LOCUS       sequence-203134-       18337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Glyma17g11235 (Dcl4a). Also 
            expresses Cas9 from AtUBQ10 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18337)
  AUTHORS   Shaun Curtin
  TITLE     Shaun Curtin CRISPR plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 18337)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSC49
FEATURES             Location/Qualifiers
     source          1..18337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    30..54
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             76..123
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             130..150
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(175..1611,1801..4464)
                     /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          1612..1800
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4465..4512
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4523..4775
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4820..4840
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5195..5270
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5282..5306)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5784..5859
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5871..5895)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5914..5938)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6002..6254
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6272..6288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6272..6288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6554..6578
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7043..7693
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7881..7898)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8132..8151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8374..8483
                     /note="incP origin of transfer"
     CDS             8516..8887
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      9014..9729
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10352..11119
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11311..12105
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12404..13552
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    15148..15172
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15419..15440)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15631..15650
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15790..15806
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15819..16071)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16215..16766)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(16787..17132)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16824..16844)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17668..17684
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccccccctcg aggcgcgcca agctatcaaa caagtttgta caaaaaagca ggctccgaat
       61 tcgcccttca ccatggatta caaggatgat gatgataagg attacaagga tgatgatgat
      121 aagatggctc caaagaagaa gagaaaggtt ggaatccacg gagttccagc tgctgataag
      181 aagtactcta tcggacttga catcggaacc aactctgttg gatgggctgt tatcaccgat
