LOCUS       sequence-202758-       18336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101090 (Pen3). Also 
            expresses Cas9 from AtUBQ10 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18336)
  AUTHORS   Curtin SJ, Tiffin P, Guhlin J, Trujillo DI, Burghart LT, Atkins P, 
            Baltes NJ, Denny R, Voytas DF, Stupar RM, Young ND
  TITLE     Validating Genome-Wide Association Candidates Controlling 
            Quantitative Variation in Nodulation.
  JOURNAL   Plant Physiol. 2017 Feb;173(2):921-931. doi: 10.1104/pp.16.01923. 
            Epub 2017 Jan 5.
  PUBMED    28057894
REFERENCE   2  (bases 1 to 18336)
  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"; doi: 
            "10.1104/pp.16.01923"; journalName: "Plant Physiol"; date: 
            "2017-02"; volume: "173"; issue: "2"; pages: "921-931"
COMMENT     Sequence Label: pSC32
FEATURES             Location/Qualifiers
     source          1..18336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    59..83
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             105..152
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             159..179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(204..1640,1830..4493)
                     /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          1641..1829
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4494..4541
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4552..4804
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4849..4869
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5224..5299
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5311..5335)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5812..5887
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5899..5923)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5942..5966)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6030..6282
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6300..6316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6300..6316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6582..6606
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7071..7721
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7909..7926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8160..8179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8402..8511
                     /note="incP origin of transfer"
     CDS             8544..8915
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      9042..9757
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10380..11147
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11339..12133
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12432..13580
                     /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    15176..15200
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15447..15468)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15659..15678
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15818..15834
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15847..16099)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16243..16794)
                     /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(16815..17160)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16852..16872)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17696..17712
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgagtttttc tgattaacag ggtaccgggc cccccctcga ggcgcgccaa gctatcaaac
       61 aagtttgtac aaaaaagcag gctccgaatt cgcccttcac catggattac aaggatgatg
      121 atgataagga ttacaaggat gatgatgata agatggctcc aaagaagaag agaaaggttg
      181 gaatccacgg agttccagct gctgataaga agtactctat cggacttgac atcggaacca
      241 actctgttgg atgggctgtt atcaccgatg agtacaaggt tccatctaag aagttcaagg
      301 ttcttggaaa caccgataga cactctatca agaagaacct tatcggtgct cttcttttcg
      361 attctggaga gaccgctgag gctaccagat tgaagagaac cgctagaaga agatacacca
      421 gaagaaagaa cagaatctgc taccttcagg aaatcttctc taacgagatg gctaaggttg
      481 atgattcttt cttccacaga cttgaggagt ctttccttgt tgaggaggat aagaagcacg
      541 agagacaccc aatcttcgga aacatcgttg atgaggttgc ttaccacgag aagtacccaa
      601 ccatctacca ccttagaaag aagttggttg attctaccga taaggctgat cttagactta
      661 tctaccttgc tcttgctcac atgatcaagt tcagaggaca cttccttatc gagggagacc
      721 ttaacccaga taactctgat gttgataagt tgttcatcca gcttgttcag acctacaacc
