LOCUS       sequence-203121-       18527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 1xplex gRNA targeting Glyma.04g057400 (Dcl3a-2). Also 
            expresses Cas9 from Gmubi promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18527)
  AUTHORS   Curtin SJ, Xiong Y, Michno JM, Campbell BW, Stec AO, Cermak T, 
            Starker C, Voytas DF, Eamens AL, Stupar RM
  TITLE     CRISPR/Cas9 and TALENs generate heritable mutations for genes 
            involved in small RNA processing of Glycine max and Medicago 
            truncatula.
  JOURNAL   Plant Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857.
  PUBMED    29087011
REFERENCE   2  (bases 1 to 18527)
  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 
            Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857."
COMMENT     Sequence Label: pSC45
FEATURES             Location/Qualifiers
     source          1..18527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(127..498)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            531..640
                     /note="incP origin of transfer"
     primer_bind     863..882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1116..1133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1321..1971)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2436..2460
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3668..3692
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3714..3761
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3768..3788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3813..5249,5439..8102)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system"
                     /label=pcoCas9
                     /note="plant codon-optimized Cas9 gene containing the 
                     potato IV2 intron"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     intron          5250..5438
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8103..8150
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8161..8413
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8458..8478
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8960..9035
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9047..9071)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9090..9114)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9152..9859
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10409..10754
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     10697..10717
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             10775..11326
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      11470..11722
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(11735..11752)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11735..11751)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11744..11766)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(11891..11910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12101..12122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12369..12393
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(13989..15137)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(15436..16230)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16422..17189
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      17882..18517
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg
       61 atggagcgca tggggacgtg cttggcaatc acgcgcaccc cccggccgtt ttagcggcta
      121 aaaaagtcat ggctctgccc tcgggcggac cacgcccatc atgaccttgc caagctcgtc
      181 ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga accgcgccgt
      241 gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca ggtcgccatt
      301 gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt tgtagccctg
