LOCUS       sequence-358594-       16308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 in dividing Arabidopsis cells. Contains dsRED and 
            Basta for fast transformant selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16308)
  AUTHORS   Bieluszewski T, Sura W, Dziegielewski W, Bieluszewska A, Lachance C,
            Kabza M, Szymanska-Lejman M, Abram M, Wlodzimierz P, De Winne N, De 
            Jaeger G, Sadowski J, Cote J, Ziolkowski PA
  TITLE     NuA4 and H2A.Z control environmental responses and autotrophic 
            growth in Arabidopsis.
  JOURNAL   Nat Commun. 2022 Jan 12;13(1):277. doi: 10.1038/s41467-021-27882-5.
  PUBMED    35022409
REFERENCE   2  (bases 1 to 16308)
  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: "Nat 
            Commun."; date: "2022-01-12"; volume: "13(1):277. doi"; pages: " 
            10.1038/s41467-021-27882-5"
COMMENT     Sequence Label: pFGC-I2Cas9
FEATURES             Location/Qualifiers
     source          1..16308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1762..1786
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     1974..1996
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1988..2005
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1989..2005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3832..3897
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3904..3924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3949..8049
                     /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             3949..8049
                     /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             8050..8097
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8136..8388
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     terminator      10234..10486
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        10505..10885
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             10891..11442
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      11452..11704
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    11790..11814
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             12239..13033
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      13120..13708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13609..13628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13862..13879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    13894..14034
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(13960..13979)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14120..14142
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      14378..14572
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(14638..15711)
                     /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(join(16140..16308,1..461))
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct
       61 tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg
      121 gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc
      181 gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg
      241 ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt
      301 tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac
      361 agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg
      421 cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt
      481 gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa
      541 atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc
      601 aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggttgaga
      661 cgtgcgcggg cggctcgaac acgtacccgg ccgcgatcat ctccgcctcg atctcttcgg
      721 taatgaaaaa cggttcgtcc tggccgtcct ggtgcggttt catgcttgtt cctcttggcg
      781 ttcattctcg gcggccgcca gggcgtcggc ctcggtcaat gcgtcctcac ggaaggcacc
      841 gcgccgcctg gcctcggtgg gcgtcacttc ctcgctgcgc tcaagtgcgc ggtacagggt
      901 cgagcgatgc acgccaagca gtgcagccgc ctctttcacg gtgcggcctt cctggtcgat
      961 cagctcgcgg gcgtgcgcga tctgtgccgg ggtgagggta gggcgggggc caaacttcac
     1021 gcctcgggcc ttggcggcct cgcgcccgct ccgggtgcgg tcgatgatta gggaacgctc
     1081 gaactcggca atgccggcga acacggtcaa caccatgcgg ccggccggcg tggtggtgtc
     1141 ggcccacggc tctgccaggc tacgcaggcc cgcgccggcc tcctggatgc gctcggcaat
     1201 gtccagtagg tcgcgggtgc tgcgggccag gcggtctagc ctggtcactg tcacaacgtc
     1261 gccagggcgt aggtggtcaa gcatcctggc cagctccggg cggtcgcgcc tggtgccggt
