LOCUS       sequence-430575-       16441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Binary vector expressing the Lineage Tracing system. Cas9 expression
            driven by CLV3 promoter. LT0 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16441)
  AUTHORS   Dona M, Bradamante G, Bogojevic Z, Gutzat R, Streubel S, Mosiolek M,
            Dolan L, Mittelsten Scheid O
  TITLE     A versatile CRISPR-based system for lineage tracing in living 
            plants.
  JOURNAL   Plant J. 2023 Jul 5. doi: 10.1111/tpj.16378.
  PUBMED    37403571
REFERENCE   2  (bases 1 to 16441)
  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 J. 
            2023 Jul 5. doi: 10.1111/tpj.16378."
COMMENT     Sequence Label: pCLV3Cas-LT0
FEATURES             Location/Qualifiers
     source          1..16441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..86)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(240..828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(320..339)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1042..1066
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     1119..1141
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2277..6380
                     /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             2277..6380
                     /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             6393..6413
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        7402..7477
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(9040..9042,9043..9045,9046..9756)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(9085..9106)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     9693..9714
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             10941..11651
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(11086..11104)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11354..11373
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    11895..11942
                     /label=FRT (mutant)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT site with a mutation in the core sequence (Azam 
                     et al., 1997)"
     misc_feature    12054..12078
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      12158..12789
                     /label=oriV
                     /note="incP origin of replication"
     CDS             14200..14994
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             15293..16441
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MAQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 ggccttgtgg ggtcagttcc ggctgggggt tcagcagcca gcgctttact gagatcctct
       61 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      661 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      841 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      901 atgagattat caaaaaggat cttcacctag atccttttgg atctcctgtg gttggcatgc
      961 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta
     1021 ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt
     1081 aaactgaagg cgggaaacga caatctgcta gtggatctcc cagtcacgac gttgtaaaac
     1141 gggcgccccg cggaaagctt gctagccaat tggggcccaa cgttctcgag tttttctaga
