LOCUS       sequence-112919-       15889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Cas9 in plants, maize ubiquitin promoter (Pubi), basta
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15889)
  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 15889)
  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/Cas9Pubi-B
FEATURES             Location/Qualifiers
     source          1..15889
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    103..277
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(284..835)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(880..1557)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    1748..1769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1784..1789,1790..1807,1808..1814)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1822..1838
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1846..1862
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4003..4023
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4048..8148
                     /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             4048..8148
                     /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             5234..5242
                     /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             8149..8196
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8209..8461
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8579..8600
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8615..8620,8621..8638,8639..8645)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8653..8669
                     /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        8711..8729
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     8711..8728
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             8803..9105
                     /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     8984..9003
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    9315..9348
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9612..9636
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10936..11565
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             11999..13066
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      13132..13326
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13562..13584)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13670..13810
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     13725..13744
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(13825..13842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13996..14584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14076..14095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14671..15465)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa ttaacgccga attaattcgg gggatctgga ttttagtact
      121 ggattttggt tttaggaatt agaaatttta ttgatagaag tattttacaa atacaaatac
      181 atactaaggg tttcttatat gctcaacaca tgagcgaaac cctataggaa ccctaattcc
      241 cttatctggg aactactcac acattattat ggagaaactc gagtcaaatc tcggtgacgg
      301 gcaggaccgg acggggcggt accggcaggc tgaagtccag ctgccagaaa cccacgtcat
      361 gccagttccc gtgcttgaag ccggccgccc gcagcatgcc gcggggggca tatccgagcg
      421 cctcgtgcat gcgcacgctc gggtcgttgg gcagcccgat gacagcgacc acgctcttga
      481 agccctgtgc ctccagggac ttcagcaggt gggtgtagag cgtggagccc agtcccgtcc
      541 gctggtggcg gggggagacg tacacggtcg actcggccgt ccagtcgtag gcgttgcgtg
      601 ccttccaggg gcccgcgtag gcgatgccgg cgacctcgcc gtccacctcg gcgacgagcc
      661 agggatagcg ctcccgcaga cggacgaggt cgtccgtcca ctcctgcggt tcctgcggct
      721 cggtacggaa gttgaccgtg cttgtctcga tgtagtggtt gacgatggtg cagaccgccg
      781 gcatgtccgc ctcggtggca cggcggatgt cggccgggcg tcgttctggg ctcatggtag