      241 gagtacaagg ttccatctaa gaagttcaag gttcttggaa acaccgatag acactctatc
      301 aagaagaacc ttatcggtgc tcttcttttc gattctggag agaccgctga ggctaccaga
      361 ttgaagagaa ccgctagaag aagatacacc agaagaaaga acagaatctg ctaccttcag
      421 gaaatcttct ctaacgagat ggctaaggtt gatgattctt tcttccacag acttgaggag
      481 tctttccttg ttgaggagga taagaagcac gagagacacc caatcttcgg aaacatcgtt
      541 gatgaggttg cttaccacga gaagtaccca accatctacc accttagaaa gaagttggtt
      601 gattctaccg ataaggctga tcttagactt atctaccttg ctcttgctca catgatcaag
      661 ttcagaggac acttccttat cgagggagac cttaacccag ataactctga tgttgataag
      721 ttgttcatcc agcttgttca gacctacaac cagcttttcg aggagaaccc aatcaacgct
      781 tctggagttg atgctaaggc tatcctttct gctagacttt ctaagtctcg tagacttgag
      841 aaccttatcg ctcagcttcc aggagagaag aagaacggac ttttcggaaa ccttatcgct
      901 ctttctcttg gacttacccc aaacttcaag tctaacttcg atcttgctga ggatgctaag
      961 ttgcagcttt ctaaggatac ctacgatgat gatcttgata accttcttgc tcagatcgga
     1021 gatcagtacg ctgatctttt ccttgctgct aagaaccttt ctgatgctat ccttctttct
     1081 gacatcctta gagttaacac cgagatcacc aaggctccac tttctgcttc tatgatcaag
     1141 agatacgatg agcaccacca ggatcttacc cttttgaagg ctcttgttag acagcagctt
     1201 ccagagaagt acaaggaaat cttcttcgat cagtctaaga acggatacgc tggatacatc
     1261 gatggaggag cttctcagga ggagttctac aagttcatca agccaatcct tgagaagatg
     1321 gatggaaccg aggagcttct tgttaagttg aacagagagg atcttcttag aaagcagaga
     1381 accttcgata acggatctat cccacaccag atccaccttg gagagcttca cgctatcctt
     1441 cgtagacagg aggatttcta cccattcttg aaggataaca gagagaagat cgagaagatc
     1501 cttaccttca gaatcccata ctacgttgga ccacttgcta gaggaaactc tcgtttcgct
     1561 tggatgacca gaaagtctga ggagaccatc accccttgga acttcgagga ggtaagtttc
     1621 tgcttctacc tttgatatat atataataat tatcattaat tagtagtaat ataatatttc
     1681 aaatattttt ttcaaaataa aagaatgtag tatatagcaa ttgcttttct gtagtttata
     1741 agtgtgtata ttttaattta taacttttct aatatatgac caaaatttgt tgatgtgcag
     1801 gttgttgata agggagcttc tgctcagtct ttcatcgaga gaatgaccaa cttcgataag
     1861 aaccttccaa acgagaaggt tcttccaaag cactctcttc tttacgagta cttcaccgtt
     1921 tacaacgagc ttaccaaggt taagtacgtt accgagggaa tgagaaagcc agctttcctt
     1981 tctggagagc agaagaaggc tatcgttgat cttcttttca agaccaacag aaaggttacc
     2041 gttaagcagt tgaaggagga ttacttcaag aagatcgagt gcttcgattc tgttgaaatc
     2101 tctggagttg aggatagatt caacgcttct cttggaacct accacgatct tttgaagatc
     2161 atcaaggata aggatttcct tgataacgag gagaacgagg acatccttga ggacatcgtt
     2221 cttaccctta cccttttcga ggatagagag atgatcgagg agagactcaa gacctacgct
     2281 caccttttcg atgataaggt tatgaagcag ttgaagagaa gaagatacac cggatggggt
     2341 agactttctc gtaagttgat caacggaatc agagataagc agtctggaaa gaccatcctt
     2401 gatttcttga agtctgatgg attcgctaac agaaacttca tgcagcttat ccacgatgat
     2461 tctcttacct tcaaggagga catccagaag gctcaggttt ctggacaggg agattctctt
     2521 cacgagcaca tcgctaacct tgctggatct ccagctatca agaagggaat ccttcagacc
     2581 gttaaggttg ttgatgagct tgttaaggtt atgggtagac acaagccaga gaacatcgtt
     2641 atcgagatgg ctagagagaa ccagaccacc cagaagggac agaagaactc tcgtgagaga
     2701 atgaagagaa tcgaggaggg aatcaaggag cttggatctc aaatcttgaa ggagcaccca
