LOCUS       sequence-404047-       16211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Cas9 in plants, cauliflower mosaic virus 35S promoter 
            (P35S), Neo selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16211)
  AUTHORS   Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, 
            Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X,
            Dong Z, Liu YG
  TITLE     A robust CRISPR/Cas9 system for convenient, high-efficiency 
            multiplex genome editing in monocot and dicot plants.
  JOURNAL   Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 
            10.1016/j.molp.2015.04.007.
  PUBMED    25917172
REFERENCE   2  (bases 1 to 16211)
  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: "Mol Plant. 
            2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 
            10.1016/j.molp.2015.04.007."
COMMENT     Sequence Label: pYLCRISPR/Cas9P35S-N
FEATURES             Location/Qualifiers
     source          1..16211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        260..604
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             753..773
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             798..4898
                     /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             798..4898
                     /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"
     RBS             1984..1992
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4899..4946
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5287..5303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5311..5327
                     /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(5335..5341,5342..5359,5360..5365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5380..5401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5592..6268
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             6331..7128
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6388..6407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6998..7017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7185..7359
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    7437..7461
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             9085..9879
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9966..10554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10455..10474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10708..10725
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    10740..10880
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10806..10825)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10966..10988
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      11224..11418
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11484..12557)
                     /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(12986..13615)
                     /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    14916..14940
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     15128..15150
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15142..15159
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15143..15159
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(15189..15222)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(15432..15734)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(15534..15553)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        complement(15808..15826)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(15809..15826)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    15868..15884
                     /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(15892..15898,15899..15916,15917..15922))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15937..15958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttattgtgaa gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag
       61 gaaaggccat cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
      121 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
      181 gtgataacat ggtggagcac gacacacttg tctactccaa aaatatcaaa gatacagtct
      241 cagaagacca aagggcaatt gagacttttc aacaaagggt aatatccgga aacctcctcg
      301 gattccattg cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct
      361 cctacaaatg ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca
      421 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
      481 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
      541 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
      601 ggacgtcgag agttctcaac acaacatata caaaacaaac gaatctcaag caatcaagca
      661 ttctacttct attgcagcaa tttaaatcat ttcttttaaa gcaaaagcaa ttttctgaaa
