LOCUS       sequence-494268-       13958 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  Evaluation of AtU6.1m2 promoter (Pol III promoter) sequence 
            variation for CRISPR-Cas9 editing of mEGFP in a Nicotiana 
            benthamiana reporter line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13958)
  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 13958)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 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-5
FEATURES             Location/Qualifiers
     source          1..13958
                     /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(3475..4269)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4481..5112
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5192..5216
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(5731..6525)
                     /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(5842..5861)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6452..6471
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_RNA        7552..7627
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             9311..13414
                     /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             9311..13414
                     /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             9311..13414
                     /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      13454..13702
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     misc_feature    13721..13745
                     /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 actttgtttt 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 agggaaggtg
     2281 cgaacaagtc cctgatatga gatcatgttt gtcatctgga gccatagaac agggttcatc
     2341 atgagtcatc aacttacctt cgccgacagt gaattcagca gtaagcgccg tcagaccaga
     2401 aaagagattt tcttgtcccg catggagcag attctgccat ggcaaaacat ggtggaagtc
     2461 atcgagccgt tttaccccaa ggctggtaat ggccggcgac cttatccgct ggaaaccatg
     2521 ctacgcattc actgcatgca gcattggtac aacctgagcg atggcgcgat ggaagatgct
     2581 ctgtacgaaa tcgcctccat gcgtctgttt gcccggttat ccctggatag cgccttgccg
     2641 gaccgcacca ccatcatgaa tttccgccac ctgctggagc agcatcaact ggcccgccaa
     2701 ttgttcaaga ccatcaatcg ctggctggcc gaagcaggcg tcatgatgac tcaaggcacc
     2761 ttggtcgatg ccaccatcat tgaggcaccc agctcgacca agaacaaaga gcagcaacgc
     2821 gatccggaga tgcatcagac caagaaaggc aatcagtggc actttggcat gaaggcccac
     2881 attggtgtcg atgccaagag tggcctgacc cacagcctgg tcaccaccgc ggccaacgag
     2941 catgacctca atcagctggg taatctgctg catggagagg agcaatttgt ctcagccgat
     3001 gccggctacc aaggggcgcc acagcgcgag gagctggccg aggtggatgt ggactggctg
     3061 atcgccgagc gccccggcaa ggtaagaacc ttgaaacagc atccacgcaa gaacaaaacg
     3121 gccatcaaca tcgaatacat gaaagccagc atccgggcca gggtggagca cccatttcgc
     3181 atcatcaagc gacagttcgg cttcgtgaaa gccagataca aggggttgct gaaaaacgat
     3241 aaccaactgg cgatgttatt cacgctggcc aacctgtttc gggcggacca aatgatacgt
     3301 cagtgggaga gatctcacta aaaactgggg ataacgcctt aaatggcgaa gaaacggtct
     3361 aaataggctg attcaaggca tttacgggag aaaaaatcgg ctcaaacatg aagaaatgaa
     3421 atgactgagt cagccgagaa gaatttcccc gcttattcgc accttcccta ggtactaaaa
     3481 caattcatcc agtaaaatat aatattttat tttctcccaa tcaggcttga tccccagtaa
     3541 gtcaaaaaat agctcgacat actgttcttc cccaatatcc tccctgatcg accggacgca
     3601 gaaggcaatg tcataccact tgtccgccct gccgcttctc ccaagatcaa taaagccact
     3661 tactttgcca tctttcacaa agatgttgct gtctcccagg tcgccgtggg aaaagacaag
     3721 ttcctcttcg ggcttttccg tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg
     3781 ggtgtcttct tcccagtttt cgcaatccac atcggccaga tcgttattca gtaagtaatc
     3841 caattcggct aagcggctgt ctaagctatt cgtataggga caatccgata tgtcgatgga
     3901 gtgaaagagc ctgatgcact ccgcatacag ctcgataatc ttttcagggc tttgttcatc
     3961 ttcatactct tccgagcaaa ggacgccatc ggcctcactc atgagcagat tgctccagcc
     4021 atcatgccgt tcaaggtgca ggacctttgg aacaggcagc tttccttcca gccatagcat
     4081 catgtccttt tcccgttcca catcataggt ggtcccttta taccggctgt ccgtcatttt
     4141 taaatatagg ttttcatttt ctcccaccag cttatatacc ttagcaggag acattccttc
     4201 cgtatctttt acgcagcggt atttttcgat cagttttttc aattccggtg atattctcat
     4261 tttagccatt tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct
     4321 aattataaca agacgaactc caattcactg ttccttgcat tctaaaactt taaataccag
     4381 aaaacagctt tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg
     4441 attttgaaac cacaattatg ggtgatgctg ccaacgcgag agcgggccgg gagggttcga
     4501 gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta
     4561 aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg
     4621 aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat
     4681 gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc
     4741 cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg
     4801 cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag
     4861 gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg
     4921 ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag
     4981 ttttccgcga ggtatccaca acgccggcgg ctgcggtgtc tcgcacacgg cttcgacggc
     5041 gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc
     5101 ccggtgagcg tctctagtgg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