      781 agcttttcga ggagaaccca atcaacgctt ctggagttga tgctaaggct atcctttctg
      841 ctagactttc taagtctcgt agacttgaga accttatcgc tcagcttcca ggagagaaga
      901 agaacggact tttcggaaac cttatcgctc tttctcttgg acttacccca aacttcaagt
      961 ctaacttcga tcttgctgag gatgctaagt tgcagctttc taaggatacc tacgatgatg
     1021 atcttgataa ccttcttgct cagatcggag atcagtacgc tgatcttttc cttgctgcta
     1081 agaacctttc tgatgctatc cttctttctg acatccttag agttaacacc gagatcacca
     1141 aggctccact ttctgcttct atgatcaaga gatacgatga gcaccaccag gatcttaccc
     1201 ttttgaaggc tcttgttaga cagcagcttc cagagaagta caaggaaatc ttcttcgatc
     1261 agtctaagaa cggatacgct ggatacatcg atggaggagc ttctcaggag gagttctaca
     1321 agttcatcaa gccaatcctt gagaagatgg atggaaccga ggagcttctt gttaagttga
     1381 acagagagga tcttcttaga aagcagagaa ccttcgataa cggatctatc ccacaccaga
     1441 tccaccttgg agagcttcac gctatccttc gtagacagga ggatttctac ccattcttga
     1501 aggataacag agagaagatc gagaagatcc ttaccttcag aatcccatac tacgttggac
     1561 cacttgctag aggaaactct cgtttcgctt ggatgaccag aaagtctgag gagaccatca
     1621 ccccttggaa cttcgaggag gtaagtttct gcttctacct ttgatatata tataataatt
     1681 atcattaatt agtagtaata taatatttca aatatttttt tcaaaataaa agaatgtagt
     1741 atatagcaat tgcttttctg tagtttataa gtgtgtatat tttaatttat aacttttcta
     1801 atatatgacc aaaatttgtt gatgtgcagg ttgttgataa gggagcttct gctcagtctt
     1861 tcatcgagag aatgaccaac ttcgataaga accttccaaa cgagaaggtt cttccaaagc
     1921 actctcttct ttacgagtac ttcaccgttt acaacgagct taccaaggtt aagtacgtta
     1981 ccgagggaat gagaaagcca gctttccttt ctggagagca gaagaaggct atcgttgatc
     2041 ttcttttcaa gaccaacaga aaggttaccg ttaagcagtt gaaggaggat tacttcaaga
     2101 agatcgagtg cttcgattct gttgaaatct ctggagttga ggatagattc aacgcttctc
     2161 ttggaaccta ccacgatctt ttgaagatca tcaaggataa ggatttcctt gataacgagg
     2221 agaacgagga catccttgag gacatcgttc ttacccttac ccttttcgag gatagagaga
     2281 tgatcgagga gagactcaag acctacgctc accttttcga tgataaggtt atgaagcagt
     2341 tgaagagaag aagatacacc ggatggggta gactttctcg taagttgatc aacggaatca
     2401 gagataagca gtctggaaag accatccttg atttcttgaa gtctgatgga ttcgctaaca
     2461 gaaacttcat gcagcttatc cacgatgatt ctcttacctt caaggaggac atccagaagg
     2521 ctcaggtttc tggacaggga gattctcttc acgagcacat cgctaacctt gctggatctc
     2581 cagctatcaa gaagggaatc cttcagaccg ttaaggttgt tgatgagctt gttaaggtta
     2641 tgggtagaca caagccagag aacatcgtta tcgagatggc tagagagaac cagaccaccc
     2701 agaagggaca gaagaactct cgtgagagaa tgaagagaat cgaggaggga atcaaggagc
     2761 ttggatctca aatcttgaag gagcacccag ttgagaacac ccagcttcag aacgagaagt
     2821 tgtaccttta ctaccttcag aacggaagag atatgtacgt tgatcaggag cttgacatca
     2881 acagactttc tgattacgat gttgatcaca tcgttccaca gtctttcttg aaggatgatt
     2941 ctatcgataa caaggttctt acccgttctg ataagaacag aggaaagtct gataacgttc
     3001 catctgagga ggttgttaag aagatgaaga actactggag acagcttctt aacgctaagt
     3061 tgatcaccca gagaaagttc gataacctta ccaaggctga gagaggagga ctttctgagc
     3121 ttgataaggc tggattcatc aagagacagc ttgttgagac cagacagatc accaagcacg
     3181 ttgctcagat ccttgattct cgtatgaaca ccaagtacga tgagaacgat aagttgatca
     3241 gagaggttaa ggttatcacc ttgaagtcta agttggtttc tgatttcaga aaggatttcc
     3301 agttctacaa ggttagagag atcaacaact accaccacgc tcacgatgct taccttaacg
     3361 ctgttgttgg aaccgctctt atcaagaagt acccaaagtt ggagtctgag ttcgtttacg
     3421 gagattacaa ggtttacgat gttagaaaga tgatcgctaa gtctgagcag gagatcggaa
     3481 aggctaccgc taagtacttc ttctactcta acatcatgaa cttcttcaag accgagatca
     3541 cccttgctaa cggagagatc agaaagagac cacttatcga gaccaacgga gagaccggag
     3601 agatcgtttg ggataaggga agagatttcg ctaccgttag aaaggttctt tctatgccac
     3661 aggttaacat cgttaagaaa accgaggttc agaccggagg attctctaag gagtctatcc
     3721 ttccaaagag aaactctgat aagttgatcg ctagaaagaa ggattgggac ccaaagaagt
     3781 acggaggatt cgattctcca accgttgctt actctgttct tgttgttgct aaggttgaga
     3841 agggaaagtc taagaagttg aagtctgtta aggagcttct tggaatcacc atcatggagc
     3901 gttcttcttt cgagaagaac ccaatcgatt tccttgaggc taagggatac aaggaggtta
     3961 agaaggatct tatcatcaag ttgccaaagt actctctttt cgagcttgag aacggaagaa
     4021 agagaatgct tgcttctgct ggagagcttc agaagggaaa cgagcttgct cttccatcta
     4081 agtacgttaa cttcctttac cttgcttctc actacgagaa gttgaaggga tctccagagg
     4141 ataacgagca gaagcagctt ttcgttgagc agcacaagca ctaccttgat gagatcatcg
     4201 agcaaatctc tgagttctct aagagagtta tccttgctga tgctaacctt gataaggttc
     4261 tttctgctta caacaagcac agagataagc caatcagaga gcaggctgag aacatcatcc