      721 attttcacca tttacgaacg atagccatgg ctcctaagaa gaagcggaag gttggtattc
      781 acggggtgcc tgcggctgac aagaagtact ccatcggcct cgacatcggc accaacagcg
      841 tcggctgggc ggtgatcacc gacgagtaca aggtcccgtc caagaagttc aaggtcctgg
      901 gcaacaccga ccgccactcc atcaagaaga acctcatcgg cgccctcctc ttcgactccg
      961 gcgagacggc ggaggcgacc cgcctcaagc gcaccgcccg ccgccgctac acccgccgca
     1021 agaaccgcat ctgctacctc caggagatct tctccaacga gatggcgaag gtcgacgact
     1081 ccttcttcca ccgcctcgag gagtccttcc tcgtggagga ggacaagaag cacgagcgcc
     1141 accccatctt cggcaacatc gtcgacgagg tcgcctacca cgagaagtac cccactatct
     1201 accaccttcg taagaagctt gttgactcta ctgataaggc tgatcttcgt ctcatctacc
     1261 ttgctctcgc tcacatgatc aagttccgtg gtcacttcct tatcgagggt gaccttaacc
     1321 ctgataactc cgacgtggac aagctcttca tccagctcgt ccagacctac aaccagctct
     1381 tcgaggagaa ccctatcaac gcttccggtg tcgacgctaa ggcgatcctt tccgctaggc
     1441 tctccaagtc caggcgtctc gagaacctca tcgcccagct ccctggtgag aagaagaacg
     1501 gtcttttcgg taacctcatc gctctctccc tcggtctgac ccctaacttc aagtccaact
     1561 tcgacctcgc tgaggacgct aagcttcagc tctccaagga tacctacgac gatgatctcg
     1621 acaacctcct cgctcagatt ggagatcagt acgctgatct cttccttgct gctaagaacc
     1681 tctccgatgc tatcctcctt tcggatatcc ttagggttaa cactgagatc actaaggctc
     1741 ctctttctgc ttccatgatc aagcgctacg acgagcacca ccaggacctc accctcctca
     1801 aggctcttgt tcgtcagcag ctccccgaga agtacaagga gatcttcttc gaccagtcca
     1861 agaacggcta cgccggttac attgacggtg gagctagcca ggaggagttc tacaagttca
     1921 tcaagccaat ccttgagaag atggatggta ctgaggagct tctcgttaag cttaaccgtg
     1981 aggacctcct taggaagcag aggactttcg ataacggctc tatccctcac cagatccacc
     2041 ttggtgagct tcacgccatc cttcgtaggc aggaggactt ctaccctttc ctcaaggaca
     2101 accgtgagaa gatcgagaag atccttactt tccgtattcc ttactacgtt ggtcctcttg
     2161 ctcgtggtaa ctcccgtttc gcttggatga ctaggaagtc cgaggagact atcacccctt
     2221 ggaacttcga ggaggttgtt gacaagggtg cttccgccca gtccttcatc gagcgcatga
     2281 ccaacttcga caagaacctc cccaacgaga aggtcctccc caagcactcc ctcctctacg
     2341 agtacttcac ggtctacaac gagctcacca aggtcaagta cgtcaccgag ggtatgcgca
     2401 agcctgcctt cctctccggc gagcagaaga aggctatcgt tgacctcctc ttcaagacca
     2461 accgcaaggt caccgtcaag cagctcaagg aggactactt caagaagatc gagtgcttcg
     2521 actccgtcga gatcagcggc gttgaggacc gtttcaacgc ttctctcggt acctaccacg
     2581 atctcctcaa gatcatcaag gacaaggact tcctcgacaa cgaggagaac gaggacatcc
     2641 tcgaggacat cgtcctcact cttactctct tcgaggatag ggagatgatc gaggagaggc
     2701 tcaagactta cgctcatctc ttcgatgaca aggttatgaa gcagctcaag cgtcgccgtt
     2761 acaccggttg gggtaggctc tcccgcaagc tcatcaacgg tatcagggat aagcagagcg
     2821 gcaagactat cctcgacttc ctcaagtctg atggtttcgc taacaggaac ttcatgcagc
     2881 tcatccacga tgactctctt accttcaagg aggatattca gaaggctcag gtgtccggtc
     2941 agggcgactc tctccacgag cacattgcta accttgctgg ttcccctgct atcaagaagg
     3001 gcatccttca gactgttaag gttgtcgatg agcttgtcaa ggttatgggt cgtcacaagc
     3061 ctgagaacat cgtcatcgag atggctcgtg agaaccagac tacccagaag ggtcagaaga
     3121 actcgaggga gcgcatgaag aggattgagg agggtatcaa ggagcttggt tctcagatcc
     3181 ttaaggagca ccctgtcgag aacacccagc tccagaacga gaagctctac ctctactacc
     3241 tccagaacgg tagggatatg tacgttgacc aggagctcga catcaacagg ctttctgact
     3301 acgacgtcga ccacattgtt cctcagtctt tccttaagga tgactccatc gacaacaagg
     3361 tcctcacgag gtccgacaag aacaggggta agtcggacaa cgtcccttcc gaggaggttg
     3421 tcaagaagat gaagaactac tggaggcagc ttctcaacgc taagctcatt acccagagga
     3481 agttcgacaa cctcacgaag gctgagaggg gtggcctttc cgagcttgac aaggctggtt
     3541 tcatcaagag gcagcttgtt gagacgaggc agattaccaa gcacgttgct cagatcctcg
     3601 attctaggat gaacaccaag tacgacgaga acgacaagct catccgcgag gtcaaggtga
     3661 tcaccctcaa gtccaagctc gtctccgact tccgcaagga cttccagttc tacaaggtcc
     3721 gcgagatcaa caactaccac cacgctcacg atgcttacct taacgctgtc gttggtaccg
     3781 ctcttatcaa gaagtaccct aagcttgagt ccgagttcgt ctacggtgac tacaaggtct
     3841 acgacgttcg taagatgatc gccaagtccg agcaggagat cggcaaggcc accgccaagt
     3901 acttcttcta ctccaacatc atgaacttct tcaagaccga gatcaccctc gccaacggcg
     3961 agatccgcaa gcgccctctt atcgagacga acggtgagac tggtgagatc gtttgggaca
     4021 agggtcgcga cttcgctact gttcgcaagg tcctttctat gcctcaggtt aacatcgtca
     4081 agaagaccga ggtccagacc ggtggcttct ccaaggagtc tatccttcca aagagaaact
     4141 cggacaagct catcgctagg aagaaggatt gggaccctaa gaagtacggt ggtttcgact
     4201 cccctactgt cgcctactcc gtcctcgtgg tcgccaaggt ggagaagggt aagtcgaaga
     4261 agctcaagtc cgtcaaggag ctcctcggca tcaccatcat ggagcgctcc tccttcgaga
     4321 agaacccgat cgacttcctc gaggccaagg gctacaagga ggtcaagaag gacctcatca
     4381 tcaagctccc caagtactct cttttcgagc tcgagaacgg tcgtaagagg atgctggctt
