LOCUS       sequence-169495-       15945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of Cas9 in plants, cauliflower mosaic virus 35S promoter 
            (P35S), basta selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15945)
  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 15945)
  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-B
FEATURES             Location/Qualifiers
     source          1..15945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(281..957)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    1148..1169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1184..1189,1190..1207,1208..1214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1222..1238
                     /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     1246..1262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1637..1684)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(1685..5785)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(1685..5785)
                     /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             4591..4599
                     /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             complement(5810..5830)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(5979..6323)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    6836..6857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6872..6877,6878..6895,6896..6902)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6910..6926
                     /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        6968..6986
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6968..6985
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             7060..7362
                     /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
                     SRELYPVVHIGDGSWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     7241..7260
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    7572..7605
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(7649..7666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7649..7665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7658..7680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7868..7892
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             9193..9822
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10251..11324
                     /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"
     rep_origin      11390..11584
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11820..11842)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    11928..12068
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11983..12002
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12083..12100)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12254..12842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12334..12353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12929..13723)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    15347..15371
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    15449..15623
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(join(15630..15945,1..236))
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
ORIGIN
        1 gccttccagg ggcccgcgta ggcgatgccg gcgacctcgc cgtccacctc ggcgacgagc
       61 cagggatagc gctcccgcag acggacgagg tcgtccgtcc actcctgcgg ttcctgcggc
      121 tcggtacgga agttgaccgt gcttgtctcg atgtagtggt tgacgatggt gcagaccgcc
      181 ggcatgtccg cctcggtggc acggcggatg tcggccgggc gtcgttctgg gctcatggta
      241 gactcgagag agatagattt gtagagagag actggtgatt tcagcgtgtc ctctccaaat
      301 gaaatgaact tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat
      361 cccttacgtc agtggagata tcacatcaat ccacttgctt tgaagacgtg gttggaacgt
      421 cttctttttc cacgatgctc ttcgtgggtg ggggtccatc tttgggacca ctgtcggcag
      481 aggcatcttg aacgatagcc tttcctttat cgcaatgatg gcatttgtag gtgccacctt
      541 ccttttctac tgtccttttg atgaagtgac agatagctgg gcaatggaat ccgaggaggt
      601 ttcccgatat taccctttgt tgaaaagtct caatagccct ttggtcttct gagactgtat
      661 ctttgatatt cttggagtag acgagagtgt cgtgctccac catgttcaca tcaatccact
      721 tgctttgaag acgtggttgg aacgtcttct ttttccacga tgctcctcgt gggtgggggt
      781 ccatctttgg gaccactgtc ggcagaggca tcttgaacga tagcctttcc tttatcgcaa
      841 tgatggcatt tgtaggtgcc accttccttt tctactgtcc ttttgatgaa gtgacagata
      901 gctgggcaat ggaatccgag gaggtttccc gatattaccc tttgttgaaa agtctcaata
      961 gccctttggt cttctgagac tgtatctttg atattcttgg agtagacgag agtgtcgtgc