      841 actcgagaga gatagatttg tagagagaga ctggtgattt cagcgtgtcc tctccaaatg
      901 aaatgaactt ccttatatag aggaaggtct tgcgaaggat agtgggattg tgcgtcatcc
      961 cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct
     1021 tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag
     1081 gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc
     1141 ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt
     1201 cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct
     1261 ttgatattct tggagtagac gagagtgtcg tgctccacca tgttatcaca tcaatccact
     1321 tgctttgaag acgtggttgg aacgtcttct ttttccacga tgctcctcgt gggtgggggt
     1381 ccatctttgg gaccactgtc ggcagaggca tcttgaacga tagcctttcc tttatcgcaa
     1441 tgatggcatt tgtaggtgcc accttccttt tctactgtcc ttttgatgaa gtgacagata
     1501 gctgggcaat ggaatccgag gaggtttccc gatattaccc tttgttgaaa agtctcaata
     1561 gccctttggt cttctgagac tgtatctttg atattcttgg agtagacgag agtgtcgtgc
     1621 tccaccatgt tggcaagctg ctctagccaa tacgcaaacc gcctctcccc gcgcgttggc
     1681 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1741 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1801 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1861 accatgatta cgaattcgag ctcggtacca actataacgg tcctaaggta gcgaaggatc
     1921 cgctcgctac cttaagagag gatatccctc catcctataa tgtaggctat aggaactagg
     1981 gcaaggccgg ccatgcggcc gcaagctggg tgcagcgtga cccggtcgtg cccctctcta
     2041 gagataatga gcattgcatg tctaagttat aaaaaattac cacatatttt ttttgtcaca
     2101 cttgtttgaa gtgcagttta tctatcttta tacatatatt taaactttac tctacgaata
     2161 atataatcta tagtactaca ataatatcag tgttttagag aatcatataa atgaacagtt
     2221 agacatggtc taaaggacaa ttgagtattt tgacaacagg actctacagt tttatctttt
     2281 tagtgtgcat gtgttctcct ttttttttgc aaatagcttc acctatataa tacttcatcc
     2341 attttattag tacatccatt tagggtttag ggttaatggt ttttatagac taattttttt
     2401 agtacatcta ttttattcta ttttagcctc taaattaaga aaactaaaac tctattttag
     2461 tttttttatt taataattta gatataaaat agaataaaat aaagtgacta aaaattaaac
     2521 aaataccctt taagaaatta aaaaaactaa ggaaacattt ttcttgtttc gagtagataa
     2581 tgccagcctg ttaaacgccg tcgacgagtc taacggacac caaccagcga accagcagcg
     2641 tcgcgtcggg ccaagcgaag cagacggcac ggcatctctg tcgctgcctc tggacccctc
     2701 tcgagagttc cgctccaccg ttggacttgc tccgctgtcg gcatccagaa attgcgtggc
     2761 ggagcggcag acgtgagccg gcacggcagg cggcctcctc ctcctctcac ggcacggcag
     2821 ctacggggga ttcctttccc accgctcctt cgctttccct tcctcgcccg ccgtaataaa
     2881 tagacacccc ctccacaccc tctttcccca acctcgtgtt gttcggagcg cacacacaca
     2941 caaccagatc tcccccaaat ccacccgtcg gcacctccgc ttcaaggtac gccgctcgtc
     3001 ctcccccccc ccccctctct accttctcta gatcggcgtt ccggtccatg gttagggccc
     3061 ggtagttcta cttctgttca tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc
     3121 tagcgttcgt acacggatgc gacctgtacg tcagacacgt tctgattgct aacttgccag
     3181 tgtttctctt tggggaatcc tgggatggct ctagccgttc cgcagacggg atcgatttca
     3241 tgattttttt tgtttcgttg catagggttt ggtttgccct tttcctttat ttcaatatat
     3301 gccgtgcact tgtttgtcgg gtcatctttt catgcttttt tttgtcttgg ttgtgatgat
     3361 gtggtctggt tgggcggtcg ttctagatcg gagtagaatt ctgtttcaaa ctacctggtg
     3421 gatttattaa ttttggatct gtatgtgtgt gccatacata ttcatagtta cgaattgaag
     3481 atgatggatg gaaatatcga tctaggatag gtatacatgt tgatgcgggt tttactgatg
     3541 catatacaga gatgcttttt gttcgcttgg ttgtgatgat gtggtgtggt tgggcggtcg
     3601 ttcattcgtt ctagatcgga gtagaatact gtttcaaact acctggtgta tttattaatt
     3661 ttggaactgt atgtgtgtgt catacatctt catagttacg agtttaagat ggatggaaat
     3721 atcgatctag gataggtata catgttgatg tgggttttac tgatgcatat acatgatggc
     3781 atatgcagca tctattcata tgctctaacc ttgagtacct atctattata ataaacaagt