     5161 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
     5221 tgacgattta aatggtaggc gcggcttaat taagcgctta attaagcgct taattaagcg
     5281 cttaataagc cgcgccgtac cccaatagtc tcgacagaca catagcacct aacattattg
     5341 cttaaattta ctattttttt actatattat actcaaccca atgagcataa agactgtaaa
     5401 atctcaaatt cctgagaagc atatttatcg atcccacaga cttgatagtt ccataatcca
     5461 tacgctgcag ccaaattgct agtgtgttga acatttaaca cgtagagaac tagaaaagat
     5521 ataaaactaa gattgatatc caaaatagac gagaacaata agcaaaaact cttagttttg
     5581 aaataaatca acaatcccga gggttgtcac atacatcaaa aacgaaaatc catatagcaa
     5641 aaaaaactct aaattaccgt tcgacaaaaa gagaaaactg ataggacatt tgctaaacat
     5701 taaaatcaaa atttggatca agccgaattc tcagaagaac tcgtcaagaa ggcgatagaa
     5761 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     5821 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     5881 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     5941 attcggcaag caggcatcgc catgggtcac gacgagatca tcgccgtcgg gcatgcgcgc
     6001 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     6061 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     6121 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     6181 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     6241 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     6301 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     6361 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     6421 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     6481 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatctcga gacgtaccgc
     6541 gcgatgcatt cgaagatcga tccttcgcct ggaggagaga aatcagtggc gctgcggctt
     6601 ttagggtttc tttgttgatg gaatgagagt gtaagctctg ccagtgccac tttattaggg
     6661 ttttacaagc ccttttcttc gtaattgggc ctgacatttt gtgccacttg ggcctttaga
     6721 gatgaaaatg tatattgggc ttaagttgac ttgaaggata aattagttta ggatattacg
     6781 ttttttatga gaattggtgt gtcggataca tgtatatgat gcattcaaat atatgtattc
     6841 tagatacatt taagtttaga tacaatctaa aatgtgtctt taattacagg actgtaacta
     6901 aaatacttaa tgtaagaaga atattactcc tttaatagct tttgagtata tctagtctaa
     6961 catcttttaa aaaagtctaa tttctttcat ttatttttcg agcaatagca aagtgcataa
     7021 ttattttttt cttctagaaa ttcagatttg tttctctaaa ttttgagatt cttttctcaa
     7081 ttttgtatgt ctagagaaca atgtgtattt ttcactctag ttggttgtta ctttgttgaa
     7141 tgttctgata aaagtatatt gttatttctg aagtagatat aaaccttcat ttggaaatta
     7201 tacataaatc aaaatcgtta attatctaga tcaagatata tgcccttttc ctaatgtatt
     7261 tgatacatgc acctaatttc actagatgta tcttttctat tttttaaatt atgaatagtt
     7321 aattttttcc atatgtgtat ttgatacata cttcatgact ttaaaaaatt aattatatac
     7381 cagatatatg tatttaaaat ttgttatgta tttaaagtat gtatatgatt attcgatatt
     7441 aatctcttcg atgaaggcct caaaagtccc acatcgctta ggtaaagaaa gcagctgagt
     7501 ttatatatgg ttagagacga agtagtgatt gcatgcccga aggctacgtc cgttttagag
     7561 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     7621 tcggtgcttt ttttactagt ggagctcaac ttttgattcg ctatttgcag tgcacctgtg
     7681 gcgttcatca catcttttgt gacactgttt gcactggtca ttgctattac aaaggacctt
     7741 cctgatgttg aaggagatcg aaagtaagta actgcacgca taaccatttt ctttccgctc
     7801 tttggctcaa tccatttgac agtcaaagac aatgtttaac cagctccgtt tgatatattg
     7861 tctttatgtg tttgttcaag catgtttagt taatcatgcc tttgattgat cttgaatagg
     7921 ttccaaatat caaccctggc aacaaaactt ggagtgagaa acattgcatt cctcggttct
     7981 ggacttctgc tagtaaatta tgtttcagcc atatcactag ctttctacat gcctcaggtg
     8041 aattcatcta tttccgtctt aactatttcg gttaattaaa gcacgaacac cattactgca
     8101 tgtagaagct tgataaacta tcgccaccaa tttatttttg ttgcgatatt gttactttcc
     8161 tcagtatgca gctttgaaaa gaccaaccct cttatccttt aacaatgaac aggtttttag
     8221 aggtagcttg atgattcctg cacatgtgat cttggcttca ggcttaattt tccaggtaaa
     8281 gcattatgag atactcttat atctcttaca tacttttgag ataatgcaca agaacttcat
     8341 aactatatgc tttagtttct gcatttgaca ctgccaaatt cattaatctc taatatcttt
     8401 gttgttgatc tttggtagac atgggtacta gaaaaagcaa actacaccaa ggtaaaatac
     8461 ttttgtacaa acataaactc gttatcacgg aacatcaatg gagtgtatat ctaacggagt
     8521 gtagaaacat ttgattattg caggaagcta tctcaggata ttatcggttt atatggaatc
     8581 tcttctacgc agagtatctg ttattcccct tcctctagct ttcaatttca tggtgaggat
     8641 atgcagtttt ctttgtatat cattcttctt cttctttgta gcttggagtc aaaatcggtt
     8701 ccttcatgta catacatcaa ggatatgtcc ttctgaattt ttatatcttg caataaaaat
     8761 gcttgtacca attgaaacac cagctttttg agttctatga tcactgactt ggttctaacc
     8821 aaaaaaaaaa aaatgtttaa tttacatatc taaaagtagg tttagggaaa cctaaacagt
     8881 aaaatatttg tatattattc gaatttcact catcataaaa acttaaattg caccataaaa
     8941 ttttgtttta ctattaatga tgtaatttgt gtaacttaag ataaaaataa tattccgtaa
     9001 gttaaccggc taaaaccacg tataaaccag ggaacctgtt aaaccggttc tttactggat
     9061 aaagaaatga aagcccatgt agacagctcc attagagccc aaaccctaaa tttctcatct
     9121 atataaaagg agtgacatta gggtttttgt tcgtcctctt aaagcttctc gttttctctg
     9181 ccgtctctct cattcgcgcg acgcaaacga tcttcaggtg atcttctttc tccaaatcct
     9241 ctctcataac tctgatttcg tacttgtgta tttgagctca cgctctgttt ctctcaccac
     9301 agcagctagc atggacaaga agtactccat tgggctcgat atcggcacaa acagcgtcgg
     9361 ctgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
