LOCUS       sequence-201332-       16203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Cas9 in plants, ZmUbi promoter, Kanamycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16203)
  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 16203)
  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-N
FEATURES             Location/Qualifiers
     source          1..16203
                     /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(334..1131)
                     /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(445..464)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1055..1074
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1194..1871)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    2062..2083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2098..2103,2104..2121,2122..2128)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2136..2152
                     /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     2160..2176
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4317..4337
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4362..8462
                     /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             4362..8462
                     /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             5548..5556
                     /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             8463..8510
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8523..8775
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8893..8914
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8929..8934,8935..8952,8953..8959)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8967..8983
                     /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        9025..9043
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     9025..9042
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             9117..9419
                     /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     9298..9317
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    9629..9662
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9926..9950
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             11250..11879
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             12313..13380
                     /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      13446..13640
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(13876..13898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    13984..14124
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     14039..14058
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(14139..14156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14310..14898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14390..14409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14985..15779)
                     /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 gagcttgtcg atcgactcta
      301 gctagaggat cgatccgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
      361 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
      421 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
      481 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
      541 gatattcggc aagcaggcat cgccatgtgt cacgacgaga tcctcgccgt cgggcatgcg
      601 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
      661 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
      721 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
      781 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
      841 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
      901 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgga gttcattcag
      961 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccgaaa
     1021 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     1081 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcccca tggtcgatcg
     1141 acagatctgc gaaagctcga gagagataga tttgtagaga gagactggtg atttcagcgt
     1201 gtcctctcca aatgaaatga acttccttat atagaggaag gtcttgcgaa ggatagtggg
     1261 attgtgcgtc atcccttacg tcagtggaga tatcacatca atccacttgc tttgaagacg
     1321 tggttggaac gtcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac
     1381 cactgtcggc agaggcatct tgaacgatag cctttccttt atcgcaatga tggcatttgt
     1441 aggtgccacc ttccttttct actgtccttt tgatgaagtg acagatagct gggcaatgga
     1501 atccgaggag gtttcccgat attacccttt gttgaaaagt ctcaatagcc ctttggtctt