      361 gccgacggcc agcaggtagg ccgacaggct catgccggcc gccgccgcct tttcctcaat
      421 cgctcttcgt tcgtctggaa ggcagtacac cttgataggt gggctgccct tcctggttgg
      481 cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag ccggccagcc
      541 tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag tgaagaagga
      601 acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc gttggataca
      661 ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga aaaaggatgg
      721 atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg aaaagcgcca
      781 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      841 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      901 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      961 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1021 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     1081 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1141 aagagcgcca gaaggccgcc agagaggccg agcgcggccg tgaggcttgg acgctagggc
     1201 agggcatgaa aaagcccgta gcgggctgct acgggcgtct gacgcggtgg aaagggggag
     1261 gggatgttgt ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga
     1321 tcaatcgtca ccctttctcg gtccttcaac gttcctgaca acgagcctcc ttttcgccaa
     1381 tccatcgaca atcaccgcga gtccctgctc gaacgctgcg tccggaccgg cttcgtcgaa
     1441 ggcgtctatc gcggcccgca acagcggcga gagcggagcc tgttcaacgg tgccgccgcg
     1501 ctcgccggca tcgctgtcgc cggcctgctc ctcaagcacg gccccaacag tgaagtagct
     1561 gattgtcatc agcgcattga cggcgtcccc ggccgaaaaa cccgcctcgc agaggaagcg
     1621 aagctgcgcg tcggccgttt ccatctgcgg tgcgcccggt cgcgtgccgg catggatgcg
     1681 cgcgccatcg cggtaggcga gcagcgcctg cctgaagctg cgggcattcc cgatcagaaa
     1741 tgagcgccag tcgtcgtcgg ctctcggcac cgaatgcgta tgattctccg ccagcatggc
     1801 ttcggccagt gcgtcgagca gcgcccgctt gttcctgaag tgccagtaaa gcgccggctg
     1861 ctgaaccccc aaccgttccg ccagtttgcg tgtcgtcaga ccgtctacgc cgacctcgtt
     1921 caacaggtcc agggcggcac ggatcactgt attcggctgc aactttgtca tgcttgacac
     1981 tttatcactg ataaacataa tatgtccacc aacttatcag tgataaagaa tccgcgcgtt
     2041 caatcggacc agcggaggct ggtccggagg ccagacgtga aacccaacat acccctgatc
     2101 gtaattctga gcactgtcgc gctcgacgct gtcggcatcg gcctgattat gccggtgctg
     2161 ccgggcctcc tgcgcgatct ggttcactcg aacgacgtca ccgcccacta tggcattctg
     2221 ctggcgctgt atgcgttggt gcaatttgcc tgcgcacctg tgctgggcgc gctgtcggat
     2281 cgtttcgggc ggcggccaat cttgctcgtc tcgctggccg gcgccagatc tggggaaccc
     2341 tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat
     2401 atccgattat tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca
     2461 aacactgata gtttaaactg aaggcgggaa acgacaatct gatcatgagc ggagaattaa
     2521 gggagtcacg ttatgacccc cgccgatgac gcgggacaag ccgttttacg tttggaactg
     2581 acagaaccgc aacgttgaag gagccactca gccgcgggtt tctggagttt aatgagctaa
     2641 gcacatacgt cagaaaccat tattgcgcgt tcaaaagtcg cctaaggtca ctatcagcta
     2701 gacggacgcg gccgcgggcc caatataaca acgacgtcgt aacagataaa gcgaagcttg
     2761 aaggtgcatg tgactccgtc aagattacga aaccgccaac taccacgcaa attgcaattc
     2821 tcaatttcct agaaggactc tccgaaaatg catccaatac caaatattac ccgtgtcata
     2881 ggcaccaagt gacaccatac atgaacacgc gtcacaatat gactggagaa gggttccaca
     2941 ccttatgcta taaaacgccc cacacccctc ctccttcctt cgcagttcaa ttccaatata
     3001 ttccattctc tctgtgtatt tccctacctc tcccttcaag gttagtcgat ttcttctgtt
     3061 tttcttcttc gttctttcca tgaattgtgt atgttctttg atcaatacga tgttgatttg
     3121 attgtgtttt gtttggtttc atcgatcttc aattttcata atcagattca gcttttatta
     3181 tctttacaac aacgtcctta atttgatgat tctttaatcg tagatttgct ctaattagag
     3241 ctttttcatg tcagatccct ttacaacaag ccttaattgt tgattcatta atcgtagatt
     3301 agggcttttt tcattgatta cttcagatcc gttaaacgta accatagatc agggcttttt