     9421 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
     9481 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
     9541 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
     9601 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
     9661 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
     9721 tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc
     9781 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
     9841 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
     9901 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
     9961 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
    10021 gtttggtaat cttatcgccc tgtcactcgg gctgaccccc aactttaaat ctaacttcga
    10081 cctggccgaa gatgccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
    10141 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
    10201 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
    10261 gagcgctagt atgatcaagc gctatgatga gcaccaccaa gacttgactt tgctgaaggc
    10321 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
    10381 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
    10441 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
    10501 tctgttgcgc aaacagcgca ctttcgacaa tggaagcatc ccccaccaga ttcacctggg
    10561 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
    10621 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
    10681 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
    10741 cttcgaggaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
    10801 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
    10861 cttcacagtt tataacgagc tgaccaaggt caaatacgtc acagaaggga tgagaaagcc
    10921 agcattcctg tctggagagc agaagaaagc tatcgtggac ctcctcttca agacgaaccg
    10981 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
    11041 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
    11101 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
    11161 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
    11221 aacttacgct catctcttcg acgacaaagt catgaaacag ctcaagaggc gccgatatac
    11281 aggatggggg cggctgtcaa gaaaactgat caatggcatc cgagacaagc agagtggaaa
    11341 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca tgcagttgat
    11401 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
    11461 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
    11521 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
    11581 gaatatcgtt atcgagatgg cccgagagaa ccaaactacc cagaagggac agaagaacag
    11641 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
    11701 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
    11761 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
    11821 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
    11881 gacaagatcc gataaaaata gagggaagag tgataacgtc ccctcagaag aagttgtcaa
    11941 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
    12001 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggcttcat
    12061 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
    12121 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
    12181 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
    12241 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
    12301 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
    12361 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
    12421 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
    12481 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
    12541 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa
    12601 gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga
    12661 caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc
    12721 tacagtcgct tacagtgtac tggttgtggc caaagtggag aaagggaagt ctaaaaaact
    12781 caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa
    12841 ccccatcgac tttctcgaag cgaaaggata taaagaggtc aaaaaagacc tcatcattaa
    12901 gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc
    12961 gggcgagctg cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta
    13021 tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct
    13081 gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa gcgagttctc
    13141 caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca
    13201 cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa
    13261 cttgggcgcg cctgcagcct tcaagtactt cgacaccacc atagacagaa agcggtacac
    13321 ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga
    13381 aacaagaatc gacctctctc agctcggtgg agacggaaca agaaagagaa gaaaggtgtg
    13441 acggatcctc gaatatgaag atgaagatga aatatttggt gtgtcaaata aaaagcttgt
    13501 gtgcttaagt ttgtgttttt ttcttggctt gttgtgttat gaatttgtgg ctttttctaa
    13561 tatcaaatga atgtaagatc tcattataat gaataaacaa atgtttctat aatccattgt
    13621 gaatgttttg ttggatctct tctgcagcat ataactactg tatgtgctat ggtatggact
    13681 atggaatatg attaaagata aggtttaaac tatcagtgtt tgacaggata tattggcggg
    13741 taaacctaag agaaaagagc gtttattaga ataatcggat atttaaaagg gcgtgaaaag
    13801 gtttatccgt tcgtccattt gtatgtgcat gccaaccaca ggagatccaa aaggatctag
    13861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    13921 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
