LOCUS       sequence-262634-       16388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting A. thaliana pds3, regulated by A. thaliana U6 polIII
            promoter. Cas9 with a C-terminal mApple fusion, egg cell-specific 
            expression. Hygromycin selectable marker (hpt)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16388)
  AUTHORS   Behrendorff JBYH, Borras-Gas G, Pribil M
  TITLE     Antimicrobial solid media for screening non-sterile Arabidopsis 
            thaliana seeds.
  JOURNAL   Physiol Plant. 2020 Feb 25. doi: 10.1111/ppl.13079.
  PUBMED    32096870
REFERENCE   2  (bases 1 to 16388)
  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: "Physiol 
            Plant. 2020 Feb 25. doi: 10.1111/ppl.13079."
COMMENT     Sequence Label: GS2.1/EC
FEATURES             Location/Qualifiers
     source          1..16388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      969..1163
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(1399..1421)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1507..1647
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1562..1581
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1662..1679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1833..2421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1913..1932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2667..3458)
                     /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    3979..4003
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(4096..4348)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(4546..5571)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(5607..5786)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        5958..6261
                     /label=AtU6-1 promoter
                     /note="promoter for the Arabidopsis thaliana U6-1 snRNA 
                     gene (Waibel and Filipowicz, 1990)"
     misc_RNA        6282..6357
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(6369..6393)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6451..6468)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6451..6467)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6460..6482)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    6772..6796
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(6829..7539)
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7107..7126)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(7555..7602)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(join(7603..10266,10456..11892))
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     intron          10267..10455
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             complement(11917..11937)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(11944..11967)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(13461..13485)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    13805..13829
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             15160..15789
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             join(16218..16388,1..903)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="VSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga cgagcaacca
       61 gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag catcatggac
      121 gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat ccgctacgag
      181 cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag tgtgtgggat
      241 tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg ataccgggaa
      301 gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt actcaagttc
      361 tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg cattcggtta
      421 aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg cctggtgacg
      481 gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga aaccgggcgg
      541 ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac agaaggcaag
      601 aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg catcggccgt
      661 tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg gttgttcaag
      721 acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt caccgtgcgc
      781 aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc ggggcaggct
      841 ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc cgccggttcc
      901 taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg tcgaaaaggt
      961 ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg gaaccggaac
