LOCUS       sequence-331053-       17142 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Final destination vector for single Gateway LR reaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17142)
  AUTHORS   Ursache R, Fujita S, Denervaud Tendon V, Geldner N
  TITLE     Combined fluorescent seed selection and multiplex CRISPR/Cas9 
            assembly for fast generation of multiple Arabidopsis mutants.
  JOURNAL   Plant Methods. 2021 Oct 30;17(1):111. doi: 
            10.1186/s13007-021-00811-9.
  PUBMED    34717688
REFERENCE   2  (bases 1 to 17142)
  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: "Plant 
            Methods."; date: "2021-10-30"; volume: "17(1):111. doi"; pages: " 
            10.1186/s13007-021-00811-9"
COMMENT     Sequence Label: pRU322
FEATURES             Location/Qualifiers
     source          1..17142
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1187..1816
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2245..3318
                     /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      3384..3578
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(3814..3836)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3922..4062
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3977..3996
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4077..4094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4248..4836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4328..4347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5082..5873)
                     /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    6394..6418
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6595..6616
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6631..6636,6637..6654,6655..6661)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6669..6685
                     /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     6674..6696
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6693..6709
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6693..6709
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             8123..11281
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     CDS             11294..11314
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    11826..11950
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(11975..11980,11981..11998,11999..12005)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             12059..12739
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(12125..12144)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             13059..13364
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     13243..13262
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(13405..13529)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(15783..15785,15786..15788,15789..16502)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(15783..15785,15786..15788,15789..16499)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(15828..15849)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(16089..16108)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     16436..16457
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      complement(16501..16751)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(16762..16779)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(16762..16778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16771..16793)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    16974..16998
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag
       61 ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg
      121 cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac
      181 tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct gggctatgcc
      241 cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca cgcggccggc
      301 tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc
      361 aggatgcttg accacctacg ccctggcgac gttgtgacag tgaccaggct agaccgcctg
      421 gcccgcagca cccgcgacct actggacatt gccgagcgca tccaggaggc cggcgcgggc