     1321 gatcttctcg gaaaacagct tggtgcagcc ggccgcgtgc agttcggccc gttggttggt
     1381 caagtcctgg tcgtcggtgc tgacgcgggc atagcccagc aggccagcgg cggcgctctt
     1441 gttcatggcg taatgtctcc ggttctagtc gcaagtattc tactttatgc gactaaaaca
     1501 cgcgacaaga aaacgccagg aaaagggcag ggcggcagcc tgtcgcgtaa cttaggactt
     1561 gtgcgacatg tcgttttcag aagacggctg cactgaacgt cagaagccga ctgcactata
     1621 gcagcggagg ggttggatca aagtactttg atcccgaggg gaaccctgtg gttggcatgc
     1681 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta
     1741 ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt
     1801 aaactgaagg cgggaaacga caatctgatc caagctcaag ctgctctagc attcgccatt
     1861 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     1921 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     1981 acgacgttgt aaaacgacgg ccagtgccaa gctcaacgct gccagattcg atgtaagtta
     2041 ctggaaactg aaccgaaggg aactctccag gaaacagaga tggaaaactg aaactaactc
     2101 tgtttccaaa cgttagggtt ccttgaaatt gcctaccaat tgttcccgct ccaaccgtca
     2161 taatccacgg cgctccgtta atcaaactcc agtaatatgg cccgtcgtta ccgccggaag
     2221 taaccacgaa aacgcctttt tgaatcgctg caaaagatgc gactgcgatt ggatcatttt
     2281 ctagaccgaa accatcattg tcgtcatcat cctcaaaact caatcctagc gacagagata
     2341 tcacatggac tccatctcga atcgcttgat caatcgctgc gattacatct gatgagtaaa
     2401 tcccttcttc ccacgcggct ttgtagattg ctagatgcgc gtgtggtgcg attcctgagg
     2461 cggttccttg ggcgtaagag aagtaagacg cgttcttaac atgattccca gctgcaatgg
     2521 ctgcaacatg agtgccgtgt ccgattgtat cgtacggtga agagtattgg ccaatcttgg
     2581 tttctctcaa atcagggttg ttggcgaaca aacccttgtt gaatacctta gcaccgatta
     2641 gtttcttgtt gcacaaggaa gaagaattga attcacatgc tcctttccat ttagacggaa
     2701 ccgaaccaac cccatcgtcg tgaaagctgg gactatcggg ccaaattccg gtatctataa
     2761 taccgattac tataccagct ccataattcg agaccggcca tgtaccggat gtcgaattta
     2821 aaccgataaa ctttggagag aaagtggtat gaagcttaac cggtaaatct ttggtgaacg
     2881 agacataccc gggtttgtgt ttaagacgtt ggagctcaga gttggtgaga accgcgctaa
     2941 acccgtgaac cgaatcggta taagcgtaga taatctttgg ttttctattg gttataactg
     3001 aagtaagagt tgtggaaaac cagctccgat gatcggaaaa tggcaacggt ttagcggata
     3061 agtccatgtg gatgatgtaa ggagaggtct ccgccgttac aatcgctacg aaaaatgaga
     3121 acaccagaat aattacgacg gtcatcccca tgtctgagca agtttttttc tctggtctct
     3181 tcaaagtgaa atgaattttg gtgttagtaa gaaacaatta tagtgttgtt gttaatagat
     3241 gtttagtttt tattatgaca aaaaagttgc agagaatgta acctttactt ggcttttgct
     3301 tttgttaaag tggttttagg tggtcggcaa cacatcatct ctcatcattg tttttttttc
     3361 ttttctcttt caattgggct ttatcataga gaacaatgat tttggatatg aacttattag
     3421 tagtttcagc aatagtttag actttcgaat cggtaaatat cagtaactta gaactcaggc
     3481 tgaaactttg attcttttac tgtatttagc tagattctac acttggtgga ctccatcagc
     3541 aattacttca cagaccgagg tagctcctac aactctctga tgcttgatct cgttcaaata
     3601 gatgtctcat aattgcttct tttttgtttt tgtttttggg ctttattaaa gaggagtaag
     3661 gcccattaag agaacccgtt tgtttgttgg attcggattt ggcaaatgga tagcgccaaa
     3721 atcgtgtcga ttttcccgcc atctccattt cttaatctgg gttcttaaac ctaaatcagt
     3781 tactcagctt caccaaacca ccaaattgac cggatttgta aaatcgagat ggactataag
     3841 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
     3901 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
     3961 agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     4021 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     4081 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
     4141 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     4201 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     4261 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     4321 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     4381 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     4441 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     4501 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     4561 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     4621 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     4681 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     4741 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     4801 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     4861 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     4921 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     4981 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     5041 