     1021 tccaccatgt tggcaagctg ctctagccaa tacgcaaacc gcctctcccc gcgcgttggc
     1081 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1141 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1201 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1261 acatgattac gaattcgagc tcggtaccgt aactataacg gtcctaaggt agcgaaggat
     1321 ccgctcgcta ccttaagaga ggatatccct ccatcctata atgtaggcta taggaactag
     1381 ggcaaggccg gccttatgcg gccgcgtcac tggattttgg ttttaggaat tagaaatttt
     1441 attgatagaa gtattttaca aatacaaata catactaagg gtttcttata tgctcaacac
     1501 atgagcgaaa ccctataaga accctaattc ccttatctgg gaactactca cacattattc
     1561 tggagaaaaa tagagagaga tagatttgta gagagagact ggtgattttt gcggactcta
     1621 gatcaggtgg atcctacttc tttttcttag cctgtccggc ctttttggtg gcagcaggac
     1681 gcttatcacc accaagctgg gaaaggtcga tacgagtctc gtaaagaccg gtgatggact
     1741 ggtggatgag agtagcgtcg agaacctcct tggtggacgt gtaacgcttc ctgtcgatgg
     1801 tggtgtcgaa gtacttgaaa gcagcagggg cgccgaggtt cgtgagcgtg aagaggtgga
     1861 tgatgttctc ggcctgctcg cggatgggct tgtcgcggtg cttgttgtag gcggagagga
     1921 ccttgtcgag gttagcgtca gcgaggatga cgcgcttgga gaactcggag atctgctcga
     1981 tgatctcgtc gaggtagtgc ttgtgctgct ccacgaagag ctgcttctgc tcgttatcct
     2041 caggggaacc cttgagcttc tcgtagtggg aggcgaggta gaggaagttc acgtacttgg
     2101 aaggaagagc aagctcgtta cccttctgga gctcaccagc ggaagccagc atcctcttac
     2161 gaccgttctc gagctcgaaa agagagtact tggggagctt gatgatgagg tccttcttga
     2221 cctccttgta gcccttggcc tcgaggaagt cgatcgggtt cttctcgaag gaggagcgct
     2281 ccatgatggt gatgccgagg agctccttga cggacttgag cttcttcgac ttacccttct
     2341 ccaccttggc gaccacgagg acggagtagg cgacagtagg ggagtcgaaa ccaccgtact
     2401 tcttagggtc ccaatccttc ttcctagcga tgagcttgtc cgagtttctc tttggaagga
     2461 tagactcctt ggagaagcca ccggtctgga cctcggtctt cttgacgatg ttaacctgag
     2521 gcatagaaag gaccttgcga acagtagcga agtcgcgacc cttgtcccaa acgatctcac
     2581 cagtctcacc gttcgtctcg ataagagggc gcttgcggat ctcgccgttg gcgagggtga
     2641 tctcggtctt gaagaagttc atgatgttgg agtagaagaa gtacttggcg gtggccttgc
     2701 cgatctcctg ctcggacttg gcgatcatct tacgaacgtc gtagaccttg tagtcaccgt
     2761 agacgaactc ggactcaagc ttagggtact tcttgataag agcggtacca acgacagcgt
     2821 taaggtaagc atcgtgagcg tggtggtagt tgttgatctc gcggaccttg tagaactgga
     2881 agtccttgcg gaagtcggag acgagcttgg acttgagggt gatcaccttg acctcgcgga
     2941 tgagcttgtc gttctcgtcg tacttggtgt tcatcctaga atcgaggatc tgagcaacgt
     3001 gcttggtaat ctgcctcgtc tcaacaagct gcctcttgat gaaaccagcc ttgtcaagct
     3061 cggaaaggcc acccctctca gccttcgtga ggttgtcgaa cttcctctgg gtaatgagct
     3121 tagcgttgag aagctgcctc cagtagttct tcatcttctt gacaacctcc tcggaaggga
     3181 cgttgtccga cttacccctg ttcttgtcgg acctcgtgag gaccttgttg tcgatggagt
     3241 catccttaag gaaagactga ggaacaatgt ggtcgacgtc gtagtcagaa agcctgttga
     3301 tgtcgagctc ctggtcaacg tacatatccc taccgttctg gaggtagtag aggtagagct
     3361 tctcgttctg gagctgggtg ttctcgacag ggtgctcctt aaggatctga gaaccaagct
     3421 ccttgatacc ctcctcaatc ctcttcatgc gctccctcga gttcttctga cccttctggg
     3481 tagtctggtt ctcacgagcc atctcgatga cgatgttctc aggcttgtga cgacccataa
     3541 ccttgacaag ctcatcgaca accttaacag tctgaaggat gcccttcttg atagcagggg
     3601 aaccagcaag gttagcaatg tgctcgtgga gagagtcgcc ctgaccggac acctgagcct
     3661 tctgaatatc ctccttgaag gtaagagagt catcgtggat gagctgcatg aagttcctgt
     3721 tagcgaaacc atcagacttg aggaagtcga ggatagtctt gccgctctgc ttatccctga
     3781 taccgttgat gagcttgcgg gagagcctac cccaaccggt gtaacggcga cgcttgagct
     3841 gcttcataac cttgtcatcg aagagatgag cgtaagtctt gagcctctcc tcgatcatct
     3901 ccctatcctc gaagagagta agagtgagga cgatgtcctc gaggatgtcc tcgttctcct
     3961 cgttgtcgag gaagtccttg tccttgatga tcttgaggag atcgtggtag gtaccgagag
     4021 aagcgttgaa acggtcctca acgccgctga tctcgacgga gtcgaagcac tcgatcttct
     4081 tgaagtagtc ctccttgagc tgcttgacgg tgaccttgcg gttggtcttg aagaggaggt
     4141 caacgatagc cttcttctgc tcgccggaga ggaaggcagg cttgcgcata ccctcggtga
     4201 cgtacttgac cttggtgagc tcgttgtaga ccgtgaagta ctcgtagagg agggagtgct
     4261 tggggaggac cttctcgttg gggaggttct tgtcgaagtt ggtcatgcgc tcgatgaagg
     4321 actgggcgga agcacccttg tcaacaacct cctcgaagtt ccaaggggtg atagtctcct
     4381 cggacttcct agtcatccaa gcgaaacggg agttaccacg agcaagagga ccaacgtagt
     4441 aaggaatacg gaaagtaagg atcttctcga tcttctcacg gttgtccttg aggaaagggt
     4501 agaagtcctc ctgcctacga aggatggcgt gaagctcacc aaggtggatc tggtgaggga
     4561 tagagccgtt atcgaaagtc ctctgcttcc taaggaggtc ctcacggtta agcttaacga
     4621 gaagctcctc agtaccatcc atcttctcaa ggattggctt gatgaacttg tagaactcct
     4681 cctggctagc tccaccgtca atgtaaccgg cgtagccgtt cttggactgg tcgaagaaga
     4741 tctccttgta cttctcgggg agctgctgac gaacaagagc cttgaggagg gtgaggtcct
     4801 ggtggtgctc gtcgtagcgc ttgatcatgg aagcagaaag aggagcctta gtgatctcag
     4861 tgttaaccct aaggatatcc gaaaggagga tagcatcgga gaggttctta gcagcaagga
     4921 agagatcagc gtactgatct ccaatctgag cgaggaggtt gtcgagatca tcgtcgtagg
     4981 tatccttgga gagctgaagc ttagcgtcct cagcgaggtc gaagttggac ttgaagttag
     5041 gggtcagacc gagggagaga gcgatgaggt taccgaaaag accgttcttc ttctcaccag
     5101 ggagctgggc gatgaggttc tcgagacgcc tggacttgga gagcctagcg gaaaggatcg
     5161 ccttagcgtc gacaccggaa gcgttgatag ggttctcctc gaagagctgg ttgtaggtct
     5221 ggacgagctg gatgaagagc ttgtccacgt cggagttatc agggttaagg tcaccctcga
     5281 taaggaagtg accacggaac ttgatcatgt gagcgagagc aaggtagatg agacgaagat
     5341 cagccttatc agtagagtca acaagcttct tacgaaggtg gtagatagtg gggtacttct
     5401 cgtggtaggc gacctcgtcg acgatgttgc cgaagatggg gtggcgctcg tgcttcttgt
     5461 cctcctccac gaggaaggac tcctcgaggc ggtggaagaa ggagtcgtcg accttcgcca
     5521 tctcgttgga gaagatctcc tggaggtagc agatgcggtt cttgcggcgg gtgtagcggc
     5581 ggcgggcggt gcgcttgagg cgggtcgcct ccgccgtctc gccggagtcg aagaggaggg
     5641 cgccgatgag gttcttcttg atggagtggc ggtcggtgtt gcccaggacc ttgaacttct
     5701 tggacgggac cttgtactcg tcggtgatca ccgcccagcc gacgctgttg gtgccgatgt
     5761 cgaggccgat ggagtacttc ttgtcagccg caggcacccc gtgaatacca accttccgct
     5821 tcttcttagg agccatggct atcgttcgta aatggtgaaa attttcagaa aattgctttt
     5881 gctttaaaag aaatgattta aattgctgca atagaagtag aatgcttgat tgcttgagat
     5941 tcgtttgttt tgtatatgtt gtgttgagaa ctctcgacgt cctctccaaa tgaaatgaac
     6001 ttccttatat agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt
     6061 cagtggagat atcacatcaa tccacttgct ttgaagacgt ggttggaacg tcttcttttt
     6121 ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca gaggcatctt
     6181 caacgatggc ctttccttta tcgcaatgat ggcatttgta ggagccacct tccttttcca
     6241 ctatcttcac aataaagtga cagatagctg ggcaatggaa tccgaggagg tttccggata
     6301 ttaccctttg ttgaaaagtc tcaattgccc tttggtcttc tgagactgta tctttgatat
     6361 ttttggagta gacaagtgtg tcgtgctcca ccatgttatc acatcaatcc acttgctttg
     6421 aagacgtggt tggaacgtct tctttttcca cgatgctcct cgtgggtggg ggtccatctt
     6481 tgggaccact gtcggcagag gcatcttcaa cgatggcctt tcctttatcg caatgatggc
     6541 atttgtagga gccaccttcc ttttccacta tcttcacaat aaagtgacag atagctgggc
     6601 aatggaatcc gaggaggttt ccggatatta ccctttgttg aaaagtctca attgcccttt
     6661 ggtcttctga gactgtatct ttgatatttt tggagtagac aagtgtgtcg tgctccacat
     6721 gttgaccggt aaggcgcgcc gtagtgctcg agagacctct gaagtggccg attcattaat
     6781 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6841 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6901 tgtgtggaat tgtgagcgga taacaatttc acacaagaaa cagctatgac catgattacg
     6961 ccaagctatt taggtgacac tatagaatac tcaagctatg catcaagctc aatgggtcta
     7021 gtctgtagat acccatcaca ctggcgaccg ctcgaacatc agtttaaggt ttacacctat
     7081 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccg
     7141 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     7201 gaactttacc cggtggtgca tatcggggat ggaagctggc gcatgatgac caccgatatg
     7261 gccagtgtgc ctgtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     7321 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg cctgaatggc
     7381 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgagcgtggg tctcgcggta
     7441 tcattggcgc gcctctcgag ctagcggccg catgcatcga tctcctacat cgtataaatt
     7501 agcctatacg aagttattgc atctatgtcg ggtgcggaga aagaggtaat gaaatggcag
     7561 tattagatct gataacttcg tataatgtat gctatacgaa gttatgactg caggtcgaca
     7621 cccataatag ctgtttgcca agcttggcac tggccgtcgt tttacaacgt cgtgactggg
     7681 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     7741 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     7801 aatgctagag cagcttgagc ttggatcaga ttgtcgtttc ccgccttcag tttaaactat
     7861 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     7921 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc
     7981 aaccacaggg ttcccctcgg gatcaaagta ctttgatcca acccctccgc tgctatagtg
     8041 cagtcggctt ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct
     8101 aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta
     8161 gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg
     8221 ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc
     8281 aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca
     8341 ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct ggcgacgttg
     8401 tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg gacattgccg
     8461 agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg gccgacacca
     8521 ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt
     8581 ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt
     8641 ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag ctgatcgacc
     8701 aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt
     8761 accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct
     8821 tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat gaacgccaag
     8881 aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca ttaccgaaga
     8941 gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc acgtctcaac
     9001 cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg cctggccggc
     9061 cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta
     9121 aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata aacaaatacg
     9181 caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc aggcaagacg
     9241 accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt tctgttagtc
     9301 gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta
     9361 accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat cggccggcgc
     9421 gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc cgcgatcaag
     9481 gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg ggccaccgcc
     9541 gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct acaagcggcc
     9601 tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg
     9661 gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc
     9721 actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag
     9781 gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga ggtaaagaga
     9841 aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg
     9901 ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac tttcagttgc
     9961 cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag accattaccg
    10021 agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga ataaatgagt
    10081 agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac
    10141 gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag cggctgggtt
    10201 gcctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgagcggtc
    10261 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
    10321 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
    10381 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
    10441 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
    10501 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
    10561 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
    10621 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
    10681 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
    10741 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
    10801 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
    10861 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
    10921 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
    10981 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
    11041 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
    11101 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
    11161 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
    11221 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
    11281 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
    11341 tagggcaaat tgccctagca ggggaaaaag gtcgaaaagt tctctttcct gtggatagca
    11401 cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa
    11461 agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg
    11521 atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg
    11581 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
    11641 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
    11701 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
    11761 accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac
    11821 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
    11881 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
    11941 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
    12001 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