      481 ctgcgtagcc tggcagagcc gtgggccgac accaccacgc cggccggccg catggtgttg
      541 accgtgttcg ccggcattgc cgagttcgag cgttccctaa tcatcgaccg cacccggagc
      601 gggcgcgagg ccgccaaggc ccgaggcgtg aagtttggcc cccgccctac cctcaccccg
      661 gcacagatcg cgcacgcccg cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg
      721 gctgcactgc ttggcgtgca tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa
      781 gtgacgccca ccgaggccag gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc
      841 gacgccctgg cggccgccga gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg
      901 acggccagga cgaaccgttt ttcattaccg aagagatcga ggcggagatg atcgcggccg
      961 ggtacgtgtt cgagccgccc gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg
     1021 gtttgtctga tgccaagctg gcggcctggc cggccagctt ggccgctgaa gaaaccgagc
     1081 gccgccgtct aaaaaggtga tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg
     1141 cgtatatgat gcgatgagta aataaacaaa tacgcaaggg gaacgcatga aggttatcgc
     1201 tgtacttaac cagaaaggcg ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc
     1261 cctgcaactc gccggggccg atgttctgtt agtcgattcc gatccccagg gcagtgcccg
     1321 cgattgggcg gccgtgcggg aagatcaacc gctaaccgtt gtcggcatcg accgcccgac
     1381 gattgaccgc gacgtgaagg ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc
     1441 ccaggcggcg gacttggctg tgtccgcgat caaggcagcc gacttcgtgc tgattccggt
     1501 gcagccaagc ccttacgaca tatgggccac cgccgacctg gtggagctgg ttaagcagcg
     1561 cattgaggtc acggatggaa ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg
     1621 cacgcgcatc ggcggtgagg ttgccgaggc gctggccggg tacgagctgc ccattcttga
     1681 gtcccgtatc acgcagcgcg tgagctaccc aggcactgcc gccgccggca caaccgttct
     1741 tgaatcagaa cccgagggcg acgctgcccg cgaggtccag gcgctggccg ctgaaattaa
     1801 atcaaaactc atttgagtta atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta
     1861 agtgccggcc gtccgagcgc acgcagcagc aaggctgcaa cgttggccag cctggcagac
     1921 acgccagcca tgaagcgggt caactttcag ttgccggcgg aggatcacac caagctgaag
     1981 atgtacgcgg tacgccaagg caagaccatt accgagctgc tatctgaata catcgcgcag
     2041 ctaccagagt aaatgagcaa atgaataaat gagtagatga attttagcgg ctaaaggagg
     2101 cggcatggaa aatcaagaac aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg
     2161 aacgggcggt tggccaggcg taagcggctg ggttgtctgc cggccctgca atggcactgg
     2221 aacccccaag cccgaggaat cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc
     2281 ggcgcggcgc tgggtgatga cctggtggag aagttgaagg ccgcgcaggc cgcccagcgg
     2341 caacgcatcg aggcagaagc acgccccggt gaatcgtggc aagcggccgc tgatcgaatc
     2401 cgcaaagaat cccggcaacc gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag
     2461 ggcgacgagc aaccagattt tttcgttccg atgctctatg acgtgggcac ccgcgatagt
     2521 cgcagcatca tggacgtggc cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag
     2581 gtgatccgct acgagcttcc agacgggcac gtagaggttt ccgcagggcc ggccggcatg
     2641 gccagtgtgt gggattacga cctggtactg atggcggttt cccatctaac cgaatccatg
     2701 aaccgatacc gggaagggaa gggagacaag cccggccgcg tgttccgtcc acacgttgcg
     2761 gacgtactca agttctgccg gcgagccgat ggcggaaagc agaaagacga cctggtagaa
     2821 acctgcattc ggttaaacac cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac
     2881 ggccgcctgg tgacggtatc cgagggtgaa gccttgatta gccgctacaa gatcgtaaag
     2941 agcgaaaccg ggcggccgga gtacatcgag atcgagctag ctgattggat gtaccgcgag
     3001 atcacagaag gcaagaaccc ggacgtgctg acggttcacc ccgattactt tttgatcgat
     3061 cccggcatcg gccgttttct ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc
     3121 agatggttgt tcaagacgat ctacgaacgc agtggcagcg ccggagagtt caagaagttc
     3181 tgtttcaccg tgcgcaagct gatcgggtca aatgacctgc cggagtacga tttgaaggag
     3241 gaggcggggc aggctggccc gatcctagtc atgcgctacc gcaacctgat cgagggcgaa
     3301 gcatccgccg gttcctaatg tacggagcag atgctagggc aaattgccct agcaggggaa
     3361 aaaggtcgaa aaggtctctt tcctgtggat agcacgtaca ttgggaaccc aaagccgtac
     3421 attgggaacc ggaacccgta cattgggaac ccaaagccgt acattgggaa ccggtcacac
     3481 atgtaagtga ctgatataaa agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa
     3541 cttattaaaa ctcttaaaac ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc
     3601 gaagagctgc aaaaagcgcc tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg
     3661 cgtcggccta tcgcggccgc tggccgctca aaaatggctg gcctacggcc aggcaatcta
     3721 ccagggcgcg gacaagccgc gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc
     3781 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3841 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3901 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     3961 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     4021 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     4081 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4141 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4201 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4261 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4321 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4381 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4441 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4501 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4561 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4621 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4681 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4741 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4801 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4861 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gcatgatata
     4921 tctcccaatt tgtgtagggc ttattatgca cgcttaaaaa taataaaagc agacttgacc
     4981 tgatagtttg gctgtgagca attatgtgct tagtgcatct aacgcttgag ttaagccgcg
     5041 ccgcgaagcg gcgtcggctt gaacgaattt ctagctagac attatttgcc gactaccttg
     5101 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     5161 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     5221 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     5281 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     5341 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     5401 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     5461 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     5521 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     5581 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     5641 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     5701 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     5761 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     5821 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttcccc catgatgttt
     5881 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     5941 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacccca
     6001 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     6061 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag cttacgaacc
     6121 gaacaggctt atgtccactg ggttcgtgcc cgaattgatc acaggcagca acgctctgtc
     6181 atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac ccggcagctt
     6241 agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc
     6301 tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg attttgtgcc
     6361 gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa
     6421 attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact gaattaacgc
     6481 cgaattgctc tagccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     6541 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     6601 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     6661 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
     6721 ttcggtaccg aataaaagca tttgcgtttg gtttatcatt gcgtttatac aaggacagag
     6781 atccactgag ctggaatagc ttaaaaccat tatcagaaca aaataaacca ttttttgtta
     6841 agaatcagag catagtaaac aacagaaaca acctaagaga ggtaacttgt ccaagaagat
     6901 agctaattat atctatttta taaaagttat catagtttgt aagtcacaaa agatgcaaat
     6961 aacagagaaa ctaggagact tgagaatata cattcttgta tatttgtatt cgagattgtg
     7021 aaaatttgac cataagttta aattcttaaa aagatatatc tgatctaggt gatggttata
     7081 gactgtaatt ttaccacatg tttaatgatg gatagtgaca cacatgacac atcgacaaca
