LOCUS       sequence-491208-       12690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A promoterless construct designed for cloning various Pol III 
            promoters or serving as a control in evaluating Pol III promoters 
            for CRISPR-Cas9 editing of mEGFP in Nicotiana benthamiana and poplar
            reporter lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12690)
  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 12690)
  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: p305N
FEATURES             Location/Qualifiers
     source          1..12690
                     /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        6284..6359
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             8043..12146
                     /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             8043..12146
                     /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             8043..12146
                     /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      12186..12434
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    12453..12477
                     /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 gatgaaggta ccgcatgccc gaaggctacg tccgttttag agctagaaat
     6301 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     6361 ttttttacta gtggagctca acttttgatt cgctatttgc agtgcacctg tggcgttcat
     6421 cacatctttt gtgacactgt ttgcactggt cattgctatt acaaaggacc ttcctgatgt
     6481 tgaaggagat cgaaagtaag taactgcacg cataaccatt ttctttccgc tctttggctc
     6541 aatccatttg acagtcaaag acaatgttta accagctccg tttgatatat tgtctttatg
     6601 tgtttgttca agcatgttta gttaatcatg cctttgattg atcttgaata ggttccaaat
     6661 atcaaccctg gcaacaaaac ttggagtgag aaacattgca ttcctcggtt ctggacttct
     6721 gctagtaaat tatgtttcag ccatatcact agctttctac atgcctcagg tgaattcatc
     6781 tatttccgtc ttaactattt cggttaatta aagcacgaac accattactg catgtagaag
     6841 cttgataaac tatcgccacc aatttatttt tgttgcgata ttgttacttt cctcagtatg
     6901 cagctttgaa aagaccaacc ctcttatcct ttaacaatga acaggttttt agaggtagct
     6961 tgatgattcc tgcacatgtg atcttggctt caggcttaat tttccaggta aagcattatg
     7021 agatactctt atatctctta catacttttg agataatgca caagaacttc ataactatat
     7081 gctttagttt ctgcatttga cactgccaaa ttcattaatc tctaatatct ttgttgttga
     7141 tctttggtag acatgggtac tagaaaaagc aaactacacc aaggtaaaat acttttgtac
     7201 aaacataaac tcgttatcac ggaacatcaa tggagtgtat atctaacgga gtgtagaaac
     7261 atttgattat tgcaggaagc tatctcagga tattatcggt ttatatggaa tctcttctac
     7321 gcagagtatc tgttattccc cttcctctag ctttcaattt catggtgagg atatgcagtt
     7381 ttctttgtat atcattcttc ttcttctttg tagcttggag tcaaaatcgg ttccttcatg
     7441 tacatacatc aaggatatgt ccttctgaat ttttatatct tgcaataaaa atgcttgtac
     7501 caattgaaac accagctttt tgagttctat gatcactgac ttggttctaa ccaaaaaaaa
     7561 aaaaatgttt aatttacata tctaaaagta ggtttaggga aacctaaaca gtaaaatatt
     7621 tgtatattat tcgaatttca ctcatcataa aaacttaaat tgcaccataa aattttgttt
     7681 tactattaat gatgtaattt gtgtaactta agataaaaat aatattccgt aagttaaccg
     7741 gctaaaacca cgtataaacc agggaacctg ttaaaccggt tctttactgg ataaagaaat
     7801 gaaagcccat gtagacagct ccattagagc ccaaacccta aatttctcat ctatataaaa
     7861 ggagtgacat tagggttttt gttcgtcctc ttaaagcttc tcgttttctc tgccgtctct
     7921 ctcattcgcg cgacgcaaac gatcttcagg tgatcttctt tctccaaatc ctctctcata
     7981 actctgattt cgtacttgtg tatttgagct cacgctctgt ttctctcacc acagcagcta
     8041 gcatggacaa gaagtactcc attgggctcg atatcggcac aaacagcgtc ggctgggccg
     8101 tcattacgga cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc
     8161 gccacagcat aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg
     8221 aagccacgcg gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct
     8281 gctacctgca ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata
     8341 ggctggagga gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg
     8401 gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga
     8461 agaagcttgt agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc
     8521 atatgatcaa atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg
     8581 atgtcgacaa actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc
     8641 cgatcaacgc atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc
     8701 ggcggctcga aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta
     8761 atcttatcgc cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg
     8821 aagatgccaa gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg
     8881 cccagatcgg cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca
     8941 ttctgctgag tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta
     9001 gtatgatcaa gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca
     9061 gacagcaact gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg
     9121 ccggatacat tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct
     9181 tggaaaaaat ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc
     9241 gcaaacagcg cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc
     9301 acgctatcct caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga
     9361 ttgagaaaat cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt
     9421 ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg
     9481 aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata
     9541 aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag
     9601 tttataacga gctgaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc
     9661 tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta
     9721 ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa
     9781 tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa
     9841 tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg
     9901 tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg
     9961 ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg
    10021 ggcggctgtc aagaaaactg atcaatggca tccgagacaa gcagagtgga aagacaatcc
    10081 tggattttct taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg
    10141 actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc
    10201 ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga
    10261 ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg
    10321 ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa
    10381 ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc
    10441 cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca
    10501 gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc
    10561 atatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat
    10621 ccgataaaaa tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga
    10681 aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc
    10741 tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc
    10801 agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga
    10861 acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt
    10921 ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca
    10981 attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa
    11041 aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga
    11101 aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca
    11161 gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc
    11221 gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt
    11281 tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag
    11341 tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga
    11401 tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg
    11461 cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg
    11521 tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg
    11581 actttctcga agcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca
    11641 agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc
    11701 tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca
    11761 gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg
    11821 aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgagttc tccaaaagag
    11881 tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata
    11941 agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg
    12001 cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa
    12061 aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa
    12121 tcgacctctc tcagctcggt ggagacggaa caagaaagag aagaaaggtg tgacggatcc
    12181 tcgaatatga agatgaagat gaaatatttg gtgtgtcaaa taaaaagctt gtgtgcttaa
    12241 gtttgtgttt ttttcttggc ttgttgtgtt atgaatttgt ggctttttct aatatcaaat
    12301 gaatgtaaga tctcattata atgaataaac aaatgtttct ataatccatt gtgaatgttt
    12361 tgttggatct cttctgcagc atataactac tgtatgtgct atggtatgga ctatggaata
    12421 tgattaaaga taaggtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta
    12481 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
    12541 gttcgtccat ttgtatgtgc atgccaacca caggagatcc aaaaggatct aggtgaagat
    12601 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    12661 agaccccgta gaaaagatca aaggatcttc
//