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     5101 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     5161 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     5221 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     5281 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     5341 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     5401 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     5461 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     5521 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     5581 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     5641 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     5701 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     5761 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     5821 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     5881 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     5941 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     6001 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     6061 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     6121 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     6181 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     6241 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     6301 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     6361 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     6421 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     6481 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     6541 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     6601 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     6661 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     6721 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     6781 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     6841 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     6901 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     6961 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     7021 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     7081 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     7141 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     7201 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     7261 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     7321 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     7381 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     7441 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     7501 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     7561 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     7621 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     7681 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     7741 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     7801 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     7861 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     7921 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     7981 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     8041 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa
     8101 ggatcttgat tgatcgatag agctcgaatt tccccgatcg ttcaaacatt tggcaataaa
     8161 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     8221 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     8281 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     8341 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg ggatccttcc
     8401 atgggaattc tgagctctct tcatcggtga ttgattcctt taaagactta tgtttcttat
     8461 cttgcttctg aggcaagtat tcagttacca gttaccactt atattctgga ctttctgact
     8521 gcatcctcat ttttccaaca ttttaaattt cactattggc tgaatgcttc ttctttgagg
     8581 aagaaacaat tcagatggca gaaatgtatc aaccaatgca tatatacaaa tgtacctctt
     8641 gttctcaaaa catctatcgg atggttccat ttgctttgtc atccaattag tgactacttt
     8701 atattattca ctcctcttta ttactatttt catgcgaggt tgccatgtac attatatttg
     8761 taaggattga cgctattgag cgtttttctt caattttctt tattttagac atgggtatga
     8821 aatgtgtgtt agagttgggt tgaatgagat atacgttcaa gtgaagtggc ataccgttgt
     8881 cgagtaagga tgacctaccc attcttgaga caaatgttac attttagtat cagagtaaaa
     8941 tgtgtaccta taactcaaat tcgattgaca tgtatccatt caacataaaa ttaaaccagc
     9001 ctgcacctgc atccacattt caagtatttt caaaccgttc ggctcctatc caccgggtgt