     2761 gttgagaaca cccagcttca gaacgagaag ttgtaccttt actaccttca gaacggaaga
     2821 gatatgtacg ttgatcagga gcttgacatc aacagacttt ctgattacga tgttgatcac
     2881 atcgttccac agtctttctt gaaggatgat tctatcgata acaaggttct tacccgttct
     2941 gataagaaca gaggaaagtc tgataacgtt ccatctgagg aggttgttaa gaagatgaag
     3001 aactactgga gacagcttct taacgctaag ttgatcaccc agagaaagtt cgataacctt
     3061 accaaggctg agagaggagg actttctgag cttgataagg ctggattcat caagagacag
     3121 cttgttgaga ccagacagat caccaagcac gttgctcaga tccttgattc tcgtatgaac
     3181 accaagtacg atgagaacga taagttgatc agagaggtta aggttatcac cttgaagtct
     3241 aagttggttt ctgatttcag aaaggatttc cagttctaca aggttagaga gatcaacaac
     3301 taccaccacg ctcacgatgc ttaccttaac gctgttgttg gaaccgctct tatcaagaag
     3361 tacccaaagt tggagtctga gttcgtttac ggagattaca aggtttacga tgttagaaag
     3421 atgatcgcta agtctgagca ggagatcgga aaggctaccg ctaagtactt cttctactct
     3481 aacatcatga acttcttcaa gaccgagatc acccttgcta acggagagat cagaaagaga
     3541 ccacttatcg agaccaacgg agagaccgga gagatcgttt gggataaggg aagagatttc
     3601 gctaccgtta gaaaggttct ttctatgcca caggttaaca tcgttaagaa aaccgaggtt
     3661 cagaccggag gattctctaa ggagtctatc cttccaaaga gaaactctga taagttgatc
     3721 gctagaaaga aggattggga cccaaagaag tacggaggat tcgattctcc aaccgttgct
     3781 tactctgttc ttgttgttgc taaggttgag aagggaaagt ctaagaagtt gaagtctgtt
     3841 aaggagcttc ttggaatcac catcatggag cgttcttctt tcgagaagaa cccaatcgat
     3901 ttccttgagg ctaagggata caaggaggtt aagaaggatc ttatcatcaa gttgccaaag
     3961 tactctcttt tcgagcttga gaacggaaga aagagaatgc ttgcttctgc tggagagctt
     4021 cagaagggaa acgagcttgc tcttccatct aagtacgtta acttccttta ccttgcttct
     4081 cactacgaga agttgaaggg atctccagag gataacgagc agaagcagct tttcgttgag
     4141 cagcacaagc actaccttga tgagatcatc gagcaaatct ctgagttctc taagagagtt
     4201 atccttgctg atgctaacct tgataaggtt ctttctgctt acaacaagca cagagataag
     4261 ccaatcagag agcaggctga gaacatcatc caccttttca cccttaccaa ccttggtgct
     4321 ccagctgctt tcaagtactt cgataccacc atcgatagaa aaagatacac ctctaccaag
     4381 gaggttcttg atgctaccct tatccaccag tctatcaccg gactttacga gaccagaatc
     4441 gatctttctc agcttggagg agataagaga ccagctgcta ccaagaaggc tggacaggct
     4501 aagaagaaga agtgagacgt ccgatcgttc aaacatttgg caataaagtt tcttaagatt
     4561 gaatcctgtt gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca
     4621 tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt
     4681 cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa
     4741 attatcgcgc gcggtgtcat ctatgttact agatcgggaa ttgatccccc ctcgacagct
     4801 tccggaaagg gcgaattcgc aactttgtat acaaaagttg ccgagctcgc ggccgcatgt
     4861 tgttgttacc agaaagtaaa taaatgttca atctctgatg ttctcaagta agtgagtttt
     4921 attgggaata atattaactt atgttcttct tgcatttgat ttctttgccg ctctcttctt
     4981 ctatcttaaa tctgtgtata ctatttcact attgggcttt ttattagtct ataatgggac
     5041 tcaaaataag gctttggccc acatcaaaaa gataagtcac aaatcaaaac taaattcaga
     5101 gtcttttctc ccacatcggt cactgtactc attttgtgtt tgtttatata ttacacgaac
     5161 cgatctttgg tacggatgtg gttgaagctc tggtgtttta gagctagaaa tagcaagtta
     5221 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttcta
     5281 gacccagctt tcttgtacaa agttggcatt accatggctt cgttgaacaa cggaaactcg