     7141 ctatagcatc ttatttagat tacaacatga aatttttctg taatacatgt ctttgtacat
     7201 aatttaaaag taattcctaa gaaatatatt tatacaagga gtttaaagaa aacatagcat
     7261 aaagttcaat gagtagtaaa aaccatatac agtatatagc ataaagttca atgagtttat
     7321 tacaaaagca ttggttcact ttctgtaaca cgacgttaaa ccttcgtctc caataggagc
     7381 gctactgatt caacatgcca atatatacta aatacgtttc tacagtcaaa tgctttaacg
     7441 tttcatgatt aagtgactat ttaccgtcaa tcctttccca ttcctcccac taatccaact
     7501 ttttaattac tcttaaatca ccactaagct agtaacgcct atcatgaatt agctctacta
     7561 aatctagcaa cctttcaaat ttgcagtatt gcaggtgtct ctgtgtcttt aaaatagttg
     7621 ccttatgatt tcttcggttt caagatgatc aaatagttat agatttcatg ctcacacatg
     7681 ctcattagat gtgtacatac tttacttacc caaatctatt ttctcgcaaa gattttgatg
     7741 gtaaagctga tttggttcta ttgaactaaa tcaaacgagt ttcagactga gtgattctaa
     7801 tccggcccat tagcccctaa acagacccac taattacgca gcttttaata gagtaattac
     7861 acctagttta cccactaaac cactaagcac taattatctc acaatctaat gagcttccct
     7921 cgtaattact tgggctttca ctctaccatt tatttgtaac agtcaagtct ctactgtctc
     7981 tatataaact ctctaaagtt aacacacaat tctcatcaca aacaaatcaa ccaaagcaac
     8041 ttctactctt tcttctttcg accttatcaa tctgttgaga aggtaccaat tcggcgcgcc
     8101 tgtgcagcga attcggcgcg ccatgaagag gaactacatc ctcggactcg atatcggaat
     8161 cacctctgtg ggatacggaa tcatcgatta cgagactagg gatgtgatcg atgctggtgt
     8221 gagactcttc aaagaggcta acgttgagaa caacgaggga agaaggtcta agaggggagc
     8281 tagaaggctc aagagaagaa gaaggcacag aatccagagg gtgaagaagc tcctcttcga
     8341 ttacaacctc ctcaccgatc actctgagct ttctggaatc aacccttacg aggcaagagt
     8401 gaagggactc tctcagaagt tgtctgagga agagttctct gctgctttgc tccaccttgc
     8461 taagagaagg ggtgtgcata acgtgaacga ggtggaagag gataccggaa acgagctttc
     8521 taccaaagag cagatctcta ggaactctaa ggctctcgaa gagaagtacg tggcagagct
     8581 tcagcttgag agactcaaga aagatggtga ggtgagggga tctatcaaca ggttcaagac
     8641 ctctgattac gtgaaagaag ctaagcagct cctcaaggtg cagaaggctt accatcagct
     8701 cgatcagtct ttcatcgata cctacatcga tcttctcgag actagaagga cctactacga
     8761 gggacctggt gaaggatctc cattcggatg gaaggatatc aaagaatggt acgagatgct
     8821 catgggacac tgcacttact tccctgaaga actcagatct gtgaagtacg cttacaacgc
     8881 tgatctctac aacgctctta acgatctcaa caacctcgtg atcaccaggg atgagaacga
     8941 gaagcttgag tactacgaga agttccagat catcgagaac gtgttcaagc aaaagaagaa
     9001 gcctaccctc aagcagatcg ctaaagagat ccttgttaac gaggaagata tcaagggata
     9061 cagggtgacc tctaccggaa agcctgagtt cactaacctc aaggtttacc acgatattaa
     9121 ggatattacc gctagaaaag agattattga gaacgctgag cttctcgatc aaatcgctaa
     9181 gatccttacc atctaccagt catctgagga tatccaagag gaattgacca acctcaactc
     9241 agagcttacc caagaagaga tcgagcagat cagtaacttg aagggttaca ccggaaccca
     9301 caacctctca ctcaaggcta tcaacctcat cctcgatgag ctttggcaca ccaacgataa
     9361 ccagatcgca atcttcaaca gactcaagct cgtgcctaag aaagtggatc tctctcagca
     9421 gaaagagatt cctaccaccc tcgtggatga tttcatcctc tcacctgttg tgaagagatc
     9481 tttcatccag tctatcaagg tgatcaacgc tattatcaag aaatacggac tccctaacga
     9541 tatcatcatc gagttggcta gggaaaagaa ctcaaaggat gctcaaaaga tgatcaacga
     9601 gatgcagaag agaaacaggc agaccaacga gaggatcgaa gagatcatca ggaccaccgg
     9661 aaaagagaac gctaagtacc tcattgagaa gatcaagctc cacgatatgc aagagggaaa
     9721 gtgcctctac tctctcgagg ctatccctct tgaggatctc cttaacaacc ctttcaacta
     9781 cgaagtggat cacatcatcc ctagatctgt gtctttcgat aactctttca acaacaaggt
     9841 gctcgtgaag caagaggaaa actctaagaa gggtaacagg acccctttcc agtacctctc
     9901 ttcatctgat tctaagatct catacgagac tttcaagaag cacatcctca acctcgctaa
     9961 gggaaagggt aggatctcta agacaaagaa agagtacctt ctcgaggaaa gggatatcaa
    10021 cagattctct gttcagaagg atttcatcaa cagaaacctt gtggatacca ggtacgctac
    10081 caggggtctt atgaaccttc tcaggtctta cttcagggtg aacaaccttg atgtgaaggt
    10141 gaagtcaatc aacggtggat tcacctcatt ccttagaaga aaatggaagt ttaagaaaga
    10201 gaggaacaag ggttataagc accacgctga ggatgctctc attatcgcta acgcagattt
    10261 cattttcaaa gagtggaaga agttggataa ggctaaaaag gtgatggaaa accagatgtt
    10321 cgaggaaaag caggctgagt ctatgcctga gatcgagact gagcaagagt acaaagaaat