     1561 ctgagactgt atctttgata ttcttggagt agacgagagt gtcgtgctcc accatgttat
     1621 cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc
     1681 tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct
     1741 ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga
     1801 tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt
     1861 gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga
     1921 cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgcctct
     1981 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2041 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     2101 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     2161 aggaaacagc tatgaccatg attacgaatt cgagctcggt accaactata acggtcctaa
     2221 ggtagcgaag gatccgctcg ctaccttaag agaggatatc cctccatcct ataatgtagg
     2281 ctataggaac tagggcaagg ccggccatgc ggccgcaagc tgggtgcagc gtgacccggt
     2341 cgtgcccctc tctagagata atgagcattg catgtctaag ttataaaaaa ttaccacata
     2401 ttttttttgt cacacttgtt tgaagtgcag tttatctatc tttatacata tatttaaact
     2461 ttactctacg aataatataa tctatagtac tacaataata tcagtgtttt agagaatcat
     2521 ataaatgaac agttagacat ggtctaaagg acaattgagt attttgacaa caggactcta
     2581 cagttttatc tttttagtgt gcatgtgttc tccttttttt ttgcaaatag cttcacctat
     2641 ataatacttc atccatttta ttagtacatc catttagggt ttagggttaa tggtttttat
     2701 agactaattt ttttagtaca tctattttat tctattttag cctctaaatt aagaaaacta
     2761 aaactctatt ttagtttttt tatttaataa tttagatata aaatagaata aaataaagtg
     2821 actaaaaatt aaacaaatac cctttaagaa attaaaaaaa ctaaggaaac atttttcttg
     2881 tttcgagtag ataatgccag cctgttaaac gccgtcgacg agtctaacgg acaccaacca
     2941 gcgaaccagc agcgtcgcgt cgggccaagc gaagcagacg gcacggcatc tctgtcgctg
     3001 cctctggacc cctctcgaga gttccgctcc accgttggac ttgctccgct gtcggcatcc
     3061 agaaattgcg tggcggagcg gcagacgtga gccggcacgg caggcggcct cctcctcctc
     3121 tcacggcacg gcagctacgg gggattcctt tcccaccgct ccttcgcttt cccttcctcg
     3181 cccgccgtaa taaatagaca ccccctccac accctctttc cccaacctcg tgttgttcgg
     3241 agcgcacaca cacacaacca gatctccccc aaatccaccc gtcggcacct ccgcttcaag
     3301 gtacgccgct cgtcctcccc ccccccccct ctctaccttc tctagatcgg cgttccggtc
     3361 catggttagg gcccggtagt tctacttctg ttcatgtttg tgttagatcc gtgtttgtgt
     3421 tagatccgtg ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac acgttctgat
     3481 tgctaacttg ccagtgtttc tctttgggga atcctgggat ggctctagcc gttccgcaga
     3541 cgggatcgat ttcatgattt tttttgtttc gttgcatagg gtttggtttg cccttttcct
     3601 ttatttcaat atatgccgtg cacttgtttg tcgggtcatc ttttcatgct tttttttgtc
     3661 ttggttgtga tgatgtggtc tggttgggcg gtcgttctag atcggagtag aattctgttt
     3721 caaactacct ggtggattta ttaattttgg atctgtatgt gtgtgccata catattcata
     3781 gttacgaatt gaagatgatg gatggaaata tcgatctagg ataggtatac atgttgatgc
     3841 gggttttact gatgcatata cagagatgct ttttgttcgc ttggttgtga tgatgtggtg
     3901 tggttgggcg gtcgttcatt cgttctagat cggagtagaa tactgtttca aactacctgg
     3961 tgtatttatt aattttggaa ctgtatgtgt gtgtcataca tcttcatagt tacgagttta
     4021 agatggatgg aaatatcgat ctaggatagg tatacatgtt gatgtgggtt ttactgatgc
     4081 atatacatga tggcatatgc agcatctatt catatgctct aaccttgagt acctatctat
     4141 tataataaac aagtatgttt tataattatt ttgatcttga tatacttgga tgatggcata
     4201 tgcagcagct atatgtggat ttttttagcc ctgccttcat acgctattta tttgcttggt
     4261 actgtttctt ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag atggctccta
     4321 agaagaagcg gaaggttggt attcacgggg tgcctgcggc tgacaagaag tactccatcg
     4381 gcctcgacat cggcaccaac agcgtcggct gggcggtgat caccgacgag tacaaggtcc
     4441 cgtccaagaa gttcaaggtc ctgggcaaca ccgaccgcca ctccatcaag aagaacctca
     4501 tcggcgccct cctcttcgac tccggcgaga cggcggaggc gacccgcctc aagcgcaccg
     4561 cccgccgccg ctacacccgc cgcaagaacc gcatctgcta cctccaggag atcttctcca
     4621 acgagatggc gaaggtcgac gactccttct tccaccgcct cgaggagtcc ttcctcgtgg
     4681 aggaggacaa gaagcacgag cgccacccca tcttcggcaa catcgtcgac gaggtcgcct
     4741 accacgagaa gtaccccact atctaccacc ttcgtaagaa gcttgttgac tctactgata
     4801 aggctgatct tcgtctcatc taccttgctc tcgctcacat gatcaagttc cgtggtcact
     4861 tccttatcga gggtgacctt aaccctgata actccgacgt ggacaagctc ttcatccagc
     4921 tcgtccagac ctacaaccag ctcttcgagg agaaccctat caacgcttcc ggtgtcgacg
     4981 ctaaggcgat cctttccgct aggctctcca agtccaggcg tctcgagaac ctcatcgccc
     5041 agctccctgg tgagaagaag aacggtcttt tcggtaacct catcgctctc tccctcggtc
     5101 tgacccctaa cttcaagtcc aacttcgacc tcgctgagga cgctaagctt cagctctcca