     1201 aggccttgga tccacccggg agaattcgtc gactttgcgg ccgcatcgat actgcaggag
     1261 ctcggtacct tttactagtg atatcgatca tgagcgacct aaaaattacg gatatgaata
     1321 taggcatatc cgtatccgaa ttatccgttt gacagctagc aacgattgta caattgcttc
     1381 tttaaaaaag gaagaaagaa agaaagaaaa gaatcaacat cagcgttaac aaacggcccc
     1441 gttacggccc aaacggtcat atagagtaac ggcgttaagc gttgaaagac tcctatcgaa
     1501 atacgtaacc gcaaacgtgt catagtcaga tcccctcttc cttcaccgcc tcaaacacaa
     1561 aaataatctt ctacagccta tatatacaac ccccccttct atctctcctt tctcacaatt
     1621 catcatcttt ctttctctac ccccaatttt aagaaatcct ctcttctcct cttcattttc
     1681 aaggtaaatc tctctctctc tctctctctc tgttattcct tgttttaatt aggtatgtat
     1741 tattgctagt ttgttaatct gcttatctta tgtatgcctt atgtgaatat ctttatcttg
     1801 ttcatctcat ccgtttagaa gctataaatt tgttgatttg actgtgtatc tacacgtggt
     1861 tatgtttata tctaatcaga tatgaatttc ttcatattgt tgcgtttgtg tgtaccaatc
     1921 cgaaatcgtt gatttttttc atttaatcgt gtagctaatt gtacgtatac atatggatct
     1981 acgtatcaat tgttcatctg tttgtgtttg tatgtataca gatctgaaaa catcacttct
     2041 ctcatctgat tgtgttgtta catacataga tatagatctg ttatatcatt ttttttatta
     2101 attgtgtata tatatatgtg catagatctg gattacatga ttgtgattat ttacatgatt
     2161 ttgttattta cgtatgtata tatgtagatc tggacttttt ggagttgttg acttgattgt
     2221 atttgtgtgt gtatatgtgt gttctgatct tgatatgtta tgtatgtgca gcaacaatgg
     2281 ataagaagta ctctatcgga ctcgatatcg gaactaactc tgtgggatgg gctgtgatca
     2341 ccgatgagta caaggtgcca tctaagaagt tcaaggttct cggaaacacc gataggcact
     2401 ctatcaagaa aaaccttatc ggtgctctcc tcttcgattc tggtgaaact gctgaggcta
     2461 ccagactcaa gagaaccgct agaagaaggt acaccagaag aaagaacagg atctgctacc
     2521 tccaagagat cttctctaac gagatggcta aagtggatga ttcattcttc cacaggctcg
     2581 aagagtcatt cctcgtggaa gaagataaga agcacgagag gcaccctatc ttcggaaaca
     2641 tcgttgatga ggtggcatac cacgagaagt accctactat ctaccacctc agaaagaagc
     2701 tcgttgattc tactgataag gctgatctca ggctcatcta cctcgctctc gctcacatga
     2761 tcaagttcag aggacacttc ctcatcgagg gtgatctcaa ccctgataac tctgatgtgg
     2821 ataagttgtt catccagctc gtgcagacct acaaccagct tttcgaagag aaccctatca
     2881 acgcttcagg tgtggatgct aaggctatcc tctctgctag gctctctaag tcaagaaggc
     2941 ttgagaacct cattgctcag ctccctggtg agaagaagaa cggacttttc ggaaacttga
     3001 tcgctctctc tctcggactc acccctaact tcaagtctaa cttcgatctc gctgaggatg
     3061 caaagctcca gctctcaaag gatacctacg atgatgatct cgataacctc ctcgctcaga
     3121 tcggagatca gtacgctgat ttgttcctcg ctgctaagaa cctctctgat gctatcctcc
     3181 tcagtgatat cctcagagtg aacaccgaga tcaccaaggc tccactctca gcttctatga
     3241 tcaagagata cgatgagcac caccaggatc tcacacttct caaggctctt gttagacagc
     3301 agctcccaga gaagtacaaa gagattttct tcgatcagtc taagaacgga tacgctggtt
     3361 acatcgatgg tggtgcatct caagaagagt tctacaagtt catcaagcct atcctcgaga
     3421 agatggatgg aaccgaggaa ctcctcgtga agctcaatag agaggatctt ctcagaaagc
     3481 agaggacctt cgataacgga tctatccctc atcagatcca cctcggagag ttgcacgcta
     3541 tccttagaag gcaagaggat ttctacccat tcctcaagga taacagggaa aagattgaga
     3601 agattctcac cttcagaatc ccttactacg tgggacctct cgctagagga aactcaagat
     3661 tcgcttggat gaccagaaag tctgaggaaa ccatcacccc ttggaacttc gaagaggtgg
