LOCUS       sequence-491189-       13042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant CRISPR editing with the experimentally validated Medicago 
            truncatula MtU6.6 promoter (352 bp).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13042)
  AUTHORS   Deguchi M, Sinclair KM, Patel A, Coile M, Ortega MA, Bewg WP, Tsai 
            CJ
  TITLE     A compendium of nonredundant short polymerase III promoters for 
            CRISPR applications.
  JOURNAL   Plant Physiol. 2025 Jul 3;198(3):kiaf294. doi: 
            10.1093/plphys/kiaf294.
  PUBMED    40673482
REFERENCE   2  (bases 1 to 13042)
  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 
            Physiol."; date: "2025-07-3"; volume: "198(3):kiaf294. doi"; pages: 
            " 10.1093/plphys/kiaf294"
COMMENT     Sequence Label: p304N
FEATURES             Location/Qualifiers
     source          1..13042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(829..1977)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MAQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(2276..3070)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3282..3913
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    3993..4017
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(4532..5326)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4643..4662)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5253..5272
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_RNA        6636..6711
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             8395..12498
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8395..12498
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8395..12498
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     terminator      12538..12786
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    12805..12829
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag aggatctcag
      781 taaagcgctg gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca
      841 atgcaccagg tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc
      901 gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat
      961 gaggcggaag gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa
     1021 catccgtcgg gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg
     1081 gtcgccagca aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt
     1141 cttgccacgg tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac
     1201 gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt
     1261 cgtgtaatac cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa
     1321 ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc aacacctgct gccacaccag
     1381 ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac
     1441 gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa
     1501 cagggcagag cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat
     1561 atcgaacaag gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc
     1621 ttgcttggcc tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt
     1681 cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc
     1741 gagacgacgc gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg
     1801 cacgctgtcg cgctcgatct tggccgtagc ttgctgggcc atcgagccga cggactggaa
     1861 ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga
     1921 aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca
     1981 ccctccttgc gggattgccc cgactcacgc cggggcaatg tgcccttatt cctgatttga
     2041 tccgcctggt gccttggtgt ccagataatc caccttatcg gcaatgaagt cggtcccgta
     2101 gaccgtctgg ccgtccttct cgtacttggt attccgaatc ttgccctgca cgaataccag
     2161 cgaccccttg cccaaatact tgccgtgggc ctcggcctga gagccaaaac acttgatgcg
     2221 gaagaagtcg gtgcgctcct gcttgtcgcc agcatcgttg cgccacatct aggtactaaa
     2281 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta
     2341 agtcaaaaaa tagctcgaca tactgttctt ccccaatatc ctccctgatc gaccggacgc
     2401 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac
     2461 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa
     2521 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg
     2581 gggtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat
     2641 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg
     2701 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat
     2761 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc
     2821 catcatgccg ttcaaggtgc aggacctttg gaacaggcag ctttccttcc agccatagca
     2881 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt
     2941 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt
     3001 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca
     3061 ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc
     3121 taattataac aagacgaact ccaattcact gttccttgca ttctaaaact ttaaatacca
     3181 gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
     3241 gattttgaaa ccacaattat gggtgatgct gccaacgcga gagcgggccg ggagggttcg
     3301 agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc agccctggtt
     3361 aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt agcggtggcc
     3421 gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca gcccctcaaa
     3481 tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca aggatcgcgc
     3541 ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac ttatccccag
     3601 gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc cgatttgcga
     3661 ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg tcaacgccgc
     3721 gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag tgagggccaa