     4441 ccgctggtga gctccagaag ggtaacgagc ttgctcttcc ttccaagtac gtgaacttcc
     4501 tctacctcgc ctcccactac gagaagctca agggttcccc tgaggataac gagcagaagc
     4561 agctcttcgt ggagcagcac aagcactacc tcgacgagat catcgagcag atctccgagt
     4621 tctccaagcg cgtcatcctc gctgacgcta acctcgacaa ggtcctctcc gcctacaaca
     4681 agcaccgcga caagcccatc cgcgagcagg ccgagaacat catccacctc ttcacgctca
     4741 cgaacctcgg cgcccctgct gctttcaagt acttcgacac caccatcgac aggaagcgtt
     4801 acacgtccac caaggaggtt ctcgacgcta ctctcatcca ccagtccatc accggtcttt
     4861 acgagactcg tatcgacctt tcccagcttg gtggtgataa gcgtcctgct gccaccaaaa
     4921 aggccggaca ggctaagaaa aagaagtagg atccacctga tctagagtcc gcaaaaatca
     4981 ccagtctctc tctacaaatc tatctctctc tatttttctc cagaataatg tgtgagtagt
     5041 tcccagataa gggaattagg gttcttatag ggtttcgctc atgtgttgag catataagaa
     5101 acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa
     5161 aaccaaaatc cagtgacgcg gccgcataag gccggccttg ccctagttcc tatagcctac
     5221 attataggat ggagggatat cctctcttaa ggtagcgagc ggtaccgagc tcgaattcgt
     5281 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggctagagca gcttgccaac
     5521 atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac
     5581 caaagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat
     5641 tgcccagcta tctgtcactt catcaaaagg acagtagaaa aggaaggtgg cacctacaaa
     5701 tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctgccga cagtggtccc
     5761 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
     5821 tcaaagcaag tggattgatg tgaacatggt ggagcacgac actctcgtct actccaagaa
     5881 tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac aaagggtaat
     5941 atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca aaaggacagt
     6001 agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca
     6061 agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgaaga gcatcgtgga
     6121 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
     6181 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
     6241 ttcatttcat ttggagagga cacgctgaaa tcaccagtct ctctctacaa atctatctct
     6301 ctcgagcttt cgcagatctg tcgatcgacc atggggattg aacaagatgg attgcacgca
     6361 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6421 ggctgctctg atgccgccgt gtttcggctg tcagcgcagg ggcgcccggt tctttttgtc
     6481 aagaccgacc tgtccggtgc cctgaatgaa ctccaggacg aggcagcgcg gctatcgtgg
     6541 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6601 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6661 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6721 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6781 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6841 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacacatggc
     6901 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6961 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     7021 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     7081 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     7141 ggttcggatc gatcctctag ctagagtcga tcgacaagct cgagtttctc cataataatg
     7201 tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc atgtgttgag
     7261 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
     7321 taattcctaa aaccaaaatc cagtactaaa atccagatcc cccgaattaa ttcggcgtta
     7381 attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt
     7441 acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca
     7501 aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg gagagccgtt
     7561 gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc aactaagctg
     7621 ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa cgatgacaga
     7681 gcgttgctgc ctgtgatcac cgcggtttca aaatcggctc cgtcgatact atgttatacg
     7741 ccaactttga aaacaacttt gaaaaagctg ttttctggta tttaaggttt tagaatgcaa
     7801 ggaacagtga attggagttc gtcttgttat aattagggaa ggtgcgaata agcggggaaa
     7861 ttcttctcgg ctgactcagt catttcattt cttcatgttt gagccgattt tttctcccgt
     7921 aaatgccttg aatcagccta tttagaccgt ttcttcgcca tttaaggcgt tatccccagt
     7981 ttttagtgag atctctccca ctgacgtatc atttggtccg cccgaaacag gttggccagc
     8041 gtgaataaca tcgccagttg gttatcgttt ttcagcaacc ccttgtatct ggctttcacg
     8101 aagccgaact gtcgcttgat gatgcgaaat gggtgctcca ccctggcccg gatgctggct
     8161 ttcatgtatt cgatgttgat ggccgttttg ttcttgcgtg gatgctgttt caaggttctt
     8221 accttgccgg ggcgctcggc gatcagccag tccacatcca cctcggccag ctcctcgcgc
     8281 tgtggcgccc cttggtagcc ggcatcggct gagacaaatt gctcctctcc atgcagcaga
     8341 ttacccagct gattgaggtc atgctcgttg gccgcggtgg tgaccaggct gtgggtcagg
     8401 ccactcttgg catcgacacc aatgtgggcc ttcatgccaa agtgccactg attgcctttc
     8461 ttggtctgat gcatctccgg atcgcgttgc tgctctttgt tcttggtcga gctgggtgcc