     5341 acttgccttc cgcacaatac atcatttctt cttagctttt tttcttcttc ttcgttcata
     5401 cagttttttt ttgtttatca gcttacattt tcttgaaccg tagctttcgt tttcttcttt
     5461 ttaactttcc attcggagtt tttgtatctt gtttcatagt ttgtcccagg attagaatga
     5521 ttaggcatcg aaccttcaag aatttgattg aataaaacat cttcattctt aagatatgaa
     5581 gataatcttc aaaaggcccc tgggaatctg aaagaagaga agcaggccca tttatatggg
     5641 aaagaacaat agtatttctt atataggccc atttaagttg aaaacaatct tcaaaagtcc
     5701 cacatcgctt agataagaaa acgaagctga gtttatatac agctagagtc gaagtagtga
     5761 ttgtgattgg agacgcttaa ggagttttag agctagaaat agcaagttaa aataaggcta
     5821 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctag acccagcttt
     5881 cttgtacaaa gttggcatta ctgcaggtcg acgacccagc tttcttgtac aaagtggttc
     5941 gataattctt aattaactag ttctagagcg gccgccaccg cggtggagct cgaatttccc
     6001 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     6061 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     6121 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     6181 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     6241 tatgttacta gatcgggaat tcgtaatcat ggtcatagct gtttcctgcc tagtcggagc
     6301 ggccgcgtcc gtctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt
     6361 ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctgagtgg ctccttcaac
     6421 gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca
     6481 taacgtgact cccttaattc tccgctcatg atcagattgt cgtttcccgc cttcagttta
     6541 aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
     6601 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     6661 catgccaacc acagggttcc ccagatctgg cgccggccag cgagacgagc aagattggcc
     6721 gccgcccgaa acgatccgac agcgcgccca gcacaggtgc gcaggcaaat tgcaccaacg
     6781 catacagcgc cagcagaatg ccatagtggg cggtgacgtc gttcgagtga accagatcgc
     6841 gcaggaggcc cggcagcacc ggcataatca ggccgatgcc gacagcgtcg agcgcgacag
     6901 tgctcagaat tacgatcagg ggtatgttgg gtttcacgtc tggcctccgg accagcctcc
     6961 gctggtccga ttgaacgcgc ggattcttta tcactgataa gttggtggac atattatgtt
     7021 tatcagtgat aaagtgtcaa gcatgacaaa gttgcagccg aatacagtga tccgtgccgc
     7081 cctggacctg ttgaacgagg tcggcgtaga cggtctgacg acacgcaaac tggcggaacg
     7141 gttgggggtt cagcagccgg cgctttactg gcacttcagg aacaagcggg cgctgctcga
     7201 cgcactggcc gaagccatgc tggcggagaa tcatacgcat tcggtgccga gagccgacga
     7261 cgactggcgc tcatttctga tcgggaatgc ccgcagcttc aggcaggcgc tgctcgccta
     7321 ccgcgatggc gcgcgcatcc atgccggcac gcgaccgggc gcaccgcaga tggaaacggc
     7381 cgacgcgcag cttcgcttcc tctgcgaggc gggtttttcg gccggggacg ccgtcaatgc
     7441 gctgatgaca atcagctact tcactgttgg ggccgtgctt gaggagcagg ccggcgacag
     7501 cgatgccggc gagcgcggcg gcaccgttga acaggctccg ctctcgccgc tgttgcgggc
     7561 cgcgatagac gccttcgacg aagccggtcc ggacgcagcg ttcgagcagg gactcgcggt
     7621 gattgtcgat ggattggcga aaaggaggct cgttgtcagg aacgttgaag gaccgagaaa
     7681 gggtgacgat tgatcaggac cgctgccgga gcgcaaccca ctcactacag cagagccatg
     7741 tagacaacat cccctccccc tttccaccgc gtcagacgcc cgtagcagcc cgctacgggc
     7801 tttttcatgc cctgccctag cgtccaagcc tcacggccgc gctcggcctc tctggcggcc
