LOCUS       sequence-176428-       14841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Disruption of GLABROUS1 gene in Arabidopsis using CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14841)
  AUTHORS   Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP
  TITLE     An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis 
            thaliana.
  JOURNAL   Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. 
            eCollection 2017.
  PUBMED    28174584
REFERENCE   2  (bases 1 to 14841)
  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: "Front Plant
            Sci."; date: "2017-01-24"; volume: "8:39. doi"; pages: " 
            10.3389/fpls.2017.00039. eCollection 2017"
COMMENT     Sequence Label: pUB-Cas9-@GL1
FEATURES             Location/Qualifiers
     source          1..14841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    41..215
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    293..317
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             742..1536
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1623..2211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2112..2131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2365..2382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2397..2537
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2463..2482)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2623..2645
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      2881..3075
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(3141..4214)
                     /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"
     CDS             complement(4643..5272)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    6573..6597
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     6785..6807
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6799..6816
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6800..6816
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      6849..6858
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      6853..6862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_RNA        7359..7434
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    8238..8262
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8283..8303
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8304..12407
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8304..12404
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     protein_bind    complement(12411..12435)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      12499..12751
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12768..12784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12768..12784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12781..12803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12792..12808
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12816..12822,12823..12840,12841..12846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12861..12882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        13073..13749
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     13686..13706
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             13816..14841
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
ORIGIN
        1 agtagatgcc gaccgggatc tgtcgatcga caagctcgag tttctccata ataatgtgtg
       61 agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata
      121 taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat
      181 tcctaaaacc aaaatccagt actaaaatcc agatcccccg aattaattcg gcgttaattc
      241 agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac
      301 cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat
      361 caccactcga tacaggcagc ccatcagtcc gggacggcgt cagcgggaga gccgttgtaa
      421 ggcggcagac tttgctcatg ttaccgatgc tattcggaag aacggcaact aagctgccgg
      481 gtttgaaaca cggatgatct cgcggagggt agcatgttga ttgtaacgat gacagagcgt
      541 tgctgcctgt gatcaccgcg gtttcaaaat cggctccgtc gatactatgt tatacgccaa
      601 ctttgaaaac aactttgaaa aagctgtttt ctggtattta aggttttaga atgcaaggaa
      661 cagtgaattg gagttcgtct tgttataatt agcttcttgg ggtatcttta aatactgtag
      721 aaaagaggaa ggaaataata aatggctaaa atgagaatat caccggaatt gaaaaaactg
      781 atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa ggtatataag
      841 ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta taaagggacc
      901 acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa gctgcctgtt
      961 ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat gagtgaggcc
     1021 gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa gattatcgag
     1081 ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg tccctatacg
     1141 aatagcttag acagccgctt agccgaattg gattacttac tgaataacga tctggccgat
     1201 gtggattgcg aaaactggga agaagacact ccatttaaag atccgcgcga gctgtatgat
     1261 tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga cctgggagac
     1321 agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg gagaagcggc
     1381 agggcggaca agtggtatga cattgccttc tgcgtccggt cgatcaggga ggatatcggg
     1441 gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga ttgggagaaa
     1501 ataaaatatt atattttact ggatgaattg ttttagtacc tagaatgcat gaccaaaatc