     3841 atgttttata attattttga tcttgatata cttggatgat ggcatatgca gcagctatat
     3901 gtggattttt ttagccctgc cttcatacgc tatttatttg cttggtactg tttcttttgt
     3961 cgatgctcac cctgttgttt ggtgttactt ctgcagatgg ctcctaagaa gaagcggaag
     4021 gttggtattc acggggtgcc tgcggctgac aagaagtact ccatcggcct cgacatcggc
     4081 accaacagcg tcggctgggc ggtgatcacc gacgagtaca aggtcccgtc caagaagttc
     4141 aaggtcctgg gcaacaccga ccgccactcc atcaagaaga acctcatcgg cgccctcctc
     4201 ttcgactccg gcgagacggc ggaggcgacc cgcctcaagc gcaccgcccg ccgccgctac
     4261 acccgccgca agaaccgcat ctgctacctc caggagatct tctccaacga gatggcgaag
     4321 gtcgacgact ccttcttcca ccgcctcgag gagtccttcc tcgtggagga ggacaagaag
     4381 cacgagcgcc accccatctt cggcaacatc gtcgacgagg tcgcctacca cgagaagtac
     4441 cccactatct accaccttcg taagaagctt gttgactcta ctgataaggc tgatcttcgt
     4501 ctcatctacc ttgctctcgc tcacatgatc aagttccgtg gtcacttcct tatcgagggt
     4561 gaccttaacc ctgataactc cgacgtggac aagctcttca tccagctcgt ccagacctac
     4621 aaccagctct tcgaggagaa ccctatcaac gcttccggtg tcgacgctaa ggcgatcctt
     4681 tccgctaggc tctccaagtc caggcgtctc gagaacctca tcgcccagct ccctggtgag
     4741 aagaagaacg gtcttttcgg taacctcatc gctctctccc tcggtctgac ccctaacttc
     4801 aagtccaact tcgacctcgc tgaggacgct aagcttcagc tctccaagga tacctacgac
     4861 gatgatctcg acaacctcct cgctcagatt ggagatcagt acgctgatct cttccttgct
     4921 gctaagaacc tctccgatgc tatcctcctt tcggatatcc ttagggttaa cactgagatc
     4981 actaaggctc ctctttctgc ttccatgatc aagcgctacg acgagcacca ccaggacctc
     5041 accctcctca aggctcttgt tcgtcagcag ctccccgaga agtacaagga gatcttcttc
     5101 gaccagtcca agaacggcta cgccggttac attgacggtg gagctagcca ggaggagttc
     5161 tacaagttca tcaagccaat ccttgagaag atggatggta ctgaggagct tctcgttaag
     5221 cttaaccgtg aggacctcct taggaagcag aggactttcg ataacggctc tatccctcac
     5281 cagatccacc ttggtgagct tcacgccatc cttcgtaggc aggaggactt ctaccctttc
     5341 ctcaaggaca accgtgagaa gatcgagaag atccttactt tccgtattcc ttactacgtt
     5401 ggtcctcttg ctcgtggtaa ctcccgtttc gcttggatga ctaggaagtc cgaggagact
     5461 atcacccctt ggaacttcga ggaggttgtt gacaagggtg cttccgccca gtccttcatc
     5521 gagcgcatga ccaacttcga caagaacctc cccaacgaga aggtcctccc caagcactcc
     5581 ctcctctacg agtacttcac ggtctacaac gagctcacca aggtcaagta cgtcaccgag
     5641 ggtatgcgca agcctgcctt cctctccggc gagcagaaga aggctatcgt tgacctcctc
     5701 ttcaagacca accgcaaggt caccgtcaag cagctcaagg aggactactt caagaagatc
     5761 gagtgcttcg actccgtcga gatcagcggc gttgaggacc gtttcaacgc ttctctcggt
     5821 acctaccacg atctcctcaa gatcatcaag gacaaggact tcctcgacaa cgaggagaac
     5881 gaggacatcc tcgaggacat cgtcctcact cttactctct tcgaggatag ggagatgatc
     5941 gaggagaggc tcaagactta cgctcatctc ttcgatgaca aggttatgaa gcagctcaag
     6001 cgtcgccgtt acaccggttg gggtaggctc tcccgcaagc tcatcaacgg tatcagggat
     6061 aagcagagcg gcaagactat cctcgacttc ctcaagtctg atggtttcgc taacaggaac
     6121 ttcatgcagc tcatccacga tgactctctt accttcaagg aggatattca gaaggctcag
     6181 gtgtccggtc agggcgactc tctccacgag cacattgcta accttgctgg ttcccctgct
     6241 atcaagaagg gcatccttca gactgttaag gttgtcgatg agcttgtcaa ggttatgggt
     6301 cgtcacaagc ctgagaacat cgtcatcgag atggctcgtg agaaccagac tacccagaag
     6361 ggtcagaaga actcgaggga gcgcatgaag aggattgagg agggtatcaa ggagcttggt
     6421 tctcagatcc ttaaggagca ccctgtcgag aacacccagc tccagaacga gaagctctac
     6481 ctctactacc tccagaacgg tagggatatg tacgttgacc aggagctcga catcaacagg
     6541 ctttctgact acgacgtcga ccacattgtt cctcagtctt tccttaagga tgactccatc
     6601 gacaacaagg tcctcacgag gtccgacaag aacaggggta agtcggacaa cgtcccttcc
     6661 gaggaggttg tcaagaagat gaagaactac tggaggcagc ttctcaacgc taagctcatt
     6721 acccagagga agttcgacaa cctcacgaag gctgagaggg gtggcctttc cgagcttgac
     6781 aaggctggtt tcatcaagag gcagcttgtt gagacgaggc agattaccaa gcacgttgct