     7861 ttctggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7921 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7981 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8221 ccttcgggaa gcgtggcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat
     8281 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
     8341 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
     8401 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
     8461 tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc
     8521 tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga
     8581 cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct
     8641 gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga
     8701 gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac
     8761 cgacgacccg cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat
     8821 cgaagagaag caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga
     8881 gccatgactt ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc
     8941 ccatgcgctc catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc
     9001 aaggcaagac cgagcgcctt tccgacgctc accgggctgg ttgccctcgc cgctgggctg
     9061 gcggccgtct atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac
     9121 cgcggccggc cgccggcgtt gtggatacct cgcggaaaac ttggccctca ctgacagatg
     9181 aggggcggac gttgacactt gaggggccga ctcacccggc gcggcgttga cagatgaggg
     9241 gcaggctcga tttcggccgg cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc
     9301 ctgattttac gcgagtttcc cacagatgat gtggacaagc ctggggataa gtgccctgcg
     9361 gtattgacac ttgaggggcg cgactactga cagatgaggg gcgcgatcct tgacacttga
     9421 ggggcagagt gctgacagat gaggggcgca cctattgaca tttgaggggc tgtccacagg
     9481 cagaaaatcc agcatttgca agggtttccg cccgtttttc ggccaccgct aacctgtctt
     9541 ttaacctgct tttaaaccaa tatttataaa ccttgttttt aaccagggct gcgccctgtg
     9601 cgcgtgaccg cgcacgccga aggggggtgc ccccccttct cgaaccctcc cggcccgcta
     9661 acgcgggcct cccatccccc caggggctgc gcccctcggc cgcgaacggc ctcaccccaa
     9721 aaatggcagc gctggcagtc cttgccattg ccgggatcgg ggcagtaacg ggatgggcga
     9781 tcagcccgag cgcgacgccc ggaagcattg acgtgccgca ggtgctggca tcgacattca
     9841 gcgaccaggt gccgggcagt gagggcggcg gcctgggtgg cggcctgccc ttcacttcgg
     9901 ccgtcggggc attcacggac ttcatggcgg ggccggcaat ttttaccttg ggcattcttg
     9961 gcatagtggt cgcgggtgcc gtgctcgtgt tcgggggtgc gataaaccca gcgaaccatt
    10021 tgaggtgata ggtaagatta taccgaggta tgaaaacgag aattggacct ttacagaatt
    10081 actctatgaa gcgccatatt taaaaagcta ccaagacgaa gaggatgaag aggatgagga
    10141 ggcagattgc cttgaatata ttgacaatac tgataagata atatatcttt tatatagaag
    10201 atatcgccgt atgtaaggat ttcagggggc aaggcatagg cagcgcgctt atcaatatat
    10261 ctatagaatg ggcaaagcat aaaaacttgc atggactaat gcttgaaacc caggacaata
    10321 accttatagc ttgtaaattc tatcataatt gggtaatgac tccaacttat tgatagtgtt
    10381 ttatgttcag ataatgcccg atgactttgt catgcagctc caccgatttt gagaacgaca
    10441 gcgacttccg tcccagccgt gccaggtgct gcctcagatt caggttatgc cgctcaattc
    10501 gctgcgtata tcgcttgctg attacgtgca gctttccctt caggcgggat tcatacagcg
    10561 gccagccatc cgtcatccat atcaccacgt caaagggtga cagcaggctc ataagacgcc
    10621 ccagcgtcgc catagtgcgt tcaccgaata cgtgcgcaac aaccgtcttc cggagactgt