     4321 accttttcac ccttaccaac cttggtgctc cagctgcttt caagtacttc gataccacca
     4381 tcgatagaaa aagatacacc tctaccaagg aggttcttga tgctaccctt atccaccagt
     4441 ctatcaccgg actttacgag accagaatcg atctttctca gcttggagga gataagagac
     4501 cagctgctac caagaaggct ggacaggcta agaagaagaa gtgagacgtc cgatcgttca
     4561 aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc gatgattatc
     4621 atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg catgacgtta
     4681 tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata cgcgatagaa
     4741 aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta
     4801 gatcgggaat tgatcccccc tcgacagctt ccggaaaggg cgaattcgca actttgtata
     4861 caaaagttgc cgagctcgcg gccgcatgtt gttgttacca gaaagtaaat aaatgttcaa
     4921 tctctgatgt tctcaagtaa gtgagtttta ttgggaataa tattaactta tgttcttctt
     4981 gcatttgatt tctttgccgc tctcttcttc tatcttaaat ctgtgtatac tatttcacta
     5041 ttgggctttt tattagtcta taatgggact caaaataagg ctttggccca catcaaaaag
     5101 ataagtcaca aatcaaaact aaattcagag tcttttctcc cacatcggtc actgtactca
     5161 ttttgtgttt gtttatatat tacacgaacc gatctttggt acggatttct cagctagatg
     5221 ccagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
     5281 agtggcaccg agtcggtgct ttttttctag acccagcttt cttgtacaaa gttggcatta
     5341 ccatggcttc gttgaacaac ggaaactcga cttgccttcc gcacaataca tcatttcttc
     5401 ttagcttttt ttcttcttct tcgttcatac agtttttttt tgtttatcag cttacatttt
     5461 cttgaaccgt agctttcgtt ttcttctttt taactttcca ttcggagttt ttgtatcttg
     5521 tttcatagtt tgtcccagga ttagaatgat taggcatcga accttcaaga atttgattga
     5581 ataaaacatc ttcattctta agatatgaag ataatcttca aaaggcccct gggaatctga
     5641 aagaagagaa gcaggcccat ttatatggga aagaacaata gtatttctta tataggccca
     5701 tttaagttga aaacaatctt caaaagtccc acatcgctta gataagaaaa cgaagctgag
     5761 tttatataca gctagagtcg aagtagtgat tgggacttct caaagcatgt tgttttagag
     5821 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     5881 tcggtgcttt ttttctagac ccagctttct tgtacaaagt tggcattact gcaggtcgac
     5941 gacccagctt tcttgtacaa agtggttcga taattcttaa ttaactagtt ctagagcggc
     6001 cgccaccgcg gtggagctcg aatttccccg atcgttcaaa catttggcaa taaagtttct
     6061 taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg
     6121 ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga
     6181 ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact
     6241 aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcgggaattc gtaatcatgg
     6301 tcatagctgt ttcctgccta gtcggagcgg ccgcgtccgt ctagctgata gtgaccttag
     6361 gcgacttttg aacgcgcaat aatggtttct gacgtatgtg cttagctcat taaactccag
     6421 aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg
     6481 gcttgtcccg cgtcatcggc gggggtcata acgtgactcc cttaattctc cgctcatgat
     6541 cagattgtcg tttcccgcct tcagtttaaa ctatcagtgt ttgacaggat atattggcgg
     6601 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
     6661 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc agatctggcg
     6721 ccggccagcg agacgagcaa gattggccgc cgcccgaaac gatccgacag cgcgcccagc
     6781 acaggtgcgc aggcaaattg caccaacgca tacagcgcca gcagaatgcc atagtgggcg
     6841 gtgacgtcgt tcgagtgaac cagatcgcgc aggaggcccg gcagcaccgg cataatcagg
     6901 ccgatgccga cagcgtcgag cgcgacagtg ctcagaatta cgatcagggg tatgttgggt
     6961 ttcacgtctg gcctccggac cagcctccgc tggtccgatt gaacgcgcgg attctttatc
     7021 actgataagt tggtggacat attatgttta tcagtgataa agtgtcaagc atgacaaagt
     7081 tgcagccgaa tacagtgatc cgtgccgccc tggacctgtt gaacgaggtc ggcgtagacg
     7141 gtctgacgac acgcaaactg gcggaacggt tgggggttca gcagccggcg ctttactggc
     7201 acttcaggaa caagcgggcg ctgctcgacg cactggccga agccatgctg gcggagaatc
     7261 atacgcattc ggtgccgaga gccgacgacg actggcgctc atttctgatc gggaatgccc
     7321 gcagcttcag gcaggcgctg ctcgcctacc gcgatggcgc gcgcatccat gccggcacgc
     7381 gaccgggcgc accgcagatg gaaacggccg acgcgcagct tcgcttcctc tgcgaggcgg
     7441 gtttttcggc cggggacgcc gtcaatgcgc tgatgacaat cagctacttc actgttgggg
     7501 ccgtgcttga ggagcaggcc ggcgacagcg atgccggcga gcgcggcggc accgttgaac
     7561 aggctccgct ctcgccgctg ttgcgggccg cgatagacgc cttcgacgaa gccggtccgg
     7621 acgcagcgtt cgagcaggga ctcgcggtga ttgtcgatgg attggcgaaa aggaggctcg
     7681 ttgtcaggaa cgttgaagga ccgagaaagg gtgacgattg atcaggaccg ctgccggagc
     7741 gcaacccact cactacagca gagccatgta gacaacatcc cctccccctt tccaccgcgt
     7801 cagacgcccg tagcagcccg ctