LOCUS       sequence-490463-       15191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CaMV 35S driving the expression of dCas9-ZAT10(1x).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15191)
  AUTHORS   Khan MA, Herring G, Zhu JY, Oliva M, Fourie E, Johnston B, Zhang Z, 
            Potter J, Pineda L, Pflueger J, Swain T, Pflueger C, Lloyd JPB, 
            Secco D, Small I, Kidd BN, Lister R
  TITLE     CRISPRi-based circuits to control gene expression in plants.
  JOURNAL   Nat Biotechnol. 2025 Mar;43(3):416-430. doi: 
            10.1038/s41587-024-02236-w. Epub 2024 May 20.
  PUBMED    38769424
REFERENCE   2  (bases 1 to 15191)
  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"; doi: 
            "10.1038/s41587-024-02236-w"; journalName: "Nat Biotechnol"; date: 
            "2025-03"; volume: "43"; issue: "3"; pages: "416-430"
COMMENT     Sequence Label: LLP1019
FEATURES             Location/Qualifiers
     source          1..15191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    62..86
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1417..2046
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2475..3548
                     /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      3614..3808
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4044..4066)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4152..4292
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4207..4226
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4307..4324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4478..5066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4558..4577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5312..6103)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_feature    6624..6648
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(6741..6993)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(7013..7564)
                     /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(7606..7785)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        7907..8251
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    8252..8271
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        8292..8367
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    8368..8387
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     terminator      8417..8617
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     CDS             8983..9003
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9809..9857
                     /note="synthetic polyadenylation signal"
     misc_feature    9871..9962
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     9911..9930
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        9969..10313
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             10329..10349
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(10365..11801,11991..14654)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system"
                     /label=pcoCas9
                     /note="plant codon-optimized Cas9 gene containing the 
                     potato IV2 intron"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     intron          11802..11990
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             14667..14687
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14691..14711
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      14928..15180
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 agcttagctt gagcttggat cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt
       61 ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
      121 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac
      181 agggttcccc tcgggatcaa agtactttaa agtactttaa agtactttaa agtactttga
      241 tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg
      301 aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg cccttttcct
      361 ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac tagaaccgga
      421 gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc cgcgtcagca
      481 ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc tgcaccaagc
      541 tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg
      601 accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg gcccgcagca
      661 cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc
      721 tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg accgtgttcg
      781 ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc gggcgcgagg
      841 ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg gcacagatcg
      901 cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc
      961 ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca
     1021 ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc gacgccctgg
     1081 cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg acggccagga
     1141 cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg ggtacgtgtt
     1201 cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg gtttgtctga
     1261 tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc gccgccgtct
     1321 aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat
     1381 gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac
     1441 cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc
     1501 gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg
     1561 gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc
     1621 gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg
     1681 gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc
     1741 ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc
     1801 acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc
     1861 ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc
     1921 acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa
     1981 cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc
     2041 atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc
     2101 gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca
     2161 tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg
     2221 tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt
     2281 aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa
     2341 aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt
     2401 tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg aacccccaag
     2461 cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc
     2521 tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg
     2581 aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat
     2641 cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc
     2701 aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca
     2761 tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct
     2821 acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt
     2881 gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc
     2941 gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca
     3001 agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc
     3061 ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg
     3121 tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg
     3181 ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag
     3241 gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg
     3301 gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt
     3361 tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg
     3421 tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc
     3481 aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg
     3541 gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa
     3601 aaggtctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc
     3661 ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga
     3721 ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa
     3781 ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc
     3841 aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta
     3901 tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg
     3961 gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg
     4021 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     4081 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     4141 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
     4201 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     4261 acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact
     4321 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4381 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4441 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4501 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4561 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4621 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4681 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4741 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4801 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4861 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4921 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4981 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5041 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5101 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcatgatata tctcccaatt
     5161 tgtgtagggc ttattatgca cgcttaaaaa taataaaagc agacttgacc tgatagtttg
     5221 gctgtgagca attatgtgct tagtgcatct aacgcttgag ttaagccgcg ccgcgaagcg
     5281 gcgtcggctt gaacgaattt ctagctagac attatttgcc gactaccttg gtgatctcgc
     5341 ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt
     5401 cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc
     5461 gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt
     5521 accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg
     5581 agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat
     5641 caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca
     5701 gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat
     5761 tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt
     5821 cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag
     5881 ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg
     5941 tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc
     6001 cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct
     6061 ctgatagttg agtcgatact tcggcgatca ccgcttcccc catgatgttt aactttgttt
     6121 tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca
     6181 cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt
     6241 cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc
     6301 tggaccagtt gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt
     6361 atgtccactg ggttcgtgcc cgaattgatc acaggcagca acgctctgtc atcgttacaa
     6421 tcaacatgct accctccgcg agatcatccg tgtttcaaac ccggcagctt agttgccgtt
     6481 cttccgaata gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc tctcccgctg
     6541 acgccgtccc ggactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg
     6601 tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt
     6661 agacaactta ataacacatt gcggacgttt ttaatgtact gaattaacgc cgaattgaat
     6721 tatcagcttg catgccggtc gatctagtaa catagatgac accgcgcgcg ataatttatc
     6781 ctagtttgcg cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct
     6841 aatcataaaa acccatctca taaataacgt catgcattac atgttaatta ttacatgctt
     6901 aacgtaattc aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct
     6961 taagaaactt tattgccaaa tgtttgaacg atctgcttga ctctaggggt catcagattt
     7021 cggtgacggg caggaccgga cggggcggca ccggcaggct gaagtccagc tgccagaaac
     7081 ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat
     7141 atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca
     7201 cgctcttgaa gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca
     7261 gtcccgtccg ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg
     7321 cgttgcgtgc cttccaggga cccgcgtagg cgatgccggc gacctcgccg tccacctcgg
     7381 cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt
     7441 cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc
     7501 agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc
     7561 tcatggtaga tcccctcgat cgagttgaga gtgaatatga gactctaatt ggataccgag
     7621 gggaatttat ggaacgtcag tggagcattt ttgacaagaa atatttgcta gctgatagtg
     7681 accttaggcg acttttgaac gcgcaataat ggtttctgac gtatgtgctt agctcattaa
     7741 actccagaaa cccgcggctc agtggctcct tcaacgttgc ggttctgtca gttccaaacg
     7801 taaaacggct tgtcccgcgt catcggcggg ggtcataacg tgactccctt aattctccgc
     7861 tcatgatcga taattcgagg gtacaagctt atctaacgat gagctctgag acttttcaac
     7921 aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca
     7981 aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg
     8041 ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
     8101 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
     8161 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta
     8221 tataaggaag ttcatttcat ttggagagga cgttcactgc cgtataggca gtatagaacc
     8281 gatcctccca tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     8341 cttgaaaaag tggcaccgag tcggtgcgtt cactgccgta taggcagctg cagactgact
     8401 aactaggatg agctaagcta gctatatcat caatttatgt attacacata atatcgcact
     8461 cagtctttca tctacggcaa tgtaccagct gatataatca gttattgaaa tatttctgaa
     8521 tttaaacttg catcaataaa tttatgtttt tgcttggact ataatacctg acttgttatt
     8581 ttatcaataa atatttaaac tatatttctt tcaagatggg aattaacatc tacaaattgc
     8641 cttttcttat cgaccatgta ccatatgcca acactcgaat ccccacccgt tggaccaaac
     8701 ccggctcatt aagcgtcggt tcagatttat ttcctttatt taaaaaaaag gaaagggtaa
     8761 aaaatagaaa attggaaaca gttaaagccc aaaattgtaa tttaccgaga attgtaaatt
     8821 tacctgaaaa ccctacgcta tagtttcgac tataaatacc aaacttagga cctcacttca
     8881 gaatcccctc gtcgctgcgt ctctctcccg caaccttcga ttttcgttta ttcgcatcca
     8941 tcggagagag aaaacaatca ataagcgacc cctaggatgg ctccaaaaaa gaaaagaaag
     9001 gttgctagcg accattactt agatattaga ttgagaccag acccagagtt ccctccagca
     9061 cagttgatgt cagtgttatt cggaaaattg caccaggctt tggttgcaca aggtggagat
     9121 agaattggtg tgagttttcc tgatttggac gagtctaggt caagattggg agaaaggtta
     9181 agaatccatg cttcagcaga tgaccttagg gctttgttag caagaccttg gttagagggt
     9241 cttagagatc atttgcagtt cggagaacca gctgttgtgc cacaccctac accatataga
     9301 caagtttcta gagtgcaggc aaaatcaaat cctgagagac ttagaaggag attgatgagg
     9361 agacacgact tatctgaaga ggaagctagg aagagaatac ctgataccgt tgctagagca
     9421 cttgacttgc catttgtgac tttgagaagt caatccactg gtcagcattt taggttattc
     9481 attagacacg gaccacttca agttactgct gaggagggag gattcacctg ttatggattg
     9541 agtaagggag gattcgtgcc ttggttctga ggtacccggc catgctagag tccgcaaaaa
     9601 tcaccagtct ctctctacaa atctatctct ctctattttt ctccagaata atgtgtgagt
     9661 agttcccaga taagggaatt agggttctta tagggtttcg ctcatgtgtt gagcatataa
     9721 gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc
     9781 taaaaccaaa atccagtgac ctactagaaa taaaatatct ttattttcat tacatctgtg
     9841 tgttggtttt ttgtgtgaat cgatagtact aacatacgct ctccatcaaa acaaaacgaa
     9901 acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt gcaggtgcca gaacatttct
     9961 ctgaattctg agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc
    10021 ccagctatct gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc
    10081 catcattgcg ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa
    10141 gatggacccc cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca
    10201 aagcaagtgg attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat
    10261 ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag gacggcacta
    10321 gtgatatgcc aaagaagaag agaaaggtcg gaggaggagg atctgacaag aagtactcta
    10381 tcggacttgc catcggaacc aactctgttg gatgggctgt tatcaccgat gagtacaagg
    10441 ttccatctaa gaagttcaag gttcttggaa acaccgatag acactctatc aagaagaacc
    10501 ttatcggtgc tcttcttttc gattctggag agaccgctga ggctaccaga ttgaagagaa
    10561 ccgctagaag aagatacacc agaagaaaga acagaatctg ctaccttcag gaaatcttct
    10621 ctaacgagat ggctaaggtt gatgattctt tcttccacag acttgaggag tctttccttg
    10681 ttgaggagga taagaagcac gagagacacc caatcttcgg aaacatcgtt gatgaggttg
    10741 cttaccacga gaagtaccca accatctacc accttagaaa gaagttggtt gattctaccg
    10801 ataaggctga tcttagactt atctaccttg ctcttgctca catgatcaag ttcagaggac