     1021 ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta agtgactgat
     1081 ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat taaaactctt
     1141 aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga gctgcaaaaa
     1201 gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg gcctatcgcg
     1261 gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg gcgcggacaa
     1321 gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc tcgcgcgttt
     1381 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     1441 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     1501 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     1561 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1621 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     1681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2401 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2461 tggaacgaaa actcacgtta agggattttg gtcatgcatg atatatctcc caatttgtgt
     2521 agggcttatt atgcacgctt aaaaataata aaagcagact tgacctgata gtttggctgt
     2581 gagcaattat gtgcttagtg catctaacgc ttgagttaag ccgcgccgcg aagcggcgtc
     2641 ggcttgaacg aatttctagc tagacattat ttgccgacta ccttggtgat ctcgcctttc
     2701 acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt
     2761 ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc
     2821 attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa
     2881 atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc
     2941 catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag
     3001 agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag
     3061 atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc
     3121 tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg
     3181 tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa
     3241 gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc
     3301 gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac
     3361 aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat
     3421 agttgagtcg atacttcggc gatcaccgct tcccccatga tgtttaactt tgttttaggg
     3481 cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     3541 taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa
     3601 caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac
     3661 cagttgcgtg agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc
     3721 cactgggttc gtgcccgaat tgatcacagg cagcaacgct ctgtcatcgt tacaatcaac
     3781 atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc
     3841 gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc
     3901 gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg
     3961 gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca
     4021 acttaataac acattgcgga cgtttttaat gtactgaatt aacgccgaat tgaattatca
     4081 gcttgcatgc cggtcgatct agtaacatag atgacaccgc gcgcgataat ttatcctagt
     4141 ttgcgcgcta tattttgttt tctatcgcgt attaaatgta taattgcggg actctaatca
     4201 taaaaaccca tctcataaat aacgtcatgc attacatgtt aattattaca tgcttaacgt
     4261 aattcaacag aaattatatg ataatcatcg caagaccggc aacaggattc aatcttaaga
     4321 aactttattg ccaaatgttt gaacgatctg cttgactcta gggaattaat tcctgaatca
     4381 ctgcgaccgg ccctcccgcg acccagccga gcgagcttag cgaactgtgg acgagaactg
     4441 tgccaccaag cgtaaggccg ttctctcgca tttgccttgc taggctcgcg cgagttgctg
     4501 gctgaggcgt tctcgaaatc agctcttgtt cggtcggcat ctactctatt cctttgccct
     4561 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt
     4621 ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg
     4681 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa
     4741 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc ttggcgatcc
     4801 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca
     4861 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct
     4921 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc
     4981 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag
     5041 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg
     5101 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg
     5161 tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatag catccatagc
     5221 ctccgcgacc ggctgaagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac
     5281 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaactccc caatgtcaag
     5341 cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta
     5401 gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct
     5461 gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt
     5521 ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca taggggggat
     5581 tcgagttgag agtgaatatg agactctaat tggataccga ggggaattta tggaacgtca
     5641 gtggagcatt tttgacaaga aatatttgct agctgatagt gaccttaggc gacttttgaa
     5701 cgcgcaataa tggtttctga cgtatgtgct tagctcatta aactccagaa acccgcggct
     5761 cagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc ttgtcccgcg
     5821 tcatcggcgg gggtcataac gtgactccct taattctccg ctcatgatcg ataattcgag
     5881 ctctcccata tggtcgccat cgctacctta ggaccgttat agttaattac cctgttatcc
     5941 ctattaatta agagctcaga aatctcaaaa ttccggcaga acaattttga atctcgatcc
     6001 gtagaaacga gacggtcatt gttttagttc caccacgatt atatttgaaa tttacgtgag
     6061 tgtgagtgag acttgcataa gaaaataaaa tctttagttg ggaaaaaatt caataatata
     6121 aatgggcttg agaaggaagc gagggatagg cctttttcta aaataggccc atttaagcta
     6181 ttaacaatct tcaaaagtac cacagcgctt aggtaaagaa agcagctgag tttatatatg
     6241 gttagagacg aagtagtgat tggacttttg ccagccatgg tgttttagag ctagaaatag
     6301 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     6361 ttttctagac ccagctttct tgtacaaagt tggcattacc catcggcgcg cctctagaat
     6421 ttaaatggat cctacgtact cgaggaattc actggccgtc gttttacaac gtcgtgactg
     6481 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccaaggt
     6541 cactggattt tggttttagg aattagaaat tttattgata gaagtatttt acaaatacaa
     6601 atacatacta agggtttctt atatgctcaa cacatgagcg aaaccctata agaaccctaa
     6661 ttcccttatc tgggaactac tcacacatta ttctggagaa aaatagagag agatagattt
     6721 gtagagagag actggtgatt tttgcggact ctagcatggc cgcgggatat caccactttg
     6781 tacaagaaag ctgaacgact tctctcatcc gccaaaacag ccaagctttt acttgtacag
     6841 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     6901 gtagtcctcg ttgtgggaca cgatgtccaa cttgatgtcg acgatgtagg cgccgggcag
     6961 ctgcacgggc ttcttggcct tgtaggtggt cttgacctcg gcggcgtagt ggccgccgtc
     7021 cttcagcttc agcctcttct tgatctcgct cttcagggcg ccgtcctcgg ggtacatccg
     7081 ctcctcggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     7141 gttggtgccg cgcagcttca ccttgtagat gaacacgccg tcctgcaggg aggagtcctg
     7201 gttaacgtga ataatgccgc cgtcctcgaa gttcatcacg cgctcccacc tgaagccctc
     7261 ggggaaggac agcttgaagt agtcggggat gtcggctggg tgcttaatgt agaccttgga
     7321 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     7381 caccttcagc ttagcggtct gaaaggcctc gtaggggcgg ccctcgccct cgccctcgat
     7441 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     7501 gatgatggcc atgttattct cctcgccctt gctcaccata gaacctccac ctcccttctt
     7561 cttcttagcc tgtccagcct tcttggtagc agctggtctc ttatctcctc caagctgaga
     7621 aagatcgatt ctggtctcgt aaagtccggt gatagactgg tggataaggg tagcatcaag
     7681 aacctccttg gtagaggtgt atctttttct atcgatggtg gtatcgaagt acttgaaagc
     7741 agctggagca ccaaggttgg taagggtgaa aaggtggatg atgttctcag cctgctctct
     7801 gattggctta tctctgtgct tgttgtaagc agaaagaacc ttatcaaggt tagcatcagc
     7861 aaggataact ctcttagaga actcagagat ttgctcgatg atctcatcaa ggtagtgctt
     7921 gtgctgctca acgaaaagct gcttctgctc gttatcctct ggagatccct tcaacttctc
     7981 gtagtgagaa gcaaggtaaa ggaagttaac gtacttagat ggaagagcaa gctcgtttcc
     8041 cttctgaagc tctccagcag aagcaagcat tctctttctt ccgttctcaa gctcgaaaag
     8101 agagtacttt ggcaacttga tgataagatc cttcttaacc tccttgtatc ccttagcctc
     8161 aaggaaatcg attgggttct tctcgaaaga agaacgctcc atgatggtga ttccaagaag
     8221 ctccttaaca gacttcaact tcttagactt tcccttctca accttagcaa caacaagaac
     8281 agagtaagca acggttggag aatcgaatcc tccgtacttc tttgggtccc aatccttctt
     8341 tctagcgatc aacttatcag agtttctctt tggaaggata gactccttag agaatcctcc
     8401 ggtctgaacc tcggttttct taacgatgtt aacctgtggc atagaaagaa cctttctaac
     8461 ggtagcgaaa tctcttccct tatcccaaac gatctctccg gtctctccgt tggtctcgat
     8521 aagtggtctc tttctgatct ctccgttagc aagggtgatc tcggtcttga agaagttcat
     8581 gatgttagag tagaagaagt acttagcggt agcctttccg atctcctgct cagacttagc
     8641 gatcatcttt ctaacatcgt aaaccttgta atctccgtaa acgaactcag actccaactt
     8701 tgggtacttc ttgataagag cggttccaac aacagcgtta aggtaagcat cgtgagcgtg
     8761 gtggtagttg ttgatctctc taaccttgta gaactggaaa tcctttctga aatcagaaac
     8821 caacttagac ttcaaggtga taaccttaac ctctctgatc aacttatcgt tctcatcgta
     8881 cttggtgttc atacgagaat caaggatctg agcaacgtgc ttggtgatct gtctggtctc
     8941 aacaagctgt ctcttgatga atccagcctt atcaagctca gaaagtcctc ctctctcagc