    12061 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    12121 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    12181 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    12241 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    12301 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    12361 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    12421 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    12481 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    12541 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    12601 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    12661 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
    12721 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    12781 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    12841 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    12901 aagggatttt ggtcatgcat tctaggtact aaaacaattc atccagtaaa atataatatt
    12961 ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt
    13021 cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac cacttgtccg
    13081 ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt
    13141 tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta
    13201 aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag ttttcgcaat
    13261 ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc
    13321 tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat
    13381 acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc
    13441 catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct
    13501 ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat
    13561 aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca
    13621 ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt
    13681 cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat ttccttcctc
    13741 ttttctacag tatttaaaga taccccaaga agctaaggaa ggtgcgaaca agtccctgat
    13801 atgagatcat gtttgtcatc tggagccata gaacagggtt catcatgagt catcaactta
    13861 ccttcgccga cagtgaattc agcagtaagc gccgtcagac cagaaaagag attttcttgt
    13921 cccgcatgga gcagattctg ccatggcaaa acatggtgga agtcatcgag ccgttttacc
    13981 ccaaggctgg taatggccgg cgaccttatc cgctggaaac catgctacgc attcactgca
    14041 tgcagcattg gtacaacctg agcgatggcg cgatggaaga tgctctgtac gaaatcgcct
    14101 ccatgcgtct gtttgcccgg ttatccctgg atagcgcctt gccggaccgc accaccatca
    14161 tgaatttccg ccacctgctg gagcagcatc aactggcccg ccaattgttc aagaccatca
    14221 atcgctggct ggccgaagca ggcgtcatga tgactcaagg caccttggtc gatgccacca
    14281 tcattgaggc acccagctcg accaagaaca aagagcagca acgcgatccg gagatgcatc
    14341 agaccaagaa aggcaatcag tggcactttg gcatgaaggc ccacattggt gtcgatgcca
    14401 agagtggcct gacccacagc ctggtcacca ccgcggccaa cgagcatgac ctcaatcagc
    14461 tgggtaatct gctgcatgga gaggagcaat ttgtctcagc cgatgccggc taccaagggg
    14521 cgccacagcg cgaggagctg gccgaggtgg atgtggactg gctgatcgcc gagcgccccg
    14581 gcaaggtaag aaccttgaaa cagcatccac gcaagaacaa aacggccatc aacatcgaat
    14641 acatgaaagc cagcatccgg gccagggtgg agcacccatt tcgcatcatc aagcgacagt
    14701 tcggcttcgt gaaagccaga tacaaggggt tgctgaaaaa cgataaccaa ctggcgatgt
    14761 tattcacgct ggccaacctg tttcgggcgg accaaatgat acgtcagtgg gagagatctc
    14821 actaaaaact ggggataacg ccttaaatgg cgaagaaacg gtctaaatag gctgattcaa
    14881 ggcatttacg ggagaaaaaa tcggctcaaa catgaagaaa tgaaatgact gagtcagccg
    14941 agaagaattt ccccgcttat tcgcaccttc cctaattata acaagacgaa ctccaattca
    15001 ctgttccttg cattctaaaa ccttaaatac cagaaaacag ctttttcaaa gttgttttca
    15061 aagttggcgt ataacatagt atcgacggag ccgattttga aaccgcggtg atcacaggca
    15121 gcaacgctct gtcatcgtta caatcaacat gctaccctcc gcgagatcat ccgtgtttca
    15181 aacccggcag cttagttgcc gttcttccga atagcatcgg taacatgagc aaagtctgcc
    15241 gccttacaac ggctctcccg ctgacgccgt cccggactga tgggctgcct gtatcgagtg
    15301 gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt
    15361 gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg tttttaatgt
    15421 actgaattaa cgccgaatta attcggggga tctggatttt agtactggat tttggtttta
    15481 ggaattagaa attttattga tagaagtatt ttacaaatac aaatacatac taagggtttc
    15541 ttatatgctc aacacatgag cgaaacccta taggaaccct aattccctta tctgggaact
    15601 actcacacat tattatggag aaactcgagt caaatctcgg tgacgggcag gaccggacgg
    15661 ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc
    15721 ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc
    15781 acgctcgggt cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc
    15841 agggacttca gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg
    15901 gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt tgcgt
//