    10681 catacgcgta aaacagccag cgctggcgcg atttagcccc gacatagccc cactgttcgt
    10741 ccatttccgc gcagacgatg acgtcactgc ccggctgtat gcgcgaggtt accgactgcg
    10801 gcctgagttt tttaagtgac gtaaaatcgt gttgaggcca acgcccataa tgcgggctgt
    10861 tgcccggcat ccaacgccat tcatggccat atcaatgatt ttctggtgcg taccgggttg
    10921 agaagcggtg taagtgaact gcagttgcca tgttttacgg cagtgagagc agagatagcg
    10981 ctgatgtccg gcggtgcttt tgccgttacg caccaccccg tcagtagctg aacaggaggg
    11041 acagctgata gaaacagaag ccactggagc acctcaaaaa caccatcata cactaaatca
    11101 gtaagttggc agcatcaccc ataattgtgg tttcaaaatc ggctccgtcg atactatgtt
    11161 atacgccaac tttgaaaaca actttgaaaa agctgttttc tggtatttaa ggttttagaa
    11221 tgcaaggaac agtgaattgg agttcgtctt gttataatta gcttcttggg gtatctttaa
    11281 atactgtaga aaagaggaag gaaataataa atggctaaaa tgagaatatc accggaattg
    11341 aaaaaactga tcgaaaaata ccgctgcgta aaagatacgg aaggaatgtc tcctgctaag
    11401 gtatataagc tggtgggaga aaatgaaaac ctatatttaa aaatgacgga cagccggtat
    11461 aaagggacca cctatgatgt ggaacgggaa aaggacatga tgctatggct ggaaggaaag
    11521 ctgcctgttc caaaggtcct gcactttgaa cggcatgatg gctggagcaa tctgctcatg
    11581 agtgaggccg atggcgtcct ttgctcggaa gagtatgaag atgaacaaag ccctgaaaag
    11641 attatcgagc tgtatgcgga gtgcatcagg ctctttcact ccatcgacat atcggattgt
    11701 ccctatacga atagcttaga cagccgctta gccgaattgg attacttact gaataacgat
    11761 ctggccgatg tggattgcga aaactgggaa gaagacactc catttaaaga tccgcgcgag
    11821 ctgtatgatt ttttaaagac ggaaaagccc gaagaggaac ttgtcttttc ccacggcgac
    11881 ctgggagaca gcaacatctt tgtgaaagat ggcaaagtaa gtggctttat tgatcttggg
    11941 agaagcggca gggcggacaa gtggtatgac attgccttct gcgtccggtc gatcagggag
    12001 gatatcgggg aagaacagta tgtcgagcta ttttttgact tactggggat caagcctgat
    12061 tgggagaaaa taaaatatta tattttactg gatgaattgt tttagtacct agatgtggcg
    12121 caacgatgcc ggcgacaagc aggagcgcac cgacttcttc cgcatcaagt gttttggctc
    12181 tcaggccgag gcccacggca agtatttggg caaggggtcg ctggtattcg tgcagggcaa
    12241 gattcggaat accaagtacg agaaggacgg ccagacggtc tacgggaccg acttcattgc
    12301 cgataaggtg gattatctgg acaccaaggc accaggcggg tcaaatcagg aataagggca
    12361 cattgccccg gcgtgagtcg gggcaatccc gcaaggaggg tgaatgaatc ggacgtttga
    12421 ccggaaggca tacaggcaag aactgatcga cgcggggttt tccgccgagg atgccgaaac
    12481 catcgcaagc cgcaccgtca tgcgtgcgcc ccgcgaaacc ttccagtccg tcggctcgat
    12541 ggtccagcaa gctacggcca agatcgagcg cgacagcgtg caactggctc cccctgccct
    12601 gcccgcgcca tcggccgccg tggagcgttc gcgtcgtctc gaacaggagg cggcaggttt
    12661 ggcgaagtcg atgaccatcg acacgcgagg aactatgacg accaagaagc gaaaaaccgc
    12721 cggcgaggac ctggcaaaac aggtcagcga ggccaagcag gccgcgttgc tgaaacacac
    12781 gaagcagcag atcaaggaaa tgcagctttc cttgttcgat attgcgccgt ggccggacac
    12841 gatgcgagcg atgccaaacg acacggcccg ctctgccctg ttcaccacgc gcaacaagaa
    12901 aatcccgcgc gaggcgctgc aaaacaaggt cattttccac gtcaacaagg acgtgaagat
    12961 cacctacacc ggcgtcgagc tgcgggccga cgatgacgaa ctggtgtggc agcaggtgtt
    13021 ggagtacgcg aagcgcaccc ctatcggcga gccgatcacc ttcacgttct acgagctttg
    13081 ccaggacctg ggctggtcga tcaatggccg gtattacacg aaggccgagg aatgcctgtc
    13141 gcgcctacag gcgacggcga tgggcttcac gtccgaccgc gttgggcacc tggaatcggt