    10861 acttccttat cgagggagac cttaacccag ataactctga tgttgataag ttgttcatcc
    10921 agcttgttca gacctacaac cagcttttcg aggagaaccc aatcaacgct tctggagttg
    10981 atgctaaggc tatcctttct gctagacttt ctaagtctcg tagacttgag aaccttatcg
    11041 ctcagcttcc aggagagaag aagaacggac ttttcggaaa ccttatcgct ctttctcttg
    11101 gacttacccc aaacttcaag tctaacttcg atcttgctga ggatgctaag ttgcagcttt
    11161 ctaaggatac ctacgatgat gatcttgata accttcttgc tcagatcgga gatcagtacg
    11221 ctgatctttt ccttgctgct aagaaccttt ctgatgctat ccttctttct gacatcctta
    11281 gagttaacac cgagatcacc aaggctccac tttctgcttc tatgatcaag agatacgatg
    11341 agcaccacca ggatcttacc cttttgaagg ctcttgttag acagcagctt ccagagaagt
    11401 acaaggaaat cttcttcgat cagtctaaga acggatacgc tggatacatc gatggaggag
    11461 cttctcagga ggagttctac aagttcatca agccaatcct tgagaagatg gatggaaccg
    11521 aggagcttct tgttaagttg aacagagagg atcttcttag aaagcagaga accttcgata
    11581 acggatctat cccacaccag atccaccttg gagagcttca cgctatcctt cgtagacagg
    11641 aggatttcta cccattcttg aaggataaca gagagaagat cgagaagatc cttaccttca
    11701 gaatcccata ctacgttgga ccacttgcta gaggaaactc tcgtttcgct tggatgacca
    11761 gaaagtctga ggagaccatc accccttgga acttcgagga ggtaagtttc tgcttctacc
    11821 tttgatatat atataataat tatcattaat tagtagtaat ataatatttc aaatattttt
    11881 ttcaaaataa aagaatgtag tatatagcaa ttgcttttct gtagtttata agtgtgtata
    11941 ttttaattta taacttttct aatatatgac caaaatttgt tgatgtgcag gttgttgata
    12001 agggagcttc tgctcagtct ttcatcgaga gaatgaccaa cttcgataag aaccttccaa
    12061 acgagaaggt tcttccaaag cactctcttc tttacgagta cttcaccgtt tacaacgagc
    12121 ttaccaaggt taagtacgtt accgagggaa tgagaaagcc agctttcctt tctggagagc
    12181 agaagaaggc tatcgttgat cttcttttca agaccaacag aaaggttacc gttaagcagt
    12241 tgaaggagga ttacttcaag aagatcgagt gcttcgattc tgttgaaatc tctggagttg
    12301 aggatagatt caacgcttct cttggaacct accacgatct tttgaagatc atcaaggata
    12361 aggatttcct tgataacgag gagaacgagg acatccttga ggacatcgtt cttaccctta
    12421 cccttttcga ggatagagag atgatcgagg agagactcaa gacctacgct caccttttcg
    12481 atgataaggt tatgaagcag ttgaagagaa gaagatacac cggatggggt agactttctc
    12541 gtaagttgat caacggaatc agagataagc agtctggaaa gaccatcctt gatttcttga
    12601 agtctgatgg attcgctaac agaaacttca tgcagcttat ccacgatgat tctcttacct
    12661 tcaaggagga catccagaag gctcaggttt ctggacaggg agattctctt cacgagcaca
    12721 tcgctaacct tgctggatct ccagctatca agaagggaat ccttcagacc gttaaggttg
    12781 ttgatgagct tgttaaggtt atgggtagac acaagccaga gaacatcgtt atcgagatgg
    12841 ctagagagaa ccagaccacc cagaagggac agaagaactc tcgtgagaga atgaagagaa
    12901 tcgaggaggg aatcaaggag cttggatctc aaatcttgaa ggagcaccca gttgagaaca
    12961 cccagcttca gaacgagaag ttgtaccttt actaccttca gaacggaaga gatatgtacg
    13021 ttgatcagga gcttgacatc aacagacttt ctgattacga tgttgatgct atcgttccac
    13081 agtctttctt gaaggatgat tctatcgata acaaggttct tacccgttct gataagaaca
    13141 gaggaaagtc tgataacgtt ccatctgagg aggttgttaa gaagatgaag aactactgga
    13201 gacagcttct taacgctaag ttgatcaccc agagaaagtt cgataacctt accaaggctg
    13261 agagaggagg actttctgag cttgataagg ctggattcat caagagacag cttgttgaga
    13321 ccagacagat caccaagcac gttgctcaga tccttgattc tcgtatgaac accaagtacg
    13381 atgagaacga taagttgatc agagaggtta aggttatcac cttgaagtct aagttggttt
    13441 ctgatttcag aaaggatttc cagttctaca aggttagaga gatcaacaac taccaccacg
    13501 ctcacgatgc ttaccttaac gctgttgttg gaaccgctct tatcaagaag tacccaaagt
    13561 tggagtctga gttcgtttac ggagattaca aggtttacga tgttagaaag atgatcgcta
    13621 agtctgagca ggagatcgga aaggctaccg ctaagtactt cttctactct aacatcatga
    13681 acttcttcaa gaccgagatc acccttgcta acggagagat cagaaagaga ccacttatcg
    13741 agaccaacgg agagaccgga gagatcgttt gggataaggg aagagatttc gctaccgtta
    13801 gaaaggttct ttctatgcca caggttaaca tcgttaagaa aaccgaggtt cagaccggag
    13861 gattctctaa ggagtctatc cttccaaaga gaaactctga taagttgatc gctagaaaga
    13921 aggattggga cccaaagaag tacggaggat tcgattctcc aaccgttgct tactctgttc
    13981 ttgttgttgc taaggttgag aagggaaagt ctaagaagtt gaagtctgtt aaggagcttc
    14041 ttggaatcac catcatggag cgttcttctt tcgagaagaa cccaatcgat ttccttgagg
    14101 ctaagggata caaggaggtt aagaaggatc ttatcatcaa gttgccaaag tactctcttt
    14161 tcgagcttga gaacggaaga aagagaatgc ttgcttctgc tggagagctt cagaagggaa
    14221 acgagcttgc tcttccatct aagtacgtta acttccttta ccttgcttct cactacgaga
    14281 agttgaaggg atctccagag gataacgagc agaagcagct tttcgttgag cagcacaagc
    14341 actaccttga tgagatcatc gagcaaatct ctgagttctc taagagagtt atccttgctg
    14401 atgctaacct tgataaggtt ctttctgctt acaacaagca cagagataag ccaatcagag
    14461 agcaggctga gaacatcatc caccttttca cccttaccaa ccttggtgct ccagctgctt
    14521 tcaagtactt cgataccacc atcgatagaa aaagatacac ctctaccaag gaggttcttg
    14581 atgctaccct tatccaccag tctatcaccg gactttacga gaccagaatc gatctttctc
    14641 agcttggagg agatgaagga gctgatccta agaagaaaag aaaggttgat ccaaagaaaa
    14701 agaggaaagt tggtggaggt ggatcaggag gtggaggatc tggtggggga ggatccgagg
    14761 gagcaggatc aacttcacat gtgtcttcat ctcacagagg ttttgacctt aacattccac
    14821 ctatccctga gttttcaatg gtgaatggtg atgacgaagt gatgtcacct atgccagcta
    14881 aaaagccaag attcgatttc cctgtgaagt tgcaattgtg agtcgacgat cgttcaaaca
    14941 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgacg attatcatat
    15001 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
    15061 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
    15121 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
    15181 agtcactcga g
//