     1561 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1621 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1681 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1741 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     1801 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1861 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1921 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1981 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2041 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2101 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2161 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2221 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     2281 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     2341 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     2401 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     2461 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     2521 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     2581 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     2641 agaggttttc accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg
     2701 cggtcgagtg gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccgtaggcca
     2761 gccatttttg agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag
     2821 cgcagcgacc gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt
     2881 tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa
     2941 aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac
     3001 ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg
     3061 tacgtgctat ccacaggaaa gagacctttt cgaccttttt cccctgctag ggcaatttgc
     3121 cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg
     3181 tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc
     3241 aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg
     3301 ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg
     3361 cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg
     3421 tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct
     3481 agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta
     3541 atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc
     3601 tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc
     3661 tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg
     3721 cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa
     3781 accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa
     3841 acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc
     3901 ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca
     3961 tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac
     4021 ggcgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc
     4081 cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc
     4141 ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt
     4201 ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc
     4261 aggcaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca
     4321 cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc
     4381 atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc
     4441 agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc
     4501 gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt
     4561 gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc
     4621 attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct
     4681 ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg
     4741 cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc
     4801 cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga
     4861 caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca
     4921 ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg
     4981 ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga
     5041 agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa
     5101 cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg
     5161 aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga
     5221 tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc
     5281 cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct
     5341 gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa
     5401 gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg
     5461 cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc
     5521 gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt
     5581 tcctcttggc gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca
     5641 cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg
     5701 cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct
     5761 tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg
     5821 ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt
     5881 agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc
     5941 gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg
     6001 cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact
     6061 gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc
     6121 ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc
     6181 cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg
     6241 gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg
     6301 cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta
     6361 acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg tcagaagccg
     6421 actgcactat agcagcggag gggttggatc aaagtacttt gatcccgagg ggaaccctgt
     6481 ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc
     6541 cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac
     6601 actgatagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa gctgctctag
     6661 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     6721 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6781 ttttcccagt cacgacgttg taaaacgacg gccagtgcca gaattgggcc cgacgtcgca
     6841 tgctcccggc cgccatggcg gccgcgggaa ttcgattcac gaagcttcgt tgaacaacgg
     6901 aaactcgact tgccttccgc acaatacatc atttcttctt agcttttttt cttcttcttc
     6961 gttcatacag tttttttttg tttatcagct tacattttct tgaaccgtag ctttcgtttt
     7021 cttcttttta actttccatt cggagttttt gtatcttgtt tcatagtttg tcccaggatt
     7081 agaatgatta ggcatcgaac cttcaagaat ttgattgaat aaaacatctt cattcttaag
     7141 atatgaagat aatcttcaaa aggcccctgg gaatctgaaa gaagagaagc aggcccattt
     7201 atatgggaaa gaacaatagt atttcttata taggcccatt taagttgaaa acaatcttca
     7261 aaagtcccac atcgcttaga taagaaaacg aagctgagtt tatatacagc tagagtcgaa
     7321 gtagtgattg tcccttcggg aaaagttgta gactgagagt tttagagcta gaaatagcaa
     7381 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcaaattt
     7441 ttttaaacta gtggtgatct tgctgaaaaa ctcgagccat ccggaagatc ttgaattcgc
     7501 ggccgcctgc aggtcgacca tatgggagag ctcccaacgc gttggatgca tagcttggta
     7561 cccgacgagt cagtaataaa cggcgtcaaa gtggttgcag ccggcacaca cgagtcgtgt
     7621 ttatcaactc aaagcacaaa tacttttcct caacctaaaa ataaggcaat tagccaaaaa
     7681 caactttgcg tgtaaacaac gctcaataca cgtgtcattt tattattagc tattgcttca
     7741 ccgccttagc tttctcgtga cctagtcgtc ctcgtctttt cttcttcttc ttctataaaa
     7801 caatacccaa agagctcttc ttcttcacaa ttcagatttc aatttctcaa aatcttaaaa
     7861 actttctctc aattctctct accgtgatca aggtaaattt ctgtgttcct tattctctca
     7921 aaatcttcga ttttgttttc gttcgatccc aatttcgtat atgttctttg gtttagattc
     7981 tgttaatctt agatcgaaga cgattttctg ggtttgatcg ttagatatca tcttaattct
     8041 cgattagggt ttcatagata tcatccgatt tgttcaaata atttgagttt tgtcgaataa
     8101 ttactcttcg atttgtgatt tctatctaga tctggtgtta gtttctagtt tgtgcgatcg
     8161 aatttgtaga ttaatctgag tttttctgat taacagctcg agtgcgggcc ccccctcgag
     8221 gcgcgccaag ctatcaaaca agtttgtaca aaaaagcagg ctaccatatg gccagcagcc
     8281 cccccaagaa gaagcgcaag gtcgacaaga agtactccat cggcctggac atcggcacga
     8341 acagcgtggg ctgggccgtg atcaccgacg agtacaaggt gccgtccaag aagttcaagg
     8401 tgctgggcaa caccgaccgc cacagcatca agaagaacct gatcggcgcc ctgctgttcg
     8461 actcgggcga gaccgccgag gcgacgcgcc tgaagcgcac cgcgcgccgc cgctacacgc
     8521 gccgcaagaa ccgcatctgc tacctgcagg agatcttcag caacgagatg gccaaggtgg
     8581 acgactcgtt cttccaccgc ctggaggagt ccttcctggt ggaggaggac aagaagcacg
     8641 agcgccaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccga
     8701 cgatctacca cctgcgcaag aagctggtgg acagcaccga caaggcggac ctgcgcctga
     8761 tctacctggc cctggcgcac atgatcaagt tccgcggcca cttcctgatc gagggcgacc
     8821 tgaaccccga caactcggac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     8881 agctgttcga ggagaacccg atcaacgcct cgggcgtgga cgccaaggcg atcctgtccg
     8941 cgcgcctgtc caagagccgc cgcttggaga acctgatcgc ccagctgccc ggcgagaaga
     9001 agaacggcct gttcggcaac ctgatcgcgc tgagcctggg cctgacgccg aacttcaagt
     9061 cgaacttcga cctggccgag gacgcgaagc tgcagctgag caaggacacc tacgacgacg
     9121 acctggacaa cctgctggcc cagatcggcg accagtacgc ggacctgttc ctggccgcga
     9181 agaacctgtc ggacgccatc ctgctgtccg acatcctgcg cgtgaacacc gagatcacga
     9241 aggcccccct gtcggcgtcc atgatcaagc gctacgacga gcaccaccag gacctgaccc
     9301 tgctgaaggc gctggtgcgc cagcagctgc cggagaagta caaggagatc ttcttcgacc
     9361 agtccaagaa cggctacgcc ggctacatcg acggcggcgc cagccaggag gagttctaca
     9421 agttcatcaa gcccatcctg gagaagatgg acggcacgga ggagctgctg gtgaagctga
     9481 accgcgagga cctgctgcgc aagcagcgca ccttcgacaa cggcagcatc ccccaccaga
     9541 tccacctggg cgagctgcac gccatcctgc gccgccagga ggacttctac ccgttcctga
     9601 aggacaaccg cgagaagatc gagaagatcc tgacgttccg catcccctac tacgtgggcc
     9661 cgctggctcg cggcaactcc cgcttcgcgt ggatgacccg caagagcgag gagaccatca
     9721 cgccctggaa cttcgaggag gtggtggaca agggcgccag cgcgcagtcg ttcatcgagc
     9781 gcatgaccaa cttcgacaag aacctgccca acgagaaggt gctgccgaag cactccctgc
     9841 tgtacgagta cttcaccgtg tacaacgagc tgacgaaggt gaagtacgtg accgagggca
     9901 tgcgcaagcc cgccttcctg agcggcgagc agaagaaggc gatcgtggac ctgctgttca
     9961 agaccaaccg caaggtgacg gtgaagcagc tgaaggagga ctacttcaag aagatcgagt
    10021 gcttcgacag cgtggagatc tcgggcgtgg aggaccgctt caacgccagc ctgggcacct
    10081 accacgacct gctgaagatc atcaaggaca aggacttcct ggacaacgag gagaacgagg
    10141 acatcctgga ggacatcgtg ctgaccctga cgctgttcga ggaccgcgag atgatcgagg
    10201 agcgcctgaa gacgtacgcc cacctgttcg acgacaaggt gatgaagcag ctgaagcgcc
    10261 gccgctacac cggctggggc cgcctgagcc gcaagctgat caacggcatc cgcgacaagc
    10321 agtcgggcaa gaccatcctg gacttcctga agtccgacgg cttcgcgaac cgcaacttca
    10381 tgcagctgat ccacgacgac tcgctgacct tcaaggagga catccagaag gcccaggtgt