    13201 gtcgctgctg caccgcttcc gcgtcctgga ccgtggcaag aaaacgtccc gttgccaggt
    13261 cctgatcgac gaggaaatcg tcgtgctgtt tgctggcgac cactacacga aattcatatg
    13321 ggagaagtac cgcaagctgt cgccgacggc ccgacggatg ttcgactatt tcagctcgca
    13381 ccgggagccg tacccgctca agctggaaac cttccgcctc atgtgcggat cggattccac
    13441 ccgcgtgaag aagtggcgcg agcaggtcgg cgaagcctgc gaagagttgc gaggcagcgg
    13501 cctggtggaa cacgcctggg tcaatgatga cctggtgcat tgcaaacgct agggccttgt
    13561 ggggtcagtt ccggctgggg gttcagcagc cagcgcttta ctggcatttc aggaacaagc
    13621 gggcactgct cgacgcactt gcttcgctca gtatcgctcg ggacgcacgg cgcgctctac
    13681 gaactgccga taaacagagg attaaaattg acaattgtga ttaaggctca gattcgacgg
    13741 cttggagcgg ccgacgtgca ggatttccgc gagatccgat tgtcggccct gaagaaagct
    13801 ccagagatgt tcgggtccgt ttacgagcac gaggagaaaa agcccatgga ggcgttcgct
    13861 gaacggttgc gagatgccgt ggcattcggc gcctacatcg acggcgagat cattgggctg
    13921 tcggtcttca aacaggagga cggccccaag gacgctcaca aggcgcatct gtccggcgtt
    13981 ttcgtggagc ccgaacagcg aggccgaggg gtcgccggta tgctgctgcg ggcgttgccg
    14041 gcgggtttat tgctcgtgat gatcgtccga cagattccaa cgggaatctg gtggatgcgc
    14101 atcttcatcc tcggcgcact taatatttcg ctattctgga gcttgttgtt tatttcggtc
    14161 taccgcctgc cgggcggggt cgcggcgacg gtaggcgctg tgcagccgct gatggtcgtg
    14221 ttcatctctg ccgctctgct aggtagcccg atacgattga tggcggtcct gggggctatt
    14281 tgcggaactg cgggcgtggc gctgttggtg ttgacaccaa acgcagcgct agatcctgtc
    14341 ggcgtcgcag cgggcctggc gggggcggtt tccatggcgt tcggaaccgt gctgacccgc
    14401 aagtggcaac ctcccgtgcc tctgctcacc tttaccgcct ggcaactggc ggccggagga
    14461 cttctgctcg ttccagtagc tttagtgttt gatccgccaa tcccgatgcc tacaggaacc
    14521 aatgttctcg gcctggcgtg gctcggcctg atcggagcgg gtttaaccta cttcctttgg
    14581 ttccggggga tctcgcgact cgaacctaca gttgtttcct tactgggctt tctcagcccc
    14641 agatctgggg tcgatcagcc ggggatgcat caggccgaca gtcggaactt cgggtccccg
    14701 acctgtacca ttcggtgagc aatggatagg ggagttgata tcgtcaacgt tcacttctaa
    14761 agaaatagcg ccactcagct tcctcagcgg ctttatccag cgatttccta ttatgtcggc
    14821 atagttctca agatcgacag cctgtcacgg ttaagcgaga aatgaataag aaggctgata
    14881 attcggatct ctgcgaggga gatgatattt gatcacaggc agcaacgctc tgtcatcgtt
    14941 acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc
    15001 cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc
    15061 gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg
    15121 ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac
    15181 gcttagacaa cttaataaca cattgcggac gtttttaatg tactggggtg gtttttcttt
    15241 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
    15301 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttccga
    15361 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga gtgttgttcc
    15421 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
    15481 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
    15541 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
    15601 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgccat
    15661 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
    15721 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
    15781 cacgacgttg taaaacgacg gccagtgaat taattcccga tctagtaaca tagatgacac