     3721 tggataaggg tgctagtgct cagtctttca tcgagaggat gaccaacttc gataagaacc
     3781 ttccaaacga gaaggtgctc cctaagcact ctttgctcta cgagtacttc accgtgtaca
     3841 acgagttgac caaggttaag tacgtgaccg agggaatgag gaagcctgct tttttgtcag
     3901 gtgagcaaaa gaaggctatc gttgatctct tgttcaagac caacagaaag gtgaccgtga
     3961 agcagctcaa agaggattac ttcaagaaaa tcgagtgctt cgattcagtt gagatttctg
     4021 gtgttgagga taggttcaac gcatctctcg gaacctacca cgatctcctc aagatcatta
     4081 aggataagga tttcttggat aacgaggaaa acgaggatat cttggaggat atcgttctta
     4141 ccctcaccct ctttgaagat agagagatga ttgaagaaag gctcaagacc tacgctcatc
     4201 tcttcgatga taaggtgatg aagcagttga agagaagaag atacactggt tggggaaggc
     4261 tctcaagaaa gctcattaac ggaatcaggg ataagcagtc tggaaagaca atccttgatt
     4321 tcctcaagtc tgatggattc gctaacagaa acttcatgca gctcatccac gatgattctc
     4381 tcacctttaa agaggatatc cagaaggctc aggtttcagg acagggtgat agtctccatg
     4441 agcatatcgc taacctcgct ggatctcctg caatcaagaa gggaatcctc cagactgtga
     4501 aggttgtgga tgagttggtg aaggtgatgg gaaggcataa gcctgagaac atcgtgatcg
     4561 aaatggctag agagaaccag accactcaga agggacagaa gaactctagg gaaaggatga
     4621 agaggatcga ggaaggtatc aaagagcttg gatctcagat cctcaaagag caccctgttg
     4681 agaacactca gctccagaat gagaagctct acctctacta cctccagaac ggaagggata
     4741 tgtatgtgga tcaagagttg gatatcaaca ggctctctga ttacgatgtt gatcatatcg
     4801 tgccacagtc attcttgaag gatgattcta tcgataacaa ggtgctcacc aggtctgata
     4861 agaacagggg taagagtgat aacgtgccaa gtgaagaggt tgtgaagaaa atgaagaact
     4921 attggaggca gctcctcaac gctaagctca tcactcagag aaagttcgat aacttgacta
     4981 aggctgagag gggaggactc tctgaattgg ataaggcagg attcatcaag aggcagcttg
     5041 tggaaaccag gcagatcact aagcacgttg cacagatcct cgattctagg atgaacacca
     5101 agtacgatga gaacgataag ttgatcaggg aagtgaaggt tatcaccctc aagtcaaagc
     5161 tcgtgtctga tttcagaaag gatttccaat tctacaaggt gagggaaatc aacaactacc
     5221 accacgctca cgatgcttac cttaacgctg ttgttggaac cgctctcatc aagaagtatc
     5281 ctaagctcga gtcagagttc gtgtacggtg attacaaggt gtacgatgtg aggaagatga
     5341 tcgctaagtc tgagcaagag atcggaaagg ctaccgctaa gtatttcttc tactctaaca
     5401 tcatgaattt cttcaagacc gagattaccc tcgctaacgg tgagatcaga aagaggccac
     5461 tcatcgagac aaacggtgaa acaggtgaga tcgtgtggga taagggaagg gatttcgcta
     5521 ccgttagaaa ggtgctctct atgccacagg tgaacatcgt taagaaaacc gaggtgcaga
     5581 ccggtggatt ctctaaagag tctatcctcc ctaagaggaa ctctgataag ctcattgcta
     5641 ggaagaagga ttgggaccct aagaaatacg gtggtttcga ttctcctacc gtggcttact
     5701 ctgttctcgt tgtggctaag gttgagaagg gaaagagtaa gaagctcaag tctgttaagg
     5761 aacttctcgg aatcactatc atggaaaggt catctttcga gaagaaccca atcgatttcc
     5821 tcgaggctaa gggatacaaa gaggttaaga aggatctcat catcaagctc ccaaagtact
     5881 cactcttcga actcgagaac ggtagaaaga ggatgctcgc ttctgctggt gagcttcaaa
     5941 agggaaacga gcttgctctc ccatctaagt acgttaactt tctttacctc gcttctcact
     6001 acgagaagtt gaagggatct ccagaagata acgagcagaa gcaacttttc gttgagcagc
     6061 acaagcacta cttggatgag atcatcgagc agatctctga gttctctaaa agggtgatcc
     6121 tcgctgatgc aaacctcgat aaggtgttgt ctgcttacaa caagcacaga gataagccta