     3361 catgaattac ttcagatccg ttaaacaaca gccttatttt ttatacttct gtggtttttc
     3421 aagaaattgt tcagatccgt tgacaaaaag ccttattcgt tgattctata tcgtttttcg
     3481 agagatattg ctcagatctg ttagcaactg ccttgtttgt tgattctatt gccgtggatt
     3541 agggtttttt ttcacgagat tgcttcagat ccgtacttaa gattacgtaa tggattttga
     3601 ttctgattta tctgtgattg ttgactcgac aggtaccttc aaacctcgag gcgcgccaag
     3661 ctatcaaaca agtttgtaca aaaaagcagg ctccgaattc gcccttcacc atggattaca
     3721 aggatgatga tgataaggat tacaaggatg atgatgataa gatggctcca aagaagaaga
     3781 gaaaggttgg aatccacgga gttccagctg ctgataagaa gtactctatc ggacttgaca
     3841 tcggaaccaa ctctgttgga tgggctgtta tcaccgatga gtacaaggtt ccatctaaga
     3901 agttcaaggt tcttggaaac accgatagac actctatcaa gaagaacctt atcggtgctc
     3961 ttcttttcga ttctggagag accgctgagg ctaccagatt gaagagaacc gctagaagaa
     4021 gatacaccag aagaaagaac agaatctgct accttcagga aatcttctct aacgagatgg
     4081 ctaaggttga tgattctttc ttccacagac ttgaggagtc tttccttgtt gaggaggata
     4141 agaagcacga gagacaccca atcttcggaa acatcgttga tgaggttgct taccacgaga
     4201 agtacccaac catctaccac cttagaaaga agttggttga ttctaccgat aaggctgatc
     4261 ttagacttat ctaccttgct cttgctcaca tgatcaagtt cagaggacac ttccttatcg
     4321 agggagacct taacccagat aactctgatg ttgataagtt gttcatccag cttgttcaga
     4381 cctacaacca gcttttcgag gagaacccaa tcaacgcttc tggagttgat gctaaggcta
     4441 tcctttctgc tagactttct aagtctcgta gacttgagaa ccttatcgct cagcttccag
     4501 gagagaagaa gaacggactt ttcggaaacc ttatcgctct ttctcttgga cttaccccaa
     4561 acttcaagtc taacttcgat cttgctgagg atgctaagtt gcagctttct aaggatacct
     4621 acgatgatga tcttgataac cttcttgctc agatcggaga tcagtacgct gatcttttcc
     4681 ttgctgctaa gaacctttct gatgctatcc ttctttctga catccttaga gttaacaccg
     4741 agatcaccaa ggctccactt tctgcttcta tgatcaagag atacgatgag caccaccagg
     4801 atcttaccct tttgaaggct cttgttagac agcagcttcc agagaagtac aaggaaatct
     4861 tcttcgatca gtctaagaac ggatacgctg gatacatcga tggaggagct tctcaggagg
     4921 agttctacaa gttcatcaag ccaatccttg agaagatgga tggaaccgag gagcttcttg
     4981 ttaagttgaa cagagaggat cttcttagaa agcagagaac cttcgataac ggatctatcc
     5041 cacaccagat ccaccttgga gagcttcacg ctatccttcg tagacaggag gatttctacc
     5101 cattcttgaa ggataacaga gagaagatcg agaagatcct taccttcaga atcccatact
     5161 acgttggacc acttgctaga ggaaactctc gtttcgcttg gatgaccaga aagtctgagg
     5221 agaccatcac cccttggaac ttcgaggagg taagtttctg cttctacctt tgatatatat
     5281 ataataatta tcattaatta gtagtaatat aatatttcaa atattttttt caaaataaaa
     5341 gaatgtagta tatagcaatt gcttttctgt agtttataag tgtgtatatt ttaatttata
     5401 acttttctaa tatatgacca aaatttgttg atgtgcaggt tgttgataag ggagcttctg
     5461 ctcagtcttt catcgagaga atgaccaact tcgataagaa ccttccaaac gagaaggttc
     5521 ttccaaagca ctctcttctt tacgagtact tcaccgttta caacgagctt accaaggtta
     5581 agtacgttac cgagggaatg agaaagccag ctttcctttc tggagagcag aagaaggcta
     5641 tcgttgatct tcttttcaag accaacagaa aggttaccgt taagcagttg aaggaggatt
     5701 acttcaagaa gatcgagtgc ttcgattctg ttgaaatctc tggagttgag gatagattca
     5761 acgcttctct tggaacctac cacgatcttt tgaagatcat caaggataag gatttccttg
     5821 ataacgagga gaacgaggac atccttgagg acatcgttct tacccttacc cttttcgagg
     5881 atagagagat gatcgaggag agactcaaga cctacgctca ccttttcgat gataaggtta
     5941 tgaagcagtt gaagagaaga agatacaccg gatggggtag actttctcgt aagttgatca
     6001 acggaatcag agataagcag tctggaaaga ccatccttga tttcttgaag tctgatggat
     6061 tcgctaacag aaacttcatg cagcttatcc acgatgattc tcttaccttc aaggaggaca
     6121 tccagaaggc tcaggtttct ggacagggag attctcttca cgagcacatc gctaaccttg
     6181 ctggatctcc agctatcaag aagggaatcc ttcagaccgt taaggttgtt gatgagcttg
     6241 ttaaggttat gggtagacac aagccagaga acatcgttat cgagatggct agagagaacc
     6301 agaccaccca gaagggacag aagaactctc gtgagagaat gaagagaatc gaggagggaa