     6841 cagatcctcg attctaggat gaacaccaag tacgacgaga acgacaagct catccgcgag
     6901 gtcaaggtga tcaccctcaa gtccaagctc gtctccgact tccgcaagga cttccagttc
     6961 tacaaggtcc gcgagatcaa caactaccac cacgctcacg atgcttacct taacgctgtc
     7021 gttggtaccg ctcttatcaa gaagtaccct aagcttgagt ccgagttcgt ctacggtgac
     7081 tacaaggtct acgacgttcg taagatgatc gccaagtccg agcaggagat cggcaaggcc
     7141 accgccaagt acttcttcta ctccaacatc atgaacttct tcaagaccga gatcaccctc
     7201 gccaacggcg agatccgcaa gcgccctctt atcgagacga acggtgagac tggtgagatc
     7261 gtttgggaca agggtcgcga cttcgctact gttcgcaagg tcctttctat gcctcaggtt
     7321 aacatcgtca agaagaccga ggtccagacc ggtggcttct ccaaggagtc tatccttcca
     7381 aagagaaact cggacaagct catcgctagg aagaaggatt gggaccctaa gaagtacggt
     7441 ggtttcgact cccctactgt cgcctactcc gtcctcgtgg tcgccaaggt ggagaagggt
     7501 aagtcgaaga agctcaagtc cgtcaaggag ctcctcggca tcaccatcat ggagcgctcc
     7561 tccttcgaga agaacccgat cgacttcctc gaggccaagg gctacaagga ggtcaagaag
     7621 gacctcatca tcaagctccc caagtactct cttttcgagc tcgagaacgg tcgtaagagg
     7681 atgctggctt ccgctggtga gctccagaag ggtaacgagc ttgctcttcc ttccaagtac
     7741 gtgaacttcc tctacctcgc ctcccactac gagaagctca agggttcccc tgaggataac
     7801 gagcagaagc agctcttcgt ggagcagcac aagcactacc tcgacgagat catcgagcag
     7861 atctccgagt tctccaagcg cgtcatcctc gctgacgcta acctcgacaa ggtcctctcc
     7921 gcctacaaca agcaccgcga caagcccatc cgcgagcagg ccgagaacat catccacctc
     7981 ttcacgctca cgaacctcgg cgcccctgct gctttcaagt acttcgacac caccatcgac
     8041 aggaagcgtt acacgtccac caaggaggtt ctcgacgcta ctctcatcca ccagtccatc
     8101 accggtcttt acgagactcg tatcgacctt tcccagcttg gtggtgataa gcgtcctgct
     8161 gccaccaaaa aggccggaca ggctaagaaa aagaagtagg atcctcccga tcgttcaaac
     8221 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata
     8281 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt
     8341 atgaggtggg tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac
     8401 aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat
     8461 cgggagcacc ggtaaggcgc gccgtagtgc tcgagagacc tctgaagtgg ccgattcatt
     8521 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     8581 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     8641 tgttgtgtgg aattgtgagc ggataacaat ttcacacaag aaacagctat gaccatgatt
     8701 acgccaagct atttaggtga cactatagaa tactcaagct atgcatcaag ctcaatgggt
     8761 ctagtctgta gatacccatc acactggcga ccgctcgaac atcagtttaa ggtttacacc
     8821 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
     8881 ccggggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc
     8941 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
     9001 atggccagtg tgcctgtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
     9061 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggcctgaat
     9121 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgagcgt gggtctcgcg
     9181 gtatcattgg cgcgcctctc gagctagcgg ccgcatgcat cgatctccta catcgtataa
     9241 attagcctat acgaagttat tgcatctatg tcgggtgcgg agaaagaggt aatgaaatgg
     9301 cagtattaga tctgataact tcgtataatg tatgctatac gaagttatga ctgcaggtcg
     9361 acacccataa tagctgtttg ccaagcttgg cactggccgt cgtttttaca acgtcgtgac
     9421 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     9481 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     9541 ggcgaatgct agagcagctt gagcttggat cagattgtcg tttcccgcct tcagtttaaa
     9601 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag
     9661 aataacggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
     9721 gccaaccaca gggttcccct cgggatcaaa gtactttgat ccaacccctc cgctgctata
     9781 gtgcagtcgg cttctgacgt tcagtgcagc cgtcttctga aaacgacatg tcgcacaagt