     6181 tcagggaaca ggcagagaac atcatccatc tcttcaccct taccaacctc ggtgctcctg
     6241 ctgctttcaa gtacttcgat acaaccatcg ataggaagag atacacctct accaaagaag
     6301 tgctcgatgc taccctcatc catcagtcta tcactggact ctacgagact aggatcgatc
     6361 tctcacagct cggtggtgat tcaagggctg atcctaagaa gaagaggaag gtttgagggg
     6421 ctcacgacga actctaagag ctcaggcctc ccagctttcg tccgtatcat cggtttcgac
     6481 aacgttcgtc aagttcaatg catcagtttc attgcccaca caccagaatc ctactaagtt
     6541 tgagtattat ggcattggaa aagctgtttt cttctatcat ttgttctgct tgtaatttac
     6601 tgtgttcttt cagtttttgt tttcggacat caaaatgcaa atggatggat aagagttaat
     6661 aaatgatatg gtccttttgt tcattctcaa attattatta tctgttgttt ttactttaat
     6721 gggttgaatt taagtaagaa aggaactaac agtgtgatat taaggtgcaa tgttagacat
     6781 ataaaacagt ctttcacctc tctttggtta tgtcttgaat tggtttgttt cttcacttat
     6841 ctgtgtaatc aagtttacta tgagtctatg atcaagtaat tatgcaatca agttaagtac
     6901 agtataggct ttcagctttt tttcttcttc ttcgttcata cagttttttt ttgtttatca
     6961 gcttacattt tcttgaaccg tagctttcgt tttcttcttt ttaactttcc attcggagtt
     7021 tttgtatctt gtttcatagt ttgtcccagg attagaatga ttaggcatcg aaccttcaag
     7081 aatttgattg aataaaacat cttcattctt aagatatgaa gataatcttc aaaaggcccc
     7141 tgggaatctg aaagaagaga agcaggccca tttatatggg aaagaacaat agtatttctt
     7201 atataggccc atttaagttg aaaacaatct tcaaaagtcc cacatcgctt agataagaaa
     7261 acgaagctga gtttatatac agctagagtc gaagtagtga ttggaacaaa gcaccagtgg
     7321 tctagtggta gaatagtacc ctgccacggt acagacccgg gttcgattcc cggctggtgc
     7381 agtcccctcc accccacagt ggttttagag ctagaaatag caagttaaaa taaggctagt
     7441 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttttttttt ctgctttctg
     7501 cccttgccaa tccatatctg aagaagaaaa aaaaacacaa aattacgatt tagatccgat
     7561 ataacaaaat ttgaatcgca cagatcgatc tctttggaga ttctatacct agaaaatgga
     7621 gacgattttc aaatctctgt aaaaattctg gtttcttctt gacggaagaa gacgacgact
     7681 ccaatatttc ggttagtact gaaccggaaa gtttgactgg tgcaaccaat ttaatgtacc
     7741 gtacgtaacg caccaatcgg attttgtatt caatgggcct tatctgtgag cccattaatt
     7801 gatgtgacgg cctaaactaa atccgaacgg tttatttcag cgatccgcga cggtttgtat
     7861 tcagccaata gcaatcaatt atgtagcagt ggtgatcctc gtcaaaccag taaagctaga
     7921 tctggaccgt tgaattggtg caagaaagca catgttgtga tatttttacc cgtacgatta
     7981 gaaaacttga gaaacacatt gataatcgat aaaaaccgtc cgatcatata aatccgcttt
     8041 accatcgttg cctataaatt aatatcaata gccgtacacg cgtgaagact gacaatatta
     8101 tctttttcga attcggagct caagtttgaa attcggagaa gctagagagt tttctgaggt
     8161 acgattcttc gatcctcttt gattttcctg gaaatatttt ttcggtgatc gtgaaactac
     8221 tggaatcgct cgataggtgg tacgaaatta ggcgagatta gtttctattc ttggccatta
     8281 tcttgtttct tcgccgaatg atcttccgga taaagatttt aggttagaga tgaatcgtat
     8341 agctagattt catcaccaga tagtttcttt gtctagaatc tctgaaattc tcgatagttt
     8401 tcacatgtgt aaatagattg ttcttattcg gcgattgttg attagggttt tgattttctt
     8461 gattatgcga ttgcaattag ggattttctt tggttttgtg ttgatcttac gatacattcc
     8521 ggcaattgaa tacgtatgga tctaaatctt gttaatttgt tgaacaggac gatgcgtccc
     8581 ctccacccca cagtgggggc accaagagcc gagaagaagc ccgccgagaa aaaaaccgcc
     8641 gctgagaggc cggtggaaga gaacaaggct gctgaaaagg ctccggcgga gaagaaaccc