     5161 aggataccta cgacgatgat ctcgacaacc tcctcgctca gattggagat cagtacgctg
     5221 atctcttcct tgctgctaag aacctctccg atgctatcct cctttcggat atccttaggg
     5281 ttaacactga gatcactaag gctcctcttt ctgcttccat gatcaagcgc tacgacgagc
     5341 accaccagga cctcaccctc ctcaaggctc ttgttcgtca gcagctcccc gagaagtaca
     5401 aggagatctt cttcgaccag tccaagaacg gctacgccgg ttacattgac ggtggagcta
     5461 gccaggagga gttctacaag ttcatcaagc caatccttga gaagatggat ggtactgagg
     5521 agcttctcgt taagcttaac cgtgaggacc tccttaggaa gcagaggact ttcgataacg
     5581 gctctatccc tcaccagatc caccttggtg agcttcacgc catccttcgt aggcaggagg
     5641 acttctaccc tttcctcaag gacaaccgtg agaagatcga gaagatcctt actttccgta
     5701 ttccttacta cgttggtcct cttgctcgtg gtaactcccg tttcgcttgg atgactagga
     5761 agtccgagga gactatcacc ccttggaact tcgaggaggt tgttgacaag ggtgcttccg
     5821 cccagtcctt catcgagcgc atgaccaact tcgacaagaa cctccccaac gagaaggtcc
     5881 tccccaagca ctccctcctc tacgagtact tcacggtcta caacgagctc accaaggtca
     5941 agtacgtcac cgagggtatg cgcaagcctg ccttcctctc cggcgagcag aagaaggcta
     6001 tcgttgacct cctcttcaag accaaccgca aggtcaccgt caagcagctc aaggaggact
     6061 acttcaagaa gatcgagtgc ttcgactccg tcgagatcag cggcgttgag gaccgtttca
     6121 acgcttctct cggtacctac cacgatctcc tcaagatcat caaggacaag gacttcctcg
     6181 acaacgagga gaacgaggac atcctcgagg acatcgtcct cactcttact ctcttcgagg
     6241 atagggagat gatcgaggag aggctcaaga cttacgctca tctcttcgat gacaaggtta
     6301 tgaagcagct caagcgtcgc cgttacaccg gttggggtag gctctcccgc aagctcatca
     6361 acggtatcag ggataagcag agcggcaaga ctatcctcga cttcctcaag tctgatggtt
     6421 tcgctaacag gaacttcatg cagctcatcc acgatgactc tcttaccttc aaggaggata
     6481 ttcagaaggc tcaggtgtcc ggtcagggcg actctctcca cgagcacatt gctaaccttg
     6541 ctggttcccc tgctatcaag aagggcatcc ttcagactgt taaggttgtc gatgagcttg
     6601 tcaaggttat gggtcgtcac aagcctgaga acatcgtcat cgagatggct cgtgagaacc
     6661 agactaccca gaagggtcag aagaactcga gggagcgcat gaagaggatt gaggagggta
     6721 tcaaggagct tggttctcag atccttaagg agcaccctgt cgagaacacc cagctccaga
     6781 acgagaagct ctacctctac tacctccaga acggtaggga tatgtacgtt gaccaggagc
     6841 tcgacatcaa caggctttct gactacgacg tcgaccacat tgttcctcag tctttcctta
     6901 aggatgactc catcgacaac aaggtcctca cgaggtccga caagaacagg ggtaagtcgg
     6961 acaacgtccc ttccgaggag gttgtcaaga agatgaagaa ctactggagg cagcttctca
     7021 acgctaagct cattacccag aggaagttcg acaacctcac gaaggctgag aggggtggcc
     7081 tttccgagct tgacaaggct ggtttcatca agaggcagct tgttgagacg aggcagatta
     7141 ccaagcacgt tgctcagatc ctcgattcta ggatgaacac caagtacgac gagaacgaca
     7201 agctcatccg cgaggtcaag gtgatcaccc tcaagtccaa gctcgtctcc gacttccgca
     7261 aggacttcca gttctacaag gtccgcgaga tcaacaacta ccaccacgct cacgatgctt
     7321 accttaacgc tgtcgttggt accgctctta tcaagaagta ccctaagctt gagtccgagt
     7381 tcgtctacgg tgactacaag gtctacgacg ttcgtaagat gatcgccaag tccgagcagg
     7441 agatcggcaa ggccaccgcc aagtacttct tctactccaa catcatgaac ttcttcaaga
     7501 ccgagatcac cctcgccaac ggcgagatcc gcaagcgccc tcttatcgag acgaacggtg
     7561 agactggtga gatcgtttgg gacaagggtc gcgacttcgc tactgttcgc aaggtccttt
     7621 ctatgcctca ggttaacatc gtcaagaaga ccgaggtcca gaccggtggc ttctccaagg
     7681 agtctatcct tccaaagaga aactcggaca agctcatcgc taggaagaag gattgggacc
     7741 ctaagaagta cggtggtttc gactccccta ctgtcgccta ctccgtcctc gtggtcgcca
     7801 aggtggagaa gggtaagtcg aagaagctca agtccgtcaa ggagctcctc ggcatcacca
     7861 tcatggagcg ctcctccttc gagaagaacc cgatcgactt cctcgaggcc aagggctaca
     7921 aggaggtcaa gaaggacctc atcatcaagc tccccaagta ctctcttttc gagctcgaga
     7981 acggtcgtaa gaggatgctg gcttccgctg gtgagctcca gaagggtaac gagcttgctc
     8041 ttccttccaa gtacgtgaac ttcctctacc tcgcctccca ctacgagaag ctcaagggtt
     8101 cccctgagga taacgagcag aagcagctct tcgtggagca gcacaagcac tacctcgacg
     8161 agatcatcga gcagatctcc gagttctcca agcgcgtcat cctcgctgac gctaacctcg
     8221 acaaggtcct ctccgcctac aacaagcacc gcgacaagcc catccgcgag caggccgaga
     8281 acatcatcca cctcttcacg ctcacgaacc tcggcgcccc tgctgctttc aagtacttcg
     8341 acaccaccat cgacaggaag cgttacacgt ccaccaagga ggttctcgac gctactctca
     8401 tccaccagtc catcaccggt ctttacgaga ctcgtatcga cctttcccag cttggtggtg
     8461 ataagcgtcc tgctgccacc aaaaaggccg gacaggctaa gaaaaagaag taggatcctc
     8521 ccgatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg
     8581 cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat
     8641 gcatgacgtt atttatgagg tgggttttta tgattagagt cccgcaatta tacatttaat
