701 cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa
    14761 gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat
    14821 gacattgcct tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag
    14881 ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta
    14941 ctggatgaat tgttttagta cctagaatgc atgaccaaaa tcccttaacg tgagttttcg
    15001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