     9001 cttggtaagg ttatcgaact ttctctgggt gatcaactta gcgttaagaa gctgtctcca
     9061 gtagttcttc atcttcttaa caacctcctc agatggaacg ttatcagact ttcctctgtt
     9121 cttatcagaa cgggtaagaa ccttgttatc gatagaatca tccttcaaga aagactgtgg
     9181 aacgatgtga tcaacatcgt aatcagaaag tctgttgatg tcaagctcct gatcaacgta
     9241 catatctctt ccgttctgaa ggtagtaaag gtacaacttc tcgttctgaa gctgggtgtt
     9301 ctcaactggg tgctccttca agatttgaga tccaagctcc ttgattccct cctcgattct
     9361 cttcattctc tcacgagagt tcttctgtcc cttctgggtg gtctggttct ctctagccat
     9421 ctcgataacg atgttctctg gcttgtgtct acccataacc ttaacaagct catcaacaac
     9481 cttaacggtc tgaaggattc ccttcttgat agctggagat ccagcaaggt tagcgatgtg
     9541 ctcgtgaaga gaatctccct gtccagaaac ctgagccttc tggatgtcct ccttgaaggt
     9601 aagagaatca tcgtggataa gctgcatgaa gtttctgtta gcgaatccat cagacttcaa
     9661 gaaatcaagg atggtctttc cagactgctt atctctgatt ccgttgatca acttacgaga
     9721 aagtctaccc catccggtgt atcttcttct cttcaactgc ttcataacct tatcatcgaa
     9781 aaggtgagcg taggtcttga gtctctcctc gatcatctct ctatcctcga aaagggtaag
     9841 ggtaagaacg atgtcctcaa ggatgtcctc gttctcctcg ttatcaagga aatccttatc
     9901 cttgatgatc ttcaaaagat cgtggtaggt tccaagagaa gcgttgaatc tatcctcaac
     9961 tccagagatt tcaacagaat cgaagcactc gatcttcttg aagtaatcct ccttcaactg
    10021 cttaacggta acctttctgt tggtcttgaa aagaagatca acgatagcct tcttctgctc
    10081 tccagaaagg aaagctggct ttctcattcc ctcggtaacg tacttaacct tggtaagctc
    10141 gttgtaaacg gtgaagtact cgtaaagaag agagtgcttt ggaagaacct tctcgtttgg
    10201 aaggttctta tcgaagttgg tcattctctc gatgaaagac tgagcagaag ctcccttatc
    10261 aacaacctgc acatcaacaa attttggtca tatattagaa aagttataaa ttaaaatata
    10321 cacacttata aactacagaa aagcaattgc tatatactac attcttttat tttgaaaaaa
    10381 atatttgaaa tattatatta ctactaatta atgataatta ttatatatat atcaaaggta
    10441 gaagcagaaa cttacctcct cgaagttcca aggggtgatg gtctcctcag actttctggt
    10501 catccaagcg aaacgagagt ttcctctagc aagtggtcca acgtagtatg ggattctgaa
    10561 ggtaaggatc ttctcgatct tctctctgtt atccttcaag aatgggtaga aatcctcctg
    10621 tctacgaagg atagcgtgaa gctctccaag gtggatctgg tgtgggatag atccgttatc
    10681 gaaggttctc tgctttctaa gaagatcctc tctgttcaac ttaacaagaa gctcctcggt
    10741 tccatccatc ttctcaagga ttggcttgat gaacttgtag aactcctcct gagaagctcc
    10801 tccatcgatg tatccagcgt atccgttctt agactgatcg aagaagattt ccttgtactt
    10861 ctctggaagc tgctgtctaa caagagcctt caaaagggta agatcctggt ggtgctcatc
    10921 gtatctcttg atcatagaag cagaaagtgg agccttggtg atctcggtgt taactctaag
    10981 gatgtcagaa agaaggatag catcagaaag gttcttagca gcaaggaaaa gatcagcgta
    11041 ctgatctccg atctgagcaa gaaggttatc aagatcatca tcgtaggtat ccttagaaag
    11101 ctgcaactta gcatcctcag caagatcgaa gttagacttg aagtttgggg taagtccaag
    11161 agaaagagcg ataaggtttc cgaaaagtcc gttcttcttc tctcctggaa gctgagcgat
    11221 aaggttctca agtctacgag acttagaaag tctagcagaa aggatagcct tagcatcaac
    11281 tccagaagcg ttgattgggt tctcctcgaa aagctggttg taggtctgaa caagctggat
    11341 gaacaactta tcaacatcag agttatctgg gttaaggtct ccctcgataa ggaagtgtcc
    11401 tctgaacttg atcatgtgag caagagcaag gtagataagt ctaagatcag ccttatcggt
    11461 agaatcaacc aacttctttc taaggtggta gatggttggg tacttctcgt ggtaagcaac
    11521 ctcatcaacg atgtttccga agattgggtg tctctcgtgc ttcttatcct cctcaacaag
    11581 gaaagactcc tcaagtctgt ggaagaaaga atcatcaacc ttagccatct cgttagagaa
    11641 gatttcctga aggtagcaga ttctgttctt tcttctggtg tatcttcttc tagcggttct
    11701 cttcaatctg gtagcctcag cggtctctcc agaatcgaaa agaagagcac cgataaggtt
    11761 cttcttgata gagtgtctat cggtgtttcc aagaaccttg aacttcttag atggaacctt
    11821 gtactcatcg gtgataacag cccatccaac agagttggtt ccgatgtcaa gtccgataga
    11881 gtacttctta tcagcagctg gaactccgtg gattccaacc tttctcttct tctttggagc
    11941 catcttatca tcatcatcct tgtaatccat ttctcaacag attgataagg tcgaaagaag
    12001 aaagagtaga agttgctttg gttgatttgt ttgtgatgag aattgtgtgt taactttaga
    12061 gagtttatat agagacagta gagacttgac tgttacaaat aaatggtaga gtgaaagccc
    12121 aagtaattac gagggaagct cattagattg tgagataatt agtgcttagt ggtttagtgg
    12181 gtaaactagg tgtaattact ctattaaaag ctgcgtaatt agtgggtctg tttaggggct
    12241 aatgggccgg attagaatca ctcagtctga aactcgtttg atttagttca atagaaccaa
    12301 atcagcttta ccatcaaaat ctttgcgaga aaatagattt gggtaagtaa agtatgtaca
    12361 catctaatga gcatgtgtga gcatgaaatc tataactatt tgatcatctt gaaaccgaag
    12421 aaatcataag gcaactattt taaagacaca gagacacctg caatactgca aatttgaaag