     8701 acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat
     8761 ctatgttact agatcgggag caccggtaag gcgcgccgta gtgctcgaga gacctctgaa
     8821 gtggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     8881 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     8941 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caagaaacag
     9001 ctatgaccat gattacgcca agctatttag gtgacactat agaatactca agctatgcat
     9061 caagctcaat gggtctagtc tgtagatacc catcacactg gcgaccgctc gaacatcagt
     9121 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     9181 atattattga cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     9241 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     9301 tgatgaccac cgatatggcc agtgtgcctg tctccgttat cggggaagaa gtggctgatc
     9361 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     9421 tgtcaggcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtga
     9481 gcgtgggtct cgcggtatca ttggcgcgcc tctcgagcta gcggccgcat gcatcgatct
     9541 cctacatcgt ataaattagc ctatacgaag ttattgcatc tatgtcgggt gcggagaaag
     9601 aggtaatgaa atggcagtat tagatctgat aacttcgtat aatgtatgct atacgaagtt
     9661 atgactgcag gtcgacaccc ataatagctg tttgccaagc ttggcactgg ccgtcgtttt
     9721 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     9781 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     9841 tgcgcagcct gaatggcgaa tgctagagca gcttgagctt ggatcagatt gtcgtttccc
     9901 gccttcagtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
     9961 agagcgttta ttagaataac ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
    10021 atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt tgatccaacc
    10081 cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt ctgaaaacga
    10141 catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt cctggcgttt
    10201 tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc ggagacatta
    10261 cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca gcaccgacga
    10321 ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca agctgttttc
    10381 cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc ttgaccacct
    10441 acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca gcacccgcga
    10501 cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta gcctggcaga
    10561 gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt tcgccggcat
    10621 tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg aggccgccaa
    10681 ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga tcgcgcacgc
    10741 ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac tgcttggcgt
    10801 gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc ccaccgaggc
    10861 caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc tggcggccgc
    10921 cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca ggacgaaccg
    10981 tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt gttcgagccg
    11041 cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc tgatgccaag
    11101 ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg tctaaaaagg
    11161 tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat gatgcgatga
    11221 gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt aaccagaaag
    11281 gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa ctcgccgggg
    11341 ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg gcggccgtgc
    11401 gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac cgcgacgtga
    11461 aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg gcggacttgg
    11521 ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca agcccttacg
    11581 acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag gtcacggatg
    11641 gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc atcggcggtg
    11701 aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt atcacgcagc
    11761 gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca gaacccgagg
    11821 gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa ctcatttgag
    11881 ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg gccgtccgag
    11941 cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag ccatgaagcg
    12001 ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg cggtacgcca
    12061 aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag agtaaatgag
    12121 caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg gaaaatcaag
    12181 aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc ggttggccag
    12241 gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc aagcccgagg