     9841 cctaagttac gcgacaggct gccgccctgc ccttttcctg gcgttttctt gtcgcgtgtt
     9901 ttagtcgcat aaagtagaat acttgcgact agaaccggag acattacgcc atgaacaaga
     9961 gcgccgccgc tggcctgctg ggctatgccc gcgtcagcac cgacgaccag gacttgacca
    10021 accaacgggc cgaactgcac gcggccggct gcaccaagct gttttccgag aagatcaccg
    10081 gcaccaggcg cgaccgcccg gagctggcca ggatgcttga ccacctacgc cctggcgacg
    10141 ttgtgacagt gaccaggcta gaccgcctgg cccgcagcac ccgcgaccta ctggacattg
    10201 ccgagcgcat ccaggaggcc ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca
    10261 ccaccacgcc ggccggccgc atggtgttga ccgtgttcgc cggcattgcc gagttcgagc
    10321 gttccctaat catcgaccgc acccggagcg ggcgcgaggc cgccaaggcc cgaggcgtga
    10381 agtttggccc ccgccctacc ctcaccccgg cacagatcgc gcacgcccgc gagctgatcg
    10441 accaggaagg ccgcaccgtg aaagaggcgg ctgcactgct tggcgtgcat cgctcgaccc
    10501 tgtaccgcgc acttgagcgc agcgaggaag tgacgcccac cgaggccagg cggcgcggtg
    10561 ccttccgtga ggacgcattg accgaggccg acgccctggc ggccgccgag aatgaacgcc
    10621 aagaggaaca agcatgaaac cgcaccagga cggccaggac gaaccgtttt tcattaccga
    10681 agagatcgag gcggagatga tcgcggccgg gtacgtgttc gagccgcccg cgcacgtctc
    10741 aaccgtgcgg ctgcatgaaa tcctggccgg tttgtctgat gccaagctgg cggcctggcc
    10801 ggccagcttg gccgctgaag aaaccgagcg ccgccgtcta aaaaggtgat gtgtatttga
    10861 gtaaaacagc ttgcgtcatg cggtcgctgc gtatatgatg cgatgagtaa ataaacaaat
    10921 acgcaagggg aacgcatgaa ggttatcgct gtacttaacc agaaaggcgg gtcaggcaag
    10981 acgaccatcg caacccatct agcccgcgcc ctgcaactcg ccggggccga tgttctgtta
    11041 gtcgattccg atccccaggg cagtgcccgc gattgggcgg ccgtgcggga agatcaaccg
    11101 ctaaccgttg tcggcatcga ccgcccgacg attgaccgcg acgtgaaggc catcggccgg
    11161 cgcgacttcg tagtgatcga cggagcgccc caggcggcgg acttggctgt gtccgcgatc
    11221 aaggcagccg acttcgtgct gattccggtg cagccaagcc cttacgacat atgggccacc
    11281 gccgacctgg tggagctggt taagcagcgc attgaggtca cggatggaag gctacaagcg
    11341 gcctttgtcg tgtcgcgggc gatcaaaggc acgcgcatcg gcggtgaggt tgccgaggcg
    11401 ctggccgggt acgagctgcc cattcttgag tcccgtatca cgcagcgcgt gagctaccca
    11461 ggcactgccg ccgccggcac aaccgttctt gaatcagaac ccgagggcga cgctgcccgc
    11521 gaggtccagg cgctggccgc tgaaattaaa tcaaaactca tttgagttaa tgaggtaaag
    11581 agaaaatgag caaaagcaca aacacgctaa gtgccggccg tccgagcgca cgcagcagca
    11641 aggctgcaac gttggccagc ctggcagaca cgccagccat gaagcgggtc aactttcagt
    11701 tgccggcgga ggatcacacc aagctgaaga tgtacgcggt acgccaaggc aagaccatta
    11761 ccgagctgct atctgaatac atcgcgcagc taccagagta aatgagcaaa tgaataaatg
    11821 agtagatgaa ttttagcggc taaaggaggc ggcatggaaa atcaagaaca accaggcacc
    11881 gacgccgtgg aatgccccat gtgtggagga acgggcggtt ggccaggcgt aagcggctgg
    11941 gttgtctgcc ggccctgcaa tggcactgga acccccaagc ccgaggaatc ggcgtgacgg
    12001 tcgcaaacca tccggcccgg tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa
    12061 gttgaaggcc gcgcaggccg cccagcggca acgcatcgag gcagaagcac gccccggtga
    12121 atcgtggcaa gcggccgctg atcgaatccg caaagaatcc cggcaaccgc cggcagccgg
    12181 tgcgccgtcg attaggaagc cgcccaaggg cgacgagcaa ccagattttt tcgttccgat
    12241 gctctatgac gtgggcaccc gcgatagtcg cagcatcatg gacgtggccg ttttccgtct
    12301 gtcgaagcgt gaccgacgag ctggcgaggt gatccgctac gagcttccag acgggcacgt
    12361 agaggtttcc gcagggccgg ccggcatggc cagtgtgtgg gattacgacc tggtactgat
    12421 ggcggtttcc catctaaccg aatccatgaa ccgataccgg gaagggaagg gagacaagcc
    12481 cggccgcgtg ttccgtccac acgttgcgga cgtactcaag ttctgccggc gagccgatgg
    12541 cggaaagcag aaagacgacc tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc
    12601 catgcagcgt acgaagaagg ccaagaacgg ccgcctggtg acggtatccg agggtgaagc
    12661 cttgattagc cgctacaaga tcgtaaagag cgaaaccggg cggccggagt acatcgagat
    12721 cgagctagct gattggatgt accgcgagat cacagaaggc aagaacccgg acgtgctgac