     6361 tcaaggagct tggatctcaa atcttgaagg agcacccagt tgagaacacc cagcttcaga
     6421 acgagaagtt gtacctttac taccttcaga acggaagaga tatgtacgtt gatcaggagc
     6481 ttgacatcaa cagactttct gattacgatg ttgatcacat cgttccacag tctttcttga
     6541 aggatgattc tatcgataac aaggttctta cccgttctga taagaacaga ggaaagtctg
     6601 ataacgttcc atctgaggag gttgttaaga agatgaagaa ctactggaga cagcttctta
     6661 acgctaagtt gatcacccag agaaagttcg ataaccttac caaggctgag agaggaggac
     6721 tttctgagct tgataaggct ggattcatca agagacagct tgttgagacc agacagatca
     6781 ccaagcacgt tgctcagatc cttgattctc gtatgaacac caagtacgat gagaacgata
     6841 agttgatcag agaggttaag gttatcacct tgaagtctaa gttggtttct gatttcagaa
     6901 aggatttcca gttctacaag gttagagaga tcaacaacta ccaccacgct cacgatgctt
     6961 accttaacgc tgttgttgga accgctctta tcaagaagta cccaaagttg gagtctgagt
     7021 tcgtttacgg agattacaag gtttacgatg ttagaaagat gatcgctaag tctgagcagg
     7081 agatcggaaa ggctaccgct aagtacttct tctactctaa catcatgaac ttcttcaaga
     7141 ccgagatcac ccttgctaac ggagagatca gaaagagacc acttatcgag accaacggag
     7201 agaccggaga gatcgtttgg gataagggaa gagatttcgc taccgttaga aaggttcttt
     7261 ctatgccaca ggttaacatc gttaagaaaa ccgaggttca gaccggagga ttctctaagg
     7321 agtctatcct tccaaagaga aactctgata agttgatcgc tagaaagaag gattgggacc
     7381 caaagaagta cggaggattc gattctccaa ccgttgctta ctctgttctt gttgttgcta
     7441 aggttgagaa gggaaagtct aagaagttga agtctgttaa ggagcttctt ggaatcacca
     7501 tcatggagcg ttcttctttc gagaagaacc caatcgattt ccttgaggct aagggataca
     7561 aggaggttaa gaaggatctt atcatcaagt tgccaaagta ctctcttttc gagcttgaga
     7621 acggaagaaa gagaatgctt gcttctgctg gagagcttca gaagggaaac gagcttgctc
     7681 ttccatctaa gtacgttaac ttcctttacc ttgcttctca ctacgagaag ttgaagggat
     7741 ctccagagga taacgagcag aagcagcttt tcgttgagca gcacaagcac taccttgatg
     7801 agatcatcga gcaaatctct gagttctcta agagagttat ccttgctgat gctaaccttg
     7861 ataaggttct ttctgcttac aacaagcaca gagataagcc aatcagagag caggctgaga
     7921 acatcatcca ccttttcacc cttaccaacc ttggtgctcc agctgctttc aagtacttcg
     7981 ataccaccat cgatagaaaa agatacacct ctaccaagga ggttcttgat gctaccctta
     8041 tccaccagtc tatcaccgga ctttacgaga ccagaatcga tctttctcag cttggaggag
     8101 ataagagacc agctgctacc aagaaggctg gacaggctaa gaagaagaag tgagacgtcc
     8161 gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg
     8221 atgattatca tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc
     8281 atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac
     8341 gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct
     8401 atgttactag atcgggaatt gatcccccct cgacagcttc cggaaagggc gaattcgcaa
     8461 ctttgtatac aaaagttgcc gagctcgcgg ccgccttcgt tgaacaacgg aaactcgact
     8521 tgccttccgc acaatacatc atttcttctt agcttttttt cttcttcttc gttcatacag
     8581 tttttttttg tttatcagct tacattttct tgaaccgtag ctttcgtttt cttcttttta
     8641 actttccatt cggagttttt gtatcttgtt tcatagtttg tcccaggatt agaatgatta
     8701 ggcatcgaac cttcaagaat ttgattgaat aaaacatctt cattcttaag atatgaagat
     8761 aatcttcaaa aggcccctgg gaatctgaaa gaagagaagc aggcccattt atatgggaaa
     8821 gaacaatagt atttcttata taggcccatt taagttgaaa acaatcttca aaagtcccac
     8881 atcgcttaga taagaaaacg aagctgagtt tatatacagc tagagtcgaa gtagtgattg
     8941 actgtgttga agctctgatg ttttagagct agaaatagca agttaaaata aggctagtcc
     9001 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg
     9061 tacaaagttg gcattactgc aggtcgacga cccagctttc ttgtacaaag tggttcgata
     9121 attcttaatt aactagttct aatctagagt cctgctttaa tgagatatgc gagacgccta
     9181 tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg
     9241 tagctcagat ccttaccgcc ggtttcggtt cattctaatg aatatatcac ccgttactat