     9061 aacaagacgg attccgaatt tggaagattt tgactcaaat tcccaattta tattgaccgt
     9121 gactaaatca actttaactt ctataattct gattaagctc ccaatttata ttcccaacgg
     9181 cactacctcc aaaatttata gactctcatc cccttttaaa ccaacttagt aaacgctctt
     9241 ttttttaatt ttatgaagtt aagtttttac cttgttttta aaaagaatcg ttcataagat
     9301 gccatgccag aacattagct acacgttaca catagcatgc agccgcggag aattgttttt
     9361 cttcgccact tgtcactccc ttcaaacacc taagagcttc tctctcacag cacacacata
     9421 caatcacatg cgtgcatgca ttattacacg tgatcgccat gcaaatctcc tttatagtct
     9481 ataaattaac tcatccgctt cactctttac tcaaaccaaa actcatcaat acaaacaaga
     9541 ttaaaaacat accccatgcg gtcttccaag aatgttatca aggagttcat gaggtttaag
     9601 gttcgcatgg aaggaacggt caatgggcac gagtttgaaa tagaaggcga aggagagggg
     9661 aggccatacg aaggccacaa taccgtaaag cttaaggtaa ccaagggggg acctttgcca
     9721 tttgcttggg atattttgtc accacaattt cagtatggaa gcaaggtata tgtcaagcac
     9781 cctgccgaca taccagacta taaaaagctg tcatttcctg aaggatttaa atgggaaagg
     9841 gtcatgaact ttgaagacgg tggcgtcgtt actgtaaccc aggattccag tttgcaggat
     9901 ggctgtttca tctacaaggt caagttcatt ggcgtgaact ttccttccga tggacctgtt
     9961 atgcaaaaga agacaatggg ctgggaagcc agcactgagc gtttgtatcc tcgtgatggc
    10021 gtgttgaaag gagagattca taaggctctg aagctgaaaa acggtggtca ttacctagtt
    10081 gaattcaaaa gtatttacat ggcaaagaag cctgtgcagc taccagggta ctactatgtt
    10141 gactccaaac tggatataac aagccacaac gaagactata caatcgttga gcagtatgaa
    10201 agaaccgagg gacgccacca tctgttcctt taagatcgtt caaacatttg gcaataaagt
    10261 ttcttaagat tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat
    10321 tacgttaagc atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt
    10381 atgattagag tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca
    10441 aactaggata aattatcgcg cgcggtgtca tctatgttac tagatcgcat gggaattctg
    10501 agatttttca aatcagtgcg caagacgtga cgtaagtatc cgagtcagtt tttatttttc
    10561 tactaatttg gtcgtttatt tcggcgtgta ggacatggca accgggcctg aatttcgcgg
    10621 gtattctgtt tctattccaa ctttttcttg atccgcagcc attaacgact tttgaataga
    10681 tacgctgaca cgccaagcct cgctagtcaa aagtgtacca aacaacgctt tacagcaaga
    10741 acggaatgcg cgtgacgctc gcggtgacgc catttcgcct tttcagaaat ggataaatag
    10801 ccttgcttcc tattatatct tcccaaatta ccaatacatt acactagcat ctgaatttca
    10861 taaccaatct cgatacacca aatcgaatcg atgagcccag aacgacgccc ggccgacatc
    10921 cgccgtgcca ccgaggcgga catgccggcg gtctgcacca tcgtcaacca ctacatcgag
    10981 acaagcacgg tcaacttccg taccgagccg caggaaccgc aggagtggac ggacgacctc
    11041 gtccgtctgc gggagcgcta tccctggctc gtcgccgagg tggacggcga ggtcgccggc
    11101 atcgcctacg cgggcccctg gaaggcacgc aacgcctacg actggacggc cgagtcgacc
    11161 gtgtacgtct ccccccgcca ccagcggacg ggactgggct ccacgctcta cacccacctg
    11221 ctgaagtccc tggaggcaca gggcttcaag agcgtggtcg ctgtcatcgg gctgcccaac
    11281 gacccgagcg tgcgcatgca cgaggcgctc ggatatgccc cccgcggcat gctgcgggcg
    11341 gccggcttca agcacgggaa ctggcatgac gtgggtttct ggcagctgga cttcagcctg
    11401 ccggtaccgc cccgtccggt cctgcccgtc accgagattt gactgcagaa tcttggactc
    11461 ccatgttggc aaaggcaacc aaacaaacaa tgaatgatcc gctcctgcat atggggcggt
    11521 ttgagtattt caactgccat ttgggctgaa ttgaagacat gctcctgtca gaaattccgt
    11581 gatcttactc aatattcagt aatctcggcc aatatcctaa atgtgcgtgg ctttatctgt
    11641 ctttgtattg tttcatcaat tcatgtaacg tttgcttttc ttatgaattt tcaaataaat
    11701 tatcagatcc gagctcgaat taattcggcg ttaattcagt acattaaaaa cgtccgcaat
    11761 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
    11821 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
    11881 tcagtccggg acggcgtcag cgggagagcc gttgtaaggc ggcagacttt gctcatgtta
    11941 ccgatgctat tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc
    12001 ggagggtagc atgttgattg taacgatgac agagcgttgc tgcctgtgat caccgcggtt
    12061 tcaaaatcgg ctccgtcgat actatgttat acgccaactt tgaaaacaac tttgaaaaag
    12121 ctgttttctg gtatttaagg ttttagaatg caaggaacag tgaattggag ttcgtcttgt
    12181 tataattagc ttcttggggt atctttaaat actgtagaaa agaggaagga aataataaat
    12241 ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa
    12301 agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct
    12361 atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa
    12421 ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg
    12481 gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga
    12541 gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct
    12601 ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc
    12661 cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga
    12721 agacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga
    12781 agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg
    12841 caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat
    12901 tgccttctgc gtccggtcga tcagggagga tatcggggaa gaacagtatg tcgagctatt
    12961 ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga
    13021 tgaattgttt tagtacctag aatgcatgac caaaatccct taacgtgagt tttcgttcca
    13081 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    13141 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    13201 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    13261 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    13321 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
    13381 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
    13441 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
    13501 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
    13561 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
    13621 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
    13681 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
    13741 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
    13801 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
    13861 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
    13921 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
    13981 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
    14041 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
    14101 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
    14161 aaacgcgcga ggcagggtgc cttgatgtgg gcgccggcgg tcgagtggcg acggcgcggc
    14221 ttgtccgcgc cctggtagat tgcctggccg taggccagcc atttttgagc ggccagcggc
    14281 cgcgataggc cgacgcgaag cggcggggcg tagggagcgc agcgaccgaa gggtaggcgc
    14341 tttttgcagc tcttcggctg tgcgctggcc agacagttat gcacaggcca ggcgggtttt
    14401 aagagtttta ataagtttta aagagtttta ggcggaaaaa tcgccttttt tctcttttat
    14461 atcagtcact tacatgtgtg accggttccc aatgtacggc tttgggttcc caatgtacgg
    14521 gttccggttc ccaatgtacg gctttgggtt cccaatgtac gtgctatcca caggaaagag
    14581 accttttcga cctttttccc ctgctagggc aatttgccct agcatctgct ccgtacatta
    14641 ggaaccggcg gatgcttcgc cctcgatcag gttgcggtag cgcatgacta ggatcgggcc
    14701 agcctgcccc gcctcctcct tcaaatcgta ctccggcagg tcatttgacc cgatcagctt
    14761 gcgcacggtg aaacagaact tcttgaactc tccggcgctg ccactgcgtt cgtagatcgt
    14821 cttgaacaac catctggctt ctgccttgcc tgcggcgcgg cgtgccaggc ggtagagaaa
    14881 acggccgatg ccgggatcga tcaaaaagta atcggggtga accgtcagca cgtccgggtt
    14941 cttgccttct gtgatctcgc ggtacatcca atcagctagc tcgatctcga tgtactccgg
    15001 ccgcccggtt tcgctcttta cgatcttgta gcggctaatc aaggcttcac cctcggatac
    15061 cgtcaccagg cggccgttct tggccttctt cgtacgctgc atggcaacgt gcgtggtgtt
    15121 taaccgaatg caggtttcta ccaggtcgtc tttctgcttt ccgccatcgg ctcgccggca
    15181 gaacttgagt acgtccgcaa cgtgtggacg gaacacgcgg ccgggcttgt ctcccttccc
    15241 ttcccggtat cggttcatgg attcggttag atgggaaacc gccatcagta ccaggtcgta
    15301 atcccacaca ctggccatgc cggccggccc tgcggaaacc tctacgtgcc cgtctggaag
    15361 ctcgtagcgg atcacctcgc cagctcgtcg gtcacgcttc gacagacgga aaacggccac
    15421 gtccatgatg ctgcgactat cgcgggtgcc cacgtcatag agcatcggaa cgaaaaaatc
    15481 tggttgctcg tcgcccttgg gcggcttcct aatcgacggc gcaccggctg ccggcggttg
    15541 ccgggattct ttgcggattc gatcagcggc cgcttgccac gattcaccgg ggcgtgcttc
    15601 tgcctcgatg cgttgccgct gggcggcctg cgcggccttc aacttctcca ccaggtcatc
    15661 acccagcgcc gcgccgattt gtaccgggcc ggatggtttg cgaccgctca cgccgattcc
    15721 tcgggcttgg gggttccagt gccattgcag ggccggcagg caacccagcc gcttacgcct
    15781 ggccaaccgc ccgttcctcc acacatgggg cattccacgg cgtcggtgcc tggttgttct
    15841 tgattttcca tgccgcctcc tttagccgct aaaattcatc tactcattta ttcatttgct
    15901 catttactct ggtagctgcg cgatgtattc agatagcagc tcggtaatgg tcttgccttg
    15961 gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc ggcaactgaa agttgacccg
    16021 cttcatggct ggcgtgtctg ccaggctggc caacgttgca gccttgctgc tgcgtgcgct
    16081 cggacggccg gcacttagcg tgtttgtgct tttgctcatt ttctctttac ctcattaact
    16141 caaatgagtt ttgatttaat ttcagcggcc agcgcctgga cctcgcgggc agcgtcgccc
    16201 tcgggttctg attcaagaac ggttgtgccg gcggcggcag tgcctgggta gctcacgcgc
    16261 tgcgtgatac gggactcaag aatgggcagc tcgtacccgg ccagcgcc
//