    12781 ggttcacccc gattactttt tgatcgatcc cggcatcggc cgttttctct accgcctggc
    12841 acgccgcgcc gcaggcaagg cagaagccag atggttgttc aagacgatct acgaacgcag
    12901 tggcagcgcc ggagagttca agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa
    12961 tgacctgccg gagtacgatt tgaaggagga ggcggggcag gctggcccga tcctagtcat
    13021 gcgctaccgc aacctgatcg agggcgaagc atccgccggt tcctaatgta cggagcagat
    13081 gctagggcaa attgccctag caggggaaaa aggtcgaaaa ggtctgtttc ctgtggatag
    13141 cacgtacatt gggaacccaa agccgtacat tgggaaccgg aacccgtaca ttgggaaccc
    13201 aaagccgtac attgggaacc ggtcacacat gtaagtgact gatataaaag agaaaaaagg
    13261 cgatttttcc gcctaaaact ctttaaaact tattaaaact cttaaaaccc gcctggcctg
    13321 tgcataactg tctggccagc gcacagccga agagctgcaa aaagcgccta cccttcggtc
    13381 gctgcgctcc ctacgccccg ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa
    13441 aatggctggc ctacggccag gcaatctacc agggcgcgga caagccgcgc cgtcgccact
    13501 cgaccgccgg cgcccacatc aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa
    13561 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
    13621 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
    13681 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
    13741 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
    13801 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    13861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    13921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    13981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    14041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    14101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    14161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    14221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    14281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    14341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    14401 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
    14461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    14521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    14581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    14641 ttaagggatt ttggtcatgc attctaggta ctaaaacaat tcatccagta aaatataata
    14701 ttttattttc tcccaatcag gcttgatccc cagtaagtca aaaaatagct cgacatactg
    14761 ttcttccccg atatcctccc tgatcgaccg gacgcagaag gcaatgtcat accacttgtc
    14821 cgccctgccg cttctcccaa gatcaataaa gccacttact ttgccatctt tcacaaagat
    14881 gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt
    14941 taaaaaatca tacagctcgc gcggatcttt aaatggagtg tcttcttccc agttttcgca
    15001 atccacatcg gccagatcgt tattcagtaa gtaatccaat tcggctaagc ggctgtctaa
    15061 gctattcgta tagggacaat ccgatatgtc gatggagtga aagagcctga tgcactccgc
    15121 atacagctcg ataatctttt cagggctttg ttcatcttca tactcttccg agcaaaggac
    15181 gccatcggcc tcactcatga gcagattgct ccagccatca tgccgttcaa agtgcaggac
    15241 ctttggaaca ggcagctttc cttccagcca tagcatcatg tccttttccc gttccacatc
    15301 ataggtggtc cctttatacc ggctgtccgt catttttaaa tataggtttt cattttctcc
    15361 caccagctta tataccttag caggagacat tccttccgta tcttttacgc agcggtattt
    15421 ttcgatcagt tttttcaatt ccggtgatat tctcatttta gccatttatt atttccttcc
    15481 tcttttctac agtatttaaa gataccccaa gaagctaatt ataacaagac gaactccaat
    15541 tcactgttcc ttgcattcta aaaccttaaa taccagaaaa cagctttttc aaagttgttt
    15601 tcaaagttgg cgtataacat agtatcgacg gagccgattt tgaaaccgcg gtgatcacag
    15661 gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc tccgcgagat catccgtgtt
    15721 tcaaacccgg cagcttagtt gccgttcttc cgaatagcat cggtaacatg agcaaagtct
    15781 gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg cctgtatcga
    15841 gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctgg
//