     9301 cgtattttta tgaataatat tctccgttca atttactgat tgtaccctac tacttatatg
     9361 tacaatatta aaatgaaaac aatatattgt gctgaatagg tttatagcga catctatgat
     9421 agagcgccac aataacaaac aattgcgttt tattattaca aatccaattt taaaaaaagc
     9481 ggcagaaccg gtcaaaccta aaagactgat tacataaatc ttattcaaat ttcaaaagtg
     9541 ccccaggggc tagtatctac gacacaccga gcggcgaact aataacgctc actgaaggga
     9601 actccggttc cccgccggcg cgcatgggtg agattccttg aagttgagta ttggccgtcc
     9661 gctctaccga aagttacggg caccattcaa cccggtccag cacggcggcc gggtaaccga
     9721 cttgctgccc cgagaattat gcagcatttt tttggtgtat gtgggcccca aatgaagtgc
     9781 aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt ccagggcgaa ttttgcgaca
     9841 acatgtcgag gctcagcagg acctgcaggc atgcaagcta gtgaagaatc gatttgctac
     9901 gtattgcccc tgcaggtccc cagattagcc ttttcaattt cagaaagaat gctaacccac
     9961 agatggttag agaggcttac gcagcaggtc tcatcaagac gatctacccg agcaataatc
    10021 tccaggaaat caaatacctt cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta
    10081 actgcatcaa gaacacagag aaagatatat ttctcaagat cagaagtact attccagtat
    10141 ggacgattca aggcttgctt cacaaaccaa ggcaagtaat agagattgga gtctctaaaa
    10201 aggtagttcc cactgaatca aaggccatgg agtcaaagat tcaaatagag gacctaacag
    10261 aactcgccgt aaagactggc gaacagttca tacagagtct cttacgactc aatgacaaga
    10321 agaaaatctt cgtcaacatg gtggagcacg acacacttgt ctactccaaa aatatcaaag
    10381 atacagtctc agaagaccaa agggcaattg agacttttca acaaagggta atatccggaa
    10441 acctcctcgg attccattgc ccagctatct gtcactttat tgtgaagata gtggaaaagg
    10501 aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt gaagatgcct
    10561 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
    10621 acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg
    10681 atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc
    10741 atttggagag aacacggggg actctagagg atccatgagc ccagaacgac gcccggccga
    10801 catccgccgt gccaccgagg cggacatgcc ggcggtctgc accatcgtca accactacat
    10861 cgagacaagc acggtcaact tccgtaccga gccgcaggaa ccgcaggagt ggacggacga
    10921 cctcgtccgt ctgcgggagc gctatccctg gctcgtcgcc gaggtggacg gcgaggtcgc
    10981 cggcatcgcc tacgcgggcc cctggaaggc acgcaacgcc tacgactgga cggccgagtc
    11041 gaccgtgtac gtctcccccc gccaccagcg gacgggactg ggctccacgc tctacaccca
    11101 cctgctgaag tccctggagg cacagggctt caagagcgtg gtcgctgtca tcgggctgcc
    11161 caacgacccg agcgtgcgca tgcacgaggc gctcggatat gccccccgcg gcatgctgcg
    11221 ggcggccggc ttcaagcacg ggaactggca tgacgtgggt ttctggcagc tggacttcag
    11281 cctgccggta ccgccccgtc cggtcctgcc cgtcaccgag atctgaacgg agtgcgcgtg
    11341 gcatcgcccg agttggagct ggtacgggaa ctcatcgaac tcaactggca tacccgcaat
    11401 ggtgaggtgg aaccgcggcg gatcgcgtac gaccgtgccc aggagggggt accgagctcg
    11461 aatttccccg atcgttcaaa catttggcaa taaagtttct taagattgaa tcctgttgcc
    11521 ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt aataattaac
    11581 atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc gcaattatac
    11641 atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt atcgcgcgcg
    11701 gtgtcatcta tgttactaga tcgggaatta attcactggc cgtcgtttta caacgtcgtg
    11761 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
    11821 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
    11881 atggcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    11941 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    12001 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    12061 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    12121 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
    12181 tcggaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
    12241 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
    12301 aaagaaaaac caccccagta cattaaaaac gtccgcaatg tgttattaag ttgtctaagc