     8521 tcaatgatgg tggcatcgac caaggtgcct tgagtcatca tgacgcctgc ttcggccagc
     8581 cagcgattga tggtcttgaa caattggcgg gccagttgat gctgctccag caggtggcgg
     8641 aaattcatga tggtggtgcg gtccggcaag gcgctatcca gggataaccg ggcaaacaga
     8701 cgcatggagg cgatttcgta cagagcatct tccatcgcgc catcgctcag gttgtaccaa
     8761 tgctgcatgc agtgaatgcg tagcatggtt tccagcggat aaggtcgccg gccattacca
     8821 gccttggggt aaaacggctc gatgacttcc accatgtttt gccatggcag aatctgctcc
     8881 atgcgggaca agaaaatctc ttttctggtc tgacggcgct tactgctgaa ttcactgtcg
     8941 gcgaaggtaa gttgatgact catgatgaac cctgttctat ggctccagat gacaaacatg
     9001 atctcatatc agggacttgt tcgcaccttc cttagcttct tggggtatct ttaaatactg
     9061 tagaaaagag gaaggaaata ataaatggct aaaatgagaa tatcaccgga attgaaaaaa
     9121 ctgatcgaaa aataccgctg cgtaaaagat acggaaggaa tgtctcctgc taaggtatat
     9181 aagctggtgg gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg
     9241 accacctatg atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct
     9301 gttccaaagg tcctgcactt tgaacggcat gatggctgga gcaatctgct catgagtgag
     9361 gccgatggcg tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc
     9421 gagctgtatg cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat
     9481 acgaatagct tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc
     9541 gatgtggatt gcgaaaactg ggaagaagac actccattta aagatccgcg cgagctgtat
     9601 gattttttaa agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga
     9661 gacagcaaca tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc
     9721 ggcagggcgg acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc
     9781 ggggaagaac agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag
     9841 aaaataaaat attatatttt actggatgaa ttgttttagt acctagaatg catgaccaaa
     9901 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     9961 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    10021 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    10081 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
    10141 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    10201 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    10261 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    10321 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    10381 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    10441 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    10501 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    10561 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    10621 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
    10681 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
    10741 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
    10801 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
    10861 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
    10921 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
    10981 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc
    11041 cggcggtcga gtggcgacgg cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg
    11101 ccagccattt ttgagcggcc agcggccgcg ataggccgac gcgaagcggc ggggcgtagg
    11161 gagcgcagcg accgaagggt aggcgctttt tgcagctctt cggctgtgcg ctggccagac
    11221 agttatgcac aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg
    11281 gaaaaatcgc cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg
    11341 tacggctttg ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca
    11401 atgtacgtgc tatccacagg aaagagaact tttcgacctt tttcccctgc tagggcaatt
    11461 tgccctagca tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg
    11521 cggtagcgca tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc
    11581 ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg
    11641 gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg
    11701 gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg
    11761 gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca
    11821 gctagctcga tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg
    11881 ctaatcaagg cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta
    11941 cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc
    12001 tgctttccgc catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac
    12061 acgcggccgg gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg
    12121 gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg
    12181 gaaacctcta cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca
    12241 cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg
    12301 tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc
    12361 gacggcgcac cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct
    12421 tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg
    12481 gccttcaact tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat
    12541 ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc
    12601 ggcaggcaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt
    12661 ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa
    12721 ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat gtattcagat
    12781 agcagctcgg taatggtctt gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc
    12841 tccgccggca actgaaagtt gacccgcttc atggctggcg tgtctgccag gctggccaac
    12901 gttgcagcct tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg
    12961 ctcattttct ctttacctca ttaactcaaa tgagttttga tttaatttca gcggccagcg
    13021 cctggacctc gcgggcagcg tcgccctcgg gttctgattc aagaacggtt gtgccggcgg
    13081 cggcagtgcc tgggtagctc acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt
    13141 acccggccag cgcctcggca acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca
    13201 cgacaaaggc cgcttgtagc cttccatccg tgacctcaat gcgctgctta accagctcca
    13261 ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg caccggaatc agcacgaagt
    13321 cggctgcctt gatcgcggac acagccaagt ccgccgcctg gggcgctccg tcgatcacta
    13381 cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga
    13441 caacggttag cggttgatct tcccgcacgg ccgcccaatc gcgggcactg ccctggggat
    13501 cggaatcgac taacagaaca tcggccccgg cgagttgcag ggcgcgggct agatgggttg
    13561 cgatggtcgt cttgcctgac ccgcctttct ggttaagtac agcgataacc ttcatgcgtt
    13621 ccccttgcgt atttgtttat ttactcatcg catcatatac gcagcgaccg catgacgcaa
    13681 gctgttttac tcaaatacac atcacctttt tagacggcgg cgctcggttt cttcagcggc
    13741 caagctggcc ggccaggccg ccagcttggc atcagacaaa ccggccagga tttcatgcag
    13801 ccgcacggtt gagacgtgcg cgggcggctc gaacacgtac ccggccgcga tcatctccgc
    13861 ctcgatctct tcggtaatga aaaacggttc gtcctggccg tcctggtgcg gtttcatgct
    13921 tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc
    13981 tcacggaagg caccgcgccg cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt
    14041 gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg
    14101 ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg
    14161 gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg
    14221 attagggaac gctcgaactc ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc
    14281 ggcgtggtgg tgtcggccca cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg
    14341 atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc
    14401 actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg
    14461 cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc agccggccgc gtgcagttcg
    14521 gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca
    14581 gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct agtcgcaagt attctacttt
    14641 atgcgactaa aacacgcgac aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc
    14701 gtaacttagg acttgtgcga catgtcgttt tcagaagacg gctgcactga acgtcagaag
    14761 ccgactgcac tatagcagcg gaggggttgg atcaaagtac tttgatcccg aggggaaccc
    14821 tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat
    14881 atccgattat tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca
    14941 aacactgata gtttaaactg aaggcgggaa acgacaatct gatccaagct caagctgctc
    15001 tagcattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
    15061 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
    15121 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagcttat gggtgtcgac
    15181 ttgcagtcat aacttcgtat agcatacatt atacgaagtt atcagatcta atactgccat
    15241 ttcattacct ctttctccgc acccgacata gatgcaataa cttcgtatag gctaatttat
    15301 acgatgtagg agatcgatgc atgcggccgc tagctcgaga ggcgcgccaa tgataccgcg
    15361 agacccacgc tcacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
    15421 aggcctgaca tttatattcc ccagaacatc aggttaatgg cgtttttgat gtcattttcg
    15481 cggtggctga gatcagccac ttcttccccg ataacggaga caggcacact ggccatatcg
    15541 gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag ttcacgggag
    15601 actttatctg acagcagacg tgcactggcc agggggatca ccatccgtcg ccccggcgtg
    15661 tcaataatat cactctgtac atccacaaac agacgataac ggctctctct tttataggtg
    15721 taaaccttaa actgatgttc gagcggtcgc cagtgtgatg ggtatctaca gactagaccc
    15781 attgagcttg atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
    15841 atggtcatag ctgtttcttg tgtgaaattg ttatccgctc acaattccac acaacatacg
    15901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    15961 cgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    16021 aatcggccac ttcagaggtc tctcgagcac tacggcgcgc cttaccggtc aacatgtgga
    16081 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
    16141 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
    16201 tatctgtcac t
//