    12481 gttgctagat ttagtagagc taattcatga taggcgttac tagcttagtg gtgatttaag
    12541 agtaattaaa aagttggatt agtgggagga atgggaaagg attgacggta aatagtcact
    12601 taatcatgaa acgttaaagc atttgactgt agaaacgtat ttagtatata ttggcatgtt
    12661 gaatcagtag cgctcctatt ggagacgaag gtttaacgtc gtgttacaga aagtgaacca
    12721 atgcttttgt aataaactca ttgaacttta tgctatatac tgtatatggt ttttactact
    12781 cattgaactt tatgctatgt tttctttaaa ctccttgtat aaatatattt cttaggaatt
    12841 acttttaaat tatgtacaaa gacatgtatt acagaaaaat ttcatgttgt aatctaaata
    12901 agatgctata gtgttgtcga tgtgtcatgt gtgtcactat ccatcattaa acatgtggta
    12961 aaattacagt ctataaccat catctagatc agatatatct ttttaagaat ttaaacttat
    13021 ggtcaaattt tcacaatctc gaatacaaat atacaagaat gtatattctc aagtctccta
    13081 gtttctctgt tatttgcatc ttttgtgact tacaaactat gataactttt ataaaataga
    13141 tataattagc tatcttcttg gacaagttac ctctcttagg ttgtttctgt tgtttactat
    13201 gctctgattc ttaacaaaaa atggtttatt ttgttctgat aatggtttta agctattcca
    13261 gctcagtgga tctctgtcct tgtataaacg caatgataaa ccaaacgcaa atgcttttat
    13321 tctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
    13381 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
    13441 gcatacgtca aagcaacctc gttcagcttt cttgtacaaa gtggtgatat cccgcggcca
    13501 tgctagagtc cgcaaaaatc accagtctct ctctacaaat ctatctctct ctatttttct
    13561 ccagaataat gtgtgagtag ttcccagata agggaattag ggttcttata gggtttcgct
    13621 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
    13681 aataaaattt ctaattccta aaaccaaaat ccagtgacct gcaggcatgc gacgtcgggc
    13741 ccaagcttag cttgagcttg gatcagattg tcgtttcccg ccttcagttt aaactatcag
    13801 tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc
    13861 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac
    13921 cacagggttc ccctcgggat caaagtactt taaagtactt taaagtactt taaagtactt
    13981 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt
    14041 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
    14101 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
    14161 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
    14221 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
    14281 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
    14341 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
    14401 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
    14461 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
    14521 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
    14581 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
    14641 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
    14701 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
    14761 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
    14821 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
    14881 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
    14941 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
    15001 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
    15061 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
    15121 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
    15181 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
    15241 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
    15301 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
    15361 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
    15421 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
    15481 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
    15541 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
    15601 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
    15661 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
    15721 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
    15781 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
    15841 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
    15901 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
    15961 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
    16021 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
    16081 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
    16141 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
    16201 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
    16261 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
    16321 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
    16381 aatcccgg
//