    10381 cttcatcacc cctcaccaga tcaagcacat caaagatttc aaggattaca agtactctca
    10441 cagagtggat aagaagccaa acagagagct tatcaacgat accctctact caaccagaaa
    10501 ggatgataag ggaaacaccc tcatcgtgaa caatctcaac ggactctacg ataaggataa
    10561 cgataagctc aagaagctca ttaacaagtc tccagagaag ttgctcatgt accaccacga
    10621 tcctcagacc taccaaaagc tcaagctcat catggaacag tacggtgatg agaagaaccc
    10681 tctctacaag tattacgagg aaaccggtaa ctacctcacc aagtactcaa aaaaggataa
    10741 tggacctgtg atcaagaaga ttaagtacta cggaaacaag ctcaacgcac acctcgatat
    10801 caccgatgat taccctaaca gtagaaacaa ggtggtgaag ctttcactca agccttacag
    10861 gttcgatgtg tacctcgata acggtgtgta caagttcgtg accgtgaaga acctcgatgt
    10921 tatcaaaaaa gaaaactact atgaggttaa ctctaagtgc tacgaggaag ctaagaagtt
    10981 gaagaagatc tctaaccagg cagagttcat tgcttcattc tacaacaacg atcttatcaa
    11041 gattaacggt gagctttaca gggttatcgg agtgaacaac gatttgctca acaggatcga
    11101 ggtgaacatg attgatatca cctacaggga atacctcgag aatatgaacg ataagaggcc
    11161 tcctagaatc atcaagacta tcgcttctaa gacccagagt atcaagaagt actctaccga
    11221 tatcctcgga aacctctacg aggttaagtc aaagaagcac cctcagatca ttaagaaggg
    11281 atcaagggct gatcctaaga agaagaggaa ggtttgaggc gcgccgagct ccaggcctcc
    11341 cagctttcgt ccgtatcatc ggtttcgaca acgttcgtca agttcaatgc atcagtttca
    11401 ttgcccacac accagaatcc tactaagttt gagtattatg gcattggaaa agctgttttc
    11461 ttctatcatt tgttctgctt gtaatttact gtgttctttc agtttttgtt ttcggacatc
    11521 aaaatgcaaa tggatggata agagttaata aatgatatgg tccttttgtt cattctcaaa
    11581 ttattattat ctgttgtttt tactttaatg ggttgaattt aagtaagaaa ggaactaaca
    11641 gtgtgatatt aaggtgcaat gttagacata taaaacagtc tttcacctct ctttggttat
    11701 gtcttgaatt ggtttgtttc ttcacttatc tgtgtaatca agtttactat gagtctatga
    11761 tcaagtaatt atgcaatcaa gttaagtaca gtataggctt gagctcccta ggcctgttat
    11821 ccctaacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata
    11881 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
    11941 agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
    12001 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
    12061 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
    12121 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
    12181 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
    12241 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
    12301 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
    12361 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
    12421 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
    12481 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
    12541 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
    12601 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
    12661 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcaggcgg ggcgtaatct
    12721 agaggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
    12781 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga
    12841 agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga
    12901 tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc
    12961 cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat
    13021 gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct
    13081 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
    13141 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtccccc
    13201 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
    13261 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
    13321 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta
    13381 tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat
    13441 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
    13501 tctcgttcag ctttcttgta caaagtggtt cgataattcc gatccagcct aggcccgggc
    13561 ctgaggacgc gtccatggtt aattaagacg tccggaccga ctagtggatc ctctagagtc
    13621 gacctgcagg catgcaagct tcttcaagtg tatgtaggta tagtaacatg aacaagaaca
    13681 gactcaagta caagatcgca tacgaaaatg gaaatggcaa tgtcacttcc acataatcaa
    13741 acacgaatcc tcatatcaac aaggcctgag attctaacta gctcataaca acttagccaa