    15841 cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg cgtattaaat
    15901 gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca tgcattacat
    15961 gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca tcgcaagacc
    16021 ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat cggggaaatt
    16081 cgagctcggt accccctcct gggcacggtc gtacgcgatc cgccgcggtt ccacctcacc
    16141 attgcgggta tgccagttga gttcgatgag ttcccgtacc agctccaact cgggcgatgc
    16201 cacgcgcact ccgttcagat ctcggtgacg ggcaggaccg gacggggcgg taccggcagg
    16261 ctgaagtcca gctgccagaa acccacgtca tgccagttcc cgtgcttgaa gccggccgcc
    16321 cgcagcatgc cgcggggggc atatccgagc gcctcgtgca tgcgcacgct cgggtcgttg
    16381 ggcagcccga tgacagcgac cacgctcttg aagccctgtg cctccaggga cttcagcagg
    16441 tgggtgtaga gcgtggagcc cagtcccgtc cgctggtggc ggggggagac gtacacggtc
    16501 gactcggccg tccagtcgta ggcgttgcgt gccttccagg ggcccgcgta ggcgatgccg
    16561 gcgacctcgc cgtccacctc ggcgacgagc cagggatagc gctcccgcag acggacgagg
    16621 tcgtccgtcc actcctgcgg ttcctgcggc tcggtacgga agttgaccgt gcttgtctcg
    16681 atgtagtggt tgacgatggt gcagaccgcc ggcatgtccg cctcggtggc acggcggatg
    16741 tcggccgggc gtcgttctgg gctcatggat cctctagagt cccccgtgtt ctctccaaat
    16801 gaaatgaact tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat
    16861 cccttacgtc agtggagata tcacatcaat ccacttgctt tgaagacgtg gttggaacgt
    16921 cttctttttc cacgatgctc ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag
    16981 aggcatcttc aacgatggcc tttcctttat cgcaatgatg gcatttgtag gagccacctt
    17041 ccttttccac tatcttcaca ataaagtgac agatagctgg gcaatggaat ccgaggaggt
    17101 ttccggatat taccctttgt tgaaaagtct caattgccct ttggtcttct gagactgtat
    17161 ctttgatatt tttggagtag acaagtgtgt cgtgctccac catgttgacg aagattttct
    17221 tcttgtcatt gagtcgtaag agactctgta tgaactgttc gccagtcttt acggcgagtt
    17281 ctgttaggtc ctctatttga atctttgact ccatggcctt tgattcagtg ggaactacct
    17341 ttttagagac tccaatctct attacttgcc ttggtttgtg aagcaagcct tgaatcgtcc
    17401 atactggaat agtacttctg atcttgagaa atatatcttt ctctgtgttc ttgatgcagt
    17461 tagtcctgaa tcttttgact gcatctttaa ccttcttggg aaggtatttg atttcctgga
    17521 gattattgct cgggtagatc gtcttgatga gacctgctgc gtaagcctct ctaaccatct
    17581 gtgggttagc attctttctg aaattgaaaa ggctaatctg gggacctgca ggggcaatac
    17641 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctttcgacga
    17701 gtcagtaata aacggcgtca aagtggttgc agccggcaca cacgagtcgt gtttatcaac
    17761 tcaaagcaca aatacttttc ctcaacctaa aaataaggca attagccaaa aacaactttg
    17821 cgtgtaaaca acgctcaata cacgtgtcat tttattatta gctattgctt caccgcctta
    17881 gctttctcgt gacctagtcg tcctcgtctt ttcttcttct tcttctataa aacaataccc
    17941 aaagagctct tcttcttcac aattcagatt tcaatttctc aaaatcttaa aaactttctc
    18001 tcaattctct ctaccgtgat caaggtaaat ttctgtgttc cttattctct caaaatcttc
    18061 gattttgttt tcgttcgatc ccaatttcgt atatgttctt tggtttagat tctgttaatc
    18121 ttagatcgaa gacgattttc tgggtttgat cgttagatat catcttaatt ctcgattagg
    18181 gtttcataga tatcatccga tttgttcaaa taatttgagt tttgtcgaat aattactctt
    18241 cgatttgtga tttctatcta gatctggtgt tagtttctag tttgtgcgat cgaatttgta
    18301 gattaatctg agtttttctg attaacaggg taccggg
//