     3781 gttttccgcg aggtatccac aacgccggcg gctgcggtgt ctcgcacacg gcttcgacgg
     3841 cgtttctggc gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag
     3901 cccggtgagc gtctctagtg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
     3961 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
     4021 ttgacgattt aaatggtagg cgcggcttaa ttaagcgctt aattaagcgc ttaattaagc
     4081 gcttaataag ccgcgccgta ccccaatagt ctcgacagac acatagcacc taacattatt
     4141 gcttaaattt actatttttt tactatatta tactcaaccc aatgagcata aagactgtaa
     4201 aatctcaaat tcctgagaag catatttatc gatcccacag acttgatagt tccataatcc
     4261 atacgctgca gccaaattgc tagtgtgttg aacatttaac acgtagagaa ctagaaaaga
     4321 tataaaacta agattgatat ccaaaataga cgagaacaat aagcaaaaac tcttagtttt
     4381 gaaataaatc aacaatcccg agggttgtca catacatcaa aaacgaaaat ccatatagca
     4441 aaaaaaactc taaattaccg ttcgacaaaa agagaaaact gataggacat ttgctaaaca
     4501 ttaaaatcaa aatttggatc aagccgaatt ctcagaagaa ctcgtcaaga aggcgataga
     4561 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     4621 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     4681 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     4741 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc atcgccgtcg ggcatgcgcg
     4801 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     4861 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     4921 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     4981 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     5041 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     5101 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg
     5161 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     5221 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     5281 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatctcg agacgtaccg
     5341 cgcgatgcat tcgaagatcg atccttcgcc tggaggagag aaatcagtgg cgctgcggct
     5401 tttagggttt ctttgttgat ggaatgagag tgtaagctct gccagtgcca ctttattagg
     5461 gttttacaag cccttttctt cgtaattggg cctgacattt tgtgccactt gggcctttag
     5521 agatgaaaat gtatattggg cttaagttga cttgaaggat aaattagttt aggatattac
     5581 gttttttatg agaattggtg tgtcggatac atgtatatga tgcattcaaa tatatgtatt
     5641 ctagatacat ttaagtttag atacaatcta aaatgtgtct ttaattacag gactgtaact
     5701 aaaatactta atgtaagaag aatattactc ctttaatagc ttttgagtat atctagtcta
     5761 acatctttta aaaaagtcta atttctttca tttatttttc gagcaatagc aaagtgcata
     5821 attatttttt tcttctagaa attcagattt gtttctctaa attttgagat tcttttctca
     5881 attttgtatg tctagagaac aatgtgtatt tttcactcta gttggttgtt actttgttga
     5941 atgttctgat aaaagtatat tgttatttct gaagtagata taaaccttca tttggaaatt
     6001 atacataaat caaaatcgtt aattatctag atcaagatat atgccctttt cctaatgtat
     6061 ttgatacatg cacctaattt cactagatgt atcttttcta ttttttaaat tatgaatagt
     6121 taattttttc catatgtgta tttgatacat acttcatgac tttaaaaaat taattatata
     6181 ccagatatat gtatttaaaa tttgttatgt atttaaagta tgtatatgat tattcgatat
     6241 taatctcttc gatgaaccta ggatgcctat cttatatgat caatgaggca tttaattggg
     6301 tgcatatgat ggtgaaaaaa ggtgcagctc ctggcttggg aatgatgact catgtggaat
     6361 ttggtcttaa atttatcaca tccttttggg atgtgatgat tgtatcactt gttcattttg
     6421 caaagacaag gtgcactgct acaaactttg gtttaatctg aaataaaaca aaactcactg
     6481 agaggaagat gcatcccagt aggtgaaagt cgagaaggat ttgcatgtta ctattacact
     6541 tgctttttag tcccacatcg tctgaaacat aaaatatttc agcgtttaaa tacttcaagc
     6601 gaaccagtag gcttgcatgc ccgaaggcta cgtccgtttt agagctagaa atagcaagtt
     6661 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttttac
     6721 tagtggagct caacttttga ttcgctattt gcagtgcacc tgtggcgttc atcacatctt
     6781 ttgtgacact gtttgcactg gtcattgcta ttacaaagga ccttcctgat gttgaaggag
     6841 atcgaaagta agtaactgca cgcataacca ttttctttcc gctctttggc tcaatccatt
     6901 tgacagtcaa agacaatgtt taaccagctc cgtttgatat attgtcttta tgtgtttgtt
     6961 caagcatgtt tagttaatca tgcctttgat tgatcttgaa taggttccaa atatcaaccc
     7021 tggcaacaaa acttggagtg agaaacattg cattcctcgg ttctggactt ctgctagtaa
     7081 attatgtttc agccatatca ctagctttct acatgcctca ggtgaattca tctatttccg
     7141 tcttaactat ttcggttaat taaagcacga acaccattac tgcatgtaga agcttgataa
     7201 actatcgcca ccaatttatt tttgttgcga tattgttact ttcctcagta tgcagctttg
     7261 aaaagaccaa ccctcttatc ctttaacaat gaacaggttt ttagaggtag cttgatgatt
     7321 cctgcacatg tgatcttggc ttcaggctta attttccagg taaagcatta tgagatactc
     7381 ttatatctct tacatacttt tgagataatg cacaagaact tcataactat atgctttagt
     7441 ttctgcattt gacactgcca aattcattaa tctctaatat ctttgttgtt gatctttggt
     7501 agacatgggt actagaaaaa gcaaactaca ccaaggtaaa atacttttgt acaaacataa
     7561 actcgttatc acggaacatc aatggagtgt atatctaacg gagtgtagaa acatttgatt
     7621 attgcaggaa gctatctcag gatattatcg gtttatatgg aatctcttct acgcagagta
     7681 tctgttattc cccttcctct agctttcaat ttcatggtga ggatatgcag ttttctttgt
     7741 atatcattct tcttcttctt tgtagcttgg agtcaaaatc ggttccttca tgtacataca
     7801 tcaaggatat gtccttctga atttttatat cttgcaataa aaatgcttgt accaattgaa
     7861 acaccagctt tttgagttct atgatcactg acttggttct aaccaaaaaa aaaaaaatgt
     7921 ttaatttaca tatctaaaag taggtttagg gaaacctaaa cagtaaaata tttgtatatt
     7981 attcgaattt cactcatcat aaaaacttaa attgcaccat aaaattttgt tttactatta
     8041 atgatgtaat ttgtgtaact taagataaaa ataatattcc gtaagttaac cggctaaaac
     8101 cacgtataaa ccagggaacc tgttaaaccg gttctttact ggataaagaa atgaaagccc
     8161 atgtagacag ctccattaga gcccaaaccc taaatttctc atctatataa aaggagtgac
     8221 attagggttt ttgttcgtcc tcttaaagct tctcgttttc tctgccgtct ctctcattcg