    12361 gtcaatttgt ttacaccaca atatatcctg ccaccagcca gccaacagct ccccgaccgg
    12421 cagctcggca caaaatcacc actcgataca ggcagcccat cagtccggga cggcgtcagc
    12481 gggagagccg ttgtaaggcg gcagactttg ctcatgttac cgatgctatt cggaagaacg
    12541 gcaactaagc tgccgggttt gaaacacgga tgatctcgcg gagggtagca tgttgattgt
    12601 aacgatgaca gagcgttgct gcctgtgatc aaatatcatc tccctcgcag agatccgaat
    12661 tatcagcctt cttattcatt tctcgcttaa ccgtgacagg ctgtcgatct tgagaactat
    12721 gccgacataa taggaaatcg ctggataaag ccgctgagga agctgagtgg cgctatttct
    12781 ttagaagtga acgttgacga tatcaactcc cctatccatt gctcaccgaa tggtacaggt
    12841 cggggacccg aagttccgac tgtcggcctg atgcatcccc ggctgatcga ccccagatct
    12901 ggggctgaga aagcccagta aggaaacaac tgtaggttcg agtcgcgaga tcccccggaa
    12961 ccaaaggaag taggttaaac ccgctccgat caggccgagc cacgccaggc cgagaacatt
    13021 ggttcctgta ggcatcggga ttggcggatc aaacactaaa gctactggaa cgagcagaag
    13081 tcctccggcc gccagttgcc aggcggtaaa ggtgagcaga ggcacgggag gttgccactt
    13141 gcgggtcagc acggttccga acgccatgga aaccgccccc gccaggcccg ctgcgacgcc
    13201 gacaggatct agcgctgcgt ttggtgtcaa caccaacagc gccacgcccg cagttccgca
    13261 aatagccccc aggaccgcca tcaatcgtat cgggctacct agcagagcgg cagagatgaa
    13321 cacgaccatc agcggctgca cagcgcctac cgtcgccgcg accccgcccg gcaggcggta
    13381 gaccgaaata aacaacaagc tccagaatag cgaaatatta agtgcgccga ggatgaagat
    13441 gcgcatccac cagattcccg ttggaatctg tcggacgatc atcacgagca ataaacccgc
    13501 cggcaacgcc cgcagcagca taccggcgac ccctcggcct cgctgttcgg gctccacgaa
    13561 aacgccggac agatgcgcct tgtgagcgtc cttggggccg tcctcctgtt tgaagaccga
    13621 cagcccaatg atctcgccgt cgatgtaggc gccgaatgcc acggcatctc gcaaccgttc
    13681 agcgaacgcc tccatgggct ttttctcctc gtgctcgtaa acggacccga acatctctgg
    13741 agctttcttc agggccgaca atcggatctc gcggaaatcc tgcacgtcgg ccgctccaag
    13801 ccgtcgaatc tgagccttaa tcacaattgt caattttaat cctctgttta tcggcagttc
    13861 gtagagcgcg ccgtgcgtcc cgagcgatac tgagcgaagc aagtgcgtcg agcagtgccc
    13921 gcttgttcct gaaatgccag taaagcgctg gctgctgaac ccccagccgg aactgacccc
    13981 acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg
    14041 ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg
    14101 gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg
    14161 tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc
    14221 catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg
    14281 acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac
    14341 accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc
    14401 gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg
    14461 caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc
    14521 aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg
    14581 atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt
    14641 ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc
    14701 gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc
    14761 gtgtgtttca gcaacgcggc ctgcttggcc tcgctgacct gttttgccag gtcctcgccg
    14821 gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc
    14881 aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga tggcgcgggc
    14941 agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggacc
    15001 atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg
    15061 gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg
    15121 tcaaacgtcc gattcattca ccctccttgc gggattgccc cgactcacgc cggggcaatg
    15181 tgcccttatt cctgatttga cccgcctggt gccttggtgt ccagataatc caccttatcg
    15241 gcaatgaagt cggtcccgta gaccgtctgg ccgtccttct cgtacttggt attccgaatc
    15301 ttgccctgca cgaataccag cgaccccttg cccaaatact tgccgtgggc ctcggcctga
    15361 gagccaaaac acttgatgcg gaagaagtcg gtgcgctcct gcttgtcgcc ggcatcgttg