    13801 tagttacttg agactaccaa atgtatgtag aactaaagac taagggacag agagttcgtc
    13861 taaacaggtg aatctagtcg ttgttatcta ataaacaatt cagccccaaa tgcagaacac
    13921 acatagagct ctctattgat tcaaattacg atctgatact gataacgtct agatttttag
    13981 ggttaaagca atcaatcacc tgacgattca aggtggttgg atcatgacga ttccagaaaa
    14041 catcaagcaa gctctcaaag ctacactctt tgggatcata ctgaactcta acaacctcgt
    14101 tatgtcccgt agtgccagta cagacatcct cgtaactcgg attgtgcacg atgccatggc
    14161 tatacccaac ctcggtcttg gtcacaccag gaactctctg gtaagctagc tccactcccc
    14221 agaaacaacc ggcgccaaat tgcgcgaatt gctgacctga agacggaaca tcatcgtcgg
    14281 gtccttgggc gattgcggcg gaagatgggt cagcttgggc ttgaggacga gacccgaatc
    14341 cgagtctgtt gaaaaggttg ttcattgggg atttgtatac ggagattggt cgtcgagagg
    14401 tttgagggaa aggacaaatg ggtttggctc tggagaaaga gagtgcggct ttagagagag
    14461 aattgagagg tttagagaga gatgcggcgg cgatgagcgg aggagagacg acgaggacct
    14521 gcattatcaa agcagtgacg tggtgaaatt tggaactttt aagaggcaga tagatttatt
    14581 atttgtatcc attttcttca ttgttctaga atgtcgcgga acaaatttta aaactaaatc
    14641 ctaaattttt ctaattttgt tgccaatagt ggatatgtgg gccgtataga aggaatctat
    14701 tgaaggccca aacccatact gacgagccca aaggttcgtt ttgcgtttta tgtttcggtt
    14761 cgatgccaac gccacattct gagctaggca aaaaacaaac gtgtctttga atagactcct
    14821 ctcgttaaca catgcagcgg ctgcatggtg acgccattaa cacgtggcct acaattgcat
    14881 gatgtctcca ttgacacgtg acttctcgtc tcctttctta atatatctaa caaacactcc
    14941 tacctcttcc aaaatatata cacatctttt tgatcaatct ctcattcaaa atctcattct
    15001 ctctagtaaa caagaacaaa aaaatggcgg atacagctag aggaacccat cacgatatca
    15061 tcggcagaga ccagtacccg atgatgggcc gagaccgaga ccagtaccag atgtccggac
    15121 gaggatctga ctactccaag tctaggcaga ttgctaaagc tgcaactgct gtcacagctg
    15181 gtggttccct ccttgttctc tccagcctta cccttgttgg aactgtcata gctttgactg
    15241 ttgcaacacc tctgctcgtt atcttcagcc caatccttgt cccggctctc atcacagttg
    15301 cactcctcat caccggtttt ctttcctctg gagggtttgg cattgccgct ataaccgttt
    15361 tctcttggat ttacaagtaa gcacacattt atcatcttac ttcataattt tgtgcaatat
    15421 gtgcatgcat gtgttgagcc agtagctttg gatcaatttt tttggtcgaa taacaaatgt
    15481 aacaataaga aattgcaaat tctagggaac atttggttaa ctaaatacga aatttgacct
    15541 agctagcttg aatgtgtctg tgtatatcat ctatataggt aaaatgcttg gtatgatacc
    15601 tattgattgt gaataggtac gcaacgggag agcacccaca gggatcagac aagttggaca
    15661 gtgcaaggat gaagttggga agcaaagctc aggatctgaa agacagagct cagtactacg
    15721 gacagcaaca tactggtggg gaacatgacc gtgaccgtac tcgtggtggc cagcacacta
    15781 ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
    15841 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
    15901 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
    15961 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
    16021 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
    16081 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
    16141 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
    16201 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
    16261 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
    16321 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
    16381 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
    16441 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
    16501 gatctagtaa catagatgaa ccgcgcgcga taatttatcc tagtttgcgc gctatatttt
    16561 gttttctatc gcgtattaaa tgtataattg cgggactcta atcataaaaa cccatctcat
    16621 aaataacgtc atgcattaca tgttaattat tacatgctta acgtaattca acagaaatta
    16681 tatgataatc atcgcaagac cggcaacagg attcaatctt aagaaacttt attgccaaat
    16741 gtttgaacga tgaagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
    16801 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
    16861 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgagc
    16921 ttgagcttgg atcagattgt cgtttcccgc cttcagttta aactatcagt gtttgacagg
    16981 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
    17041 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
    17101 cctcgggatc aaagtacttt aaagtacttt aaagtacttt aa
//