     8281 cgcgacgcaa acgatcttca ggtgatcttc tttctccaaa tcctctctca taactctgat
     8341 ttcgtacttg tgtatttgag ctcacgctct gtttctctca ccacagcagc tagcatggac
     8401 aagaagtact ccattgggct cgatatcggc acaaacagcg tcggctgggc cgtcattacg
     8461 gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc
     8521 ataaagaaga acctcattgg cgccctcctg ttcgactccg gggagacggc cgaagccacg
     8581 cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg
     8641 caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag
     8701 gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc
     8761 gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctt
     8821 gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcatatgatc
     8881 aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac
     8941 aaactcttta tccaactggt tcagacttac aatcagcttt tcgaagagaa cccgatcaac
     9001 gcatccggag ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc
     9061 gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc
     9121 gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc
     9181 aagcttcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc
     9241 ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg
     9301 agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc
     9361 aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa
     9421 ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac
     9481 attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa
     9541 atggacggca ccgaggagct gctggtaaag cttaacagag aagatctgtt gcgcaaacag
     9601 cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc
     9661 ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa
     9721 atcctcacat ttcggatacc ctactatgta ggccccctcg cccggggaaa ttccagattc
     9781 gcgtggatga ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg
     9841 gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg
     9901 cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac
     9961 gagctgacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga
    10021 gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa
    10081 cagctcaaag aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga
    10141 gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa
    10201 gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc
    10261 cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc
    10321 ttcgacgaca aagtcatgaa acagctcaag aggcgccgat atacaggatg ggggcggctg
    10381 tcaagaaaac tgatcaatgg catccgagac aagcagagtg gaaagacaat cctggatttt
    10441 cttaagtccg atggatttgc caaccggaac ttcatgcagt tgatccatga tgactctctc
    10501 acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag
    10561 cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag
    10621 gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag
    10681 atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag
    10741 aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa
    10801 aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg
    10861 tacgtggatc aggaactgga catcaatcgg ctctccgact acgacgtgga tcatatcgtg
    10921 ccccagtctt ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa
    10981 aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat
    11041 tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag
    11101 gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt
    11161 gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag
    11221 tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg
    11281 gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac
    11341 catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc
    11401 aagcttgaat ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc
    11461 gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt
    11521 atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt
    11581 atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca
    11641 gtccggaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc
    11701 ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc
    11761 aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt
    11821 gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa
    11881 ctgctgggca tcacaatcat ggagcgatca agcttcgaaa aaaaccccat cgactttctc
    11941 gaagcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagcttcc caagtactct
    12001 ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa
    12061 ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat
    12121 gaaaagctca aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac
    12181 aaacactacc ttgatgagat catcgagcaa ataagcgagt tctccaaaag agtgatcctc
    12241 gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc
    12301 agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca
    12361 gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc
    12421 ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc
    12481 tctcagctcg gtggagacgg aacaagaaag agaagaaagg tgtgacggat cctcgaatat
    12541 gaagatgaag atgaaatatt tggtgtgtca aataaaaagc ttgtgtgctt aagtttgtgt
    12601 ttttttcttg gcttgttgtg ttatgaattt gtggcttttt ctaatatcaa atgaatgtaa
    12661 gatctcatta taatgaataa acaaatgttt ctataatcca ttgtgaatgt tttgttggat
    12721 ctcttctgca gcatataact actgtatgtg ctatggtatg gactatggaa tatgattaaa
    12781 gataaggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
    12841 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
    12901 atttgtatgt gcatgccaac cacaggagat ccaaaaggat ctaggtgaag atcctttttg
    12961 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
    13021 tagaaaagat caaaggatct tc
//