    15421 cgccacatct aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca
    15481 atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc
    15541 ctccctgatc gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct
    15601 cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag
    15661 gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag
    15721 ctcgcgcgga tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag
    15781 atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg
    15841 acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat
    15901 cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact
    15961 catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag
    16021 ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt
    16081 ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac
    16141 cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt
    16201 caattccggt gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat
    16261 ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact gttccttgca
    16321 ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat
    16381 aacatagtat cgacggagcc gattttgaaa ccacaattat gggtgatgct gccaacttac
    16441 tgatttagtg tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt
    16501 ccctcctgtt cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag
    16561 cgctatctct gctctcactg ccgtaaaaca tggcaactgc agttcactta caccgcttct
    16621 caacccggta cgcaccagaa aatcattgat atggccatga atggcgttgg atgccgggca
    16681 acagcccgca ttatgggcgt tggcctcaac acgattttac gtcacttaaa aaactcaggc
    16741 cgcagtcggt aacctcgcgc atacagccgg gcagtgacgt catcgtctgc gcggaaatgg
    16801 acgaacagtg gggctatgtc ggggctaaat cgcgccagcg ctggctgttt tacgcgtatg
    16861 acagtctccg gaagacggtt gttgcgcacg tattcggtga acgcactatg gcgacgctgg
    16921 ggcgtcttat gagcctgctg tcaccctttg acgtggtgat atggatgacg gatggctggc
    16981 cgctgtatga atcccgcctg aagggaaagc tgcacgtaat cagcaagcga tatacgcagc
    17041 gaattgagcg gcataacctg aatctgaggc agcacctggc acggctggga cggaagtcgc
    17101 tgtcgttctc aaaatcggtg gagctgcatg acaaagtcat cgggcattat ctgaacataa
    17161 aacactatca ataagttgga gtcattaccc aattatgata gaatttacaa gctataaggt
    17221 tattgtcctg ggtttcaagc attagtccat gcaagttttt atgctttgcc cattctatag
    17281 atatattgat aagcgcgctg cctatgcctt gccccctgaa atccttacat acggcgatat
    17341 cttctatata aaagatatat tatcttatca gtattgtcaa tatattcaag gcaatctgcc
    17401 tcctcatcct cttcatcctc ttcgtcttgg tagcttttta aatatggcgc ttcatagagt
    17461 aattctgtaa aggtccaatt ctcgttttca tacctcggta taatcttacc tatcacctca
    17521 aatggttcgc tgggtttatc gcacccccga acacgagcac ggcacccgcg accactatgc
    17581 caagaatgcc caaggtaaaa attgccggcc ccgccatgaa gtccgtgaat gccccgacgg
    17641 ccgaagtgaa gggcaggccg ccacccaggc cgccgccctc actgcccggc acctggtcgc
    17701 tgaatgtcga tgccagcacc tgcggcacgt caatgcttcc gggcgtcgcg ctcgggctga
    17761 tcgcccatcc cgttactgcc ccgatcccgg caatggcaag gactgccagc gctgccattt
    17821 ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt
    17881 tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg
    17941 cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa
    18001 aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg
    18061 cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc
    18121 tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac
    18181 cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag
    18241 gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc
    18301 ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct
    18361 catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccggccgcg
    18421 gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc
    18481 cagcccagcg gcgagggcaa ccagcccggt gagcgtcgga aaggcgc
//