    10441 cgggccaggg cgactccctg cacgagcaca tcgccaacct ggcgggctcc cccgcgatca
    10501 agaagggcat cctgcagacc gtgaaggtgg tggacgagct ggtgaaggtg atgggccgcc
    10561 acaagccgga gaacatcgtg atcgagatgg cccgcgagaa ccagaccacg cagaagggcc
    10621 agaagaacag ccgcgagcgc atgaagcgca tcgaggaggg catcaaggag ctgggctcgc
    10681 agatcctgaa ggagcacccc gtggagaaca cccagctgca gaacgagaag ctgtacctgt
    10741 actacctgca gaacggccgc gacatgtacg tggaccagga gctggacatc aaccgcctgt
    10801 ccgactacga cgtggaccac atcgtgcccc agagcttcct gaaggacgac tcgatcgaca
    10861 acaaggtgct gacccgcagc gacaagaacc gcggcaagag cgacaacgtg ccgtcggagg
    10921 aggtggtgaa gaagatgaag aactactggc gccagctgct gaacgccaag ctgatcacgc
    10981 agcgcaagtt cgacaacctg accaaggccg agcgcggcgg cctgtcggag ctggacaagg
    11041 cgggcttcat caagcgccag ctggtggaga cccgccagat cacgaagcac gtggcgcaga
    11101 tcctggactc ccgcatgaac acgaagtacg acgagaacga caagctgatc cgcgaggtga
    11161 aggtgatcac cctgaagtcc aagctggtca gcgacttccg caaggacttc cagttctaca
    11221 aggtgcgcga gatcaacaac taccaccacg cccacgacgc gtacctgaac gccgtggtgg
    11281 gcaccgcgct gatcaagaag taccccaagc tggagagcga gttcgtgtac ggcgactaca
    11341 aggtgtacga cgtgcgcaag atgatcgcca agtcggagca ggagatcggc aaggccaccg
    11401 cgaagtactt cttctactcc aacatcatga acttcttcaa gaccgagatc acgctggcca
    11461 acggcgagat ccgcaagcgc cccctgatcg agaccaacgg cgagacgggc gagatcgtgt
    11521 gggacaaggg ccgcgacttc gcgaccgtgc gcaaggtgct gagcatgccc caggtgaaca
    11581 tcgtgaagaa gaccgaggtg cagacgggcg gcttctccaa ggagagcatc ctgccgaagc
    11641 gcaactcgga caagctgatc gcccgcaaga aggactggga ccccaagaag tacggcggct
    11701 tcgactcccc gaccgtggcc tacagcgtgc tggtggtggc gaaggtggag aagggcaagt
    11761 ccaagaagct gaagagcgtg aaggagctgc tgggcatcac catcatggag cgcagctcgt
    11821 tcgagaagaa ccccatcgac ttcctggagg cgaagggcta caaggaggtg aagaaggacc
    11881 tgatcatcaa gctgccgaag tactcgctgt tcgagctgga gaacggccgc aagcgcatgc
    11941 tggcctccgc gggcgagctg cagaagggca acgagctggc cctgcccagc aagtacgtga
    12001 acttcctgta cctggcgtcc cactacgaga agctgaaggg ctcgccggag gacaacgagc
    12061 agaagcagct gttcgtggag cagcacaagc actacctgga cgagatcatc gagcagatct
    12121 cggagttctc caagcgcgtg atcctggccg acgcgaacct ggacaaggtg ctgagcgcct
    12181 acaacaagca ccgcgacaag cccatccgcg agcaggcgga gaacatcatc cacctgttca
    12241 ccctgacgaa cctgggcgcc ccggccgcgt tcaagtactt cgacaccacg atcgaccgca
    12301 agcgctacac cagcacgaag gaggtgctgg acgcgaccct gatccaccag agcatcaccg
    12361 gcctgtacga gacgcgcatc gacctgtcgc agctgggcgg cgactaatgt acccagcttt
    12421 cttgtacaaa gtggttcgat aattcttaat taactagttc tagagcggcc gccaccgcgg
    12481 tggagctcga atttccccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
    12541 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
    12601 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
    12661 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
    12721 tcgcgcgcgg tgtcatctat gttactagat cgggaattcg taatcatgtc atagctgttt
    12781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
    12841 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
    12901 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
    12961 gggagaggcg gtttgcgtat tggctagagc agcttgccaa catggtggag cacgacactc
    13021 tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt
    13081 ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact
    13141 tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag
    13201 gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
    13261 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
    13321 gtgaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca
    13381 gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
    13441 ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc
    13501 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt
    13561 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
    13621 acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa
    13681 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
    13741 acacgctgaa atcaccagtc tctctctaca aatctatctc tctcgagctt tcgcagatcc
    13801 ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
    13861 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
    13921 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
    13981 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
    14041 gaagtgcttg acattgggga gtttagcgag agcctgacct attgcatctc ccgccgttca
    14101 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct acaaccggtc
    14161 gcggaggcta tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
    14221 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
    14281 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
    14341 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
    14401 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
    14461 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
    14521 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
    14581 cttgcaggat cgccacgact ccgggcgtat atgctccgca ttggtcttga ccaactctat
    14641 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
    14701 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
    14761 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
    14821 cgtccgaggg caaagaaata g
//