     8701 aaggcgggga agaaattgcc accaaaggaa gccggtgaca agaagaagaa gcgatctaag
     8761 aaaaacgtcg aaacctacaa gatctacatc ttcaaggtgc tgaagcaggt tcatccagat
     8821 atcggaattt caagcaaagc catggggatt atgaacagtt tcatcaacga catattcgag
     8881 aagctagctc aggaatcttc gaaactcgct aggtacaaca agaagccgac cattacatct
     8941 cgcgagatcc agacggctgt gagactcgtc ctccccggtg aactcgctaa acacgccgtc
     9001 tctgaaggta ccaaggcggt taccaagttt actagctcca tggtgagcaa gggcgaggag
     9061 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     9121 ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     9181 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     9241 ggcgtggcct gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     9301 gccatgcccg aaggctacgt ccaggagcgc accatctctt tcaaggacga cggtacctac
     9361 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     9421 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     9481 agccacaacg tctatatcac ggccgacaag cagaagaacg gcatcaaggc taacttcaag
     9541 atccgccaca acgttgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     9601 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcca tcagtccgcc
     9661 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     9721 gccgggatta cacatggcat ggacgagctg tacaagtaat caggtggatc ctagataacc
     9781 tttaccttca ttccctgtaa attggtaccc tgcatctcgt ctctgttatg cttaagaagt
     9841 tcaatgtttc gtttcatgta aaactttggt ggtttgtgtt ttggggcctt gtataatccc
     9901 tgatgaataa gtgttctact atgtttccgt tcctgttatc tctttctttc taatgacaag
     9961 tcgaacttct tctttatcat cgcttcgttt ttattatctg tgcttctttt gtttaatacg
    10021 cctgcaaagt gactcgactc tgtttagtgc agttctgcga aacttgtaaa tagtccaatt
    10081 gttggcctct agtaatagat gtagcgaaag tgttgagctg ttgggttcta aggatggctt
    10141 gaacatgtta atcttttagg ttctgagtat gatgaacatt cgttgttgct aagaaatgcc
    10201 tgtaatgtcc cacaaatgta gaaaatggtt cgtacctttg tccaagcatt gatatgtctg
    10261 atgagaggaa actgcaagat actgagcttg gtttaacgaa ggagaggcag tttcttcctt
    10321 ccaaagcatt tcatttgaca atgccttgat catcttaagt agagtttctg ttgtggaaag
    10381 tttgaaactt tgaagaaacg actctcaagt aaattgatga tcacaagtga aagtgtatgt
    10441 tactaactat tgaaaccaaa ttaacatagg gtttttattt aaataaaagt ttaaccttct
    10501 ttttaaaaaa ttgttcatag tgtcatgtca gaacaagagc tacaaatcac acatagcatg
    10561 cataagcgga gctatgatga gtggtattgt tttgttcgtc acttgtcact cttttccaac
    10621 acataatccc gacaacaacg taagagcatc tctctctctc cacacacact catgcatgca
    10681 tgcattctta cacgtgattg ccatgcaaat ctcctttctc acctataaat acaaaccaac
    10741 ccttcactac actcttcact caaaccaaaa caagaaaaca tacacaaata gcaaaacaag
    10801 ggcgaattct caacacaaca tatacaaaac aaacgaatct caagcaatca agcattctac
    10861 ttctattgca gcaatttaaa tcatttcttt taaagcaaaa gcaattttct gaaaattttc
    10921 accatttacg aacgatagcc atggtgagca agggcgagga ggataacatg gccatcatca
    10981 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
    11041 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
    11101 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
    11161 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
    11221 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
    11281 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