    12301 aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat
    12361 gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa
    12421 gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa
    12481 ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat
    12541 tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg
    12601 gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt
    12661 ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac
    12721 gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg
    12781 aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc
    12841 cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac
    12901 accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta
    12961 tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg
    13021 gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac
    13081 ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt
    13141 ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg
    13201 atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag
    13261 ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc
    13321 ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa
    13381 tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggtctg
    13441 tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg
    13501 tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata
    13561 aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa
    13621 acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg
    13681 cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc
    13741 gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc
    13801 gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg
    13861 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
    13921 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
    13981 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
    14041 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
    14101 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
    14161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
    14221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
    14281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
    14341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
    14401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
    14461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
    14521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
    14581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
    14641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
    14701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
    14761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
    14821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
    14881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
    14941 aacgaaaact cacgttaagg gattttggtc atgcattcta ggtactaaaa caattcatcc
    15001 agtaaaatat aatattttat tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat
    15061 agctcgacat actgttcttc cccgatatcc tccctgatcg accggacgca gaaggcaatg
    15121 tcataccact tgtccgccct gccgcttctc ccaagatcaa taaagccact tactttgcca
    15181 tctttcacaa agatgttgct gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg
    15241 ggcttttccg tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg agtgtcttct
    15301 tcccagtttt cgcaatccac atcggccaga tcgttattca gtaagtaatc caattcggct
    15361 aagcggctgt ctaagctatt cgtataggga caatccgata tgtcgatgga gtgaaagagc
    15421 ctgatgcact ccgcatacag ctcgataatc ttttcagggc tttgttcatc ttcatactct
    15481 tccgagcaaa ggacgccatc ggcctcactc atgagcagat tgctccagcc atcatgccgt
    15541 tcaaagtgca ggacctttgg aacaggcagc tttccttcca gccatagcat catgtccttt
    15601 tcccgttcca catcataggt ggtcccttta taccggctgt ccgtcatttt taaatatagg
    15661 ttttcatttt ctcccaccag cttatatacc ttagcaggag acattccttc cgtatctttt
    15721 acgcagcggt atttttcgat cagttttttc aattccggtg atattctcat tttagccatt
    15781 tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca
    15841 agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt
    15901 tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac
    15961 cgcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct accctccgcg
    16021 agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata gcatcggtaa
    16081 catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc ggactgatgg
    16141 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
    16201 tgg
//