    11341 tcccctccga cggccccgta atgcagaaga agacgatggg ctgggaggcc tcctccgagc
    11401 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
    11461 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
    11521 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
    11581 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
    11641 tgtacaagta gcttggctgc aggtcgacct gcaggcatgc ccgctgaaat caccagtctc
    11701 tctctacaaa tctatctctc tctataataa tgtgtgagta gttcccagat aagggaatta
    11761 gggttcttat agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg
    11821 tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa tccaggggta
    11881 ccgtatgatc tagggaagtt cctattccga agttcctatt ctctgaaaag tataggaact
    11941 tctttgcgta ttgggcgctc ttggcctttt tggccaccgg tcgtacggtt aaaaccaccc
    12001 cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca
    12061 ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa
    12121 tcaccactcg atacaggcag cccatcagtc cactagagac gctcaccggg ctggttgccc
    12181 tcgccgctgg gctggcggcc gtctatggcc ctgcaaacgc gccagaaacg ccgtcgaagc
    12241 cgtgtgcgag acaccgcagc cgccggcgtt gtggatacct cgcggaaaac ttggccctca
    12301 ctgacagatg aggggcggac gttgacactt gaggggccga ctcacccggc gcggcgttga
    12361 cagatgaggg gcaggctcga tttcggccgg cgacgtggag ctggccagcc tcgcaaatcg
    12421 gcgaaaacgc ctgattttac gcgagtttcc cacagatgat gtggacaagc ctggggataa
    12481 gtgccctgcg gtattgacac ttgaggggcg cgactactga cagatgaggg gcgcgatcct
    12541 tgacacttga ggggcagagt gctgacagat gaggggcgca cctattgaca tttgaggggc
    12601 tgtccacagg cagaaaatcc agcatttgca agggtttccg cccgtttttc ggccaccgct
    12661 aacctgtctt ttaacctgct tttaaaccaa tatttataaa ccttgttttt aaccagggct
    12721 gcgccctgtg cgcgtgaccg cgcacgccga aggggggtgc ccccccttct cgaaccctcc
    12781 cggcccgctc tcgcgttggc agcatcaccc ataattgtgg tttcaaaatc ggctccgtcg
    12841 atactatgtt atacgccaac tttgaaaaca actttgaaaa agctgttttc tggtatttaa
    12901 agttttagaa tgcaaggaac agtgaattgg agttcgtctt gttataatta gggaaggtgc
    12961 gaataagcgg ggaaattctt ctcggctgac tcagtcattt catttcttca tgtttgagcc
    13021 gattttttct cccgtaaatg ccttgaatca gcctatttag accgtttctt cgccatttaa
    13081 ggcgttatcc ccagttttta gtgagatctc tcccactgac gtatcatttg gtccgcccga
    13141 aacaggttgg ccagcgtgaa taacatcgcc agttggttat cgtttttcag caaccccttg
    13201 tatctggctt tcacgaagcc gaactgtcgc ttgatgatgc gaaatgggtg ctccaccctg
    13261 gcccggatgc tggctttcat gtattcgatg ttgatggccg ttttgttctt gcgtggatgc
    13321 tgtttcaagg ttcttacctt gccggggcgc tcggcgatca gccagtccac atccacctcg
    13381 gccagctcct cgcgctgtgg cgccccttgg tagccggcat cggctgagac aaattgctcc
    13441 tctccatgca gcagattacc cagctgattg aggtcatgct cgttggccgc ggtggtgacc
    13501 aggctgtggg tcaggccact cttggcatcg acaccaatgt gggccttcat gccaaagtgc
    13561 cactgattgc ctttcttggt ctgatgcatc tccggatcgc gttgctgctc tttgttcttg
    13621 gtcgagctgg gtgcctcaat gatggtggca tcgaccaagg tgccttgagt catcatgacg
    13681 cctgcttcgg ccagccagcg attgatggtc ttgaacaatt ggcgggccag ttgatgctgc
    13741 tccagcaggt ggcggaaatt catgatggtg gtgcggtccg gcaaggcgct atccagggat
    13801 aaccgggcaa acagacgcat ggaggcgatt tcgtacagag catcttccat cgcgccatcg
    13861 ctcaggttgt accaatgctg catgcagtga atgcgtagca tggtttccag cggataaggt
    13921 cgccggccat taccagcctt ggggtaaaac ggctcgatga cttccaccat gttttgccat
    13981 ggcagaatct gctccatgcg ggacaagaaa atctcttttc tggtctgacg gcgcttactg
    14041 ctgaattcac tgtcggcgaa ggtaagttga tgactcatga tgaaccctgt tctatggctc
    14101 cagatgacaa acatgatctc atatcaggga cttgttcgca ccttccttag cttcttgggg
    14161 tatctttaaa tactgtagaa aagaggaagg aaataataaa tggctaaaat gagaatatca
    14221 ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct
    14281 cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac
    14341 agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg
    14401 gaaggaaagc tgcctgttcc aaaggtcctg caccttgaac ggcatgatgg ctggagcaat
    14461 ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc
    14521 cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata
    14581 tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg
    14641 aataacgatc tggccgatgt ggattgcgaa aactgggaag aagacacccc atttaaagat
    14701 ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc
    14761 cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt
    14821 gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg
    14881 atcagggagg atattgggga agaacagtat gtcgagctat tttttgactt actggggatc
    14941 aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt ttagtaccta
    15001 gatgtggcgc aacgatgctg gcgacaagca ggagcgcacc gacttcttcc gcatcaagtg
    15061 ttttggctct caggccgagg cccacggcaa gtatttgggc aaggggtcgc tggtattcgt
    15121 gcagggcaag attcggaata ccaagtacga gaaggacggc cagacggtct acgggaccga
    15181 cttcattgcc gataaggtgg attatctgga caccaaggca ccaggcggat caaatcagga
    15241 ataagggcac attgccccgg cgtgagtcgg ggcaatcccg caaggagggt gaatgaatcg
    15301 gacgtttgac cggaaggcat acaggcaaga actgatcgac gcggggtttt ccgccgagga
    15361 tgccgaaacc atcgcaagcc gcaccgtcat gcgtgcgccc cgcgaaacct tccagtccgt
    15421 cggctcgatg gcccagcaag ctacggccaa gatcgagcgc gacagcgtgc aactggctcc
    15481 ccctgccctg cccgcgccat cggccgccgt ggagcgttcg cgtcgtctcg aacaggaggc
    15541 ggcaggtttg gcgaagtcga tgaccatcga cacgcgagga actatgacga ccaagaagcg
    15601 aaaaaccgcc ggcgaggacc tggcaaaaca ggtcagcgag gccaagcaag ccgcgttgct
    15661 gaaacacacg aagcagcaga tcaaggaaat gcagctttcc ttgttcgata ttgcgccgtg
    15721 gccggacacg atgcgagcga tgccaaacga cacggcccgc tctgccctgt tcaccacgcg
    15781 caacaagaaa atcccgcgcg aggcgctgca aaacaaggtc attttccacg tcaacaagga
    15841 cgtgaagatc acctacaccg gcgtcgagct gcgggccgac gatgacgaac tggtgtggca
    15901 gcaggtgttg gagtacgcga agcgcacccc tatcggcgag ccgatcacct tcacgttcta
    15961 cgagctttgc caggacctgg gctggtcgat caatggccgg tattacacga aggccgagga
    16021 atgcctgtcg cgcctacagg cgacggcgat gggcttcacg tccgaccgcg ttgggcacct
    16081 ggaatcggtg tcgctgctgc accgcttccg cgtcctggac cgtggcaaga aaacgtcccg
    16141 ttgccaggtc ctgatcgacg aggaaatcgt cgtgctgttt gctggcgacc actacacgaa
    16201 attcatatgg gagaagtacc gcaagctgtc gccgacggcc cgacggatgt tcgactattt
    16261 cagctcgcac cgggagccgt acccgctcaa gctggaaacc ttccgcctca tgtgcggatc
    16321 ggattccacc cgcgtgaaga agtggcgcga gcaggtcggc gaagcctgcg aagagttgcg
    16381 aggcagcggc ctggtggaac acgcctgggt caatgatgac ctggtgcatt gcaaacgcta
    16441 g
//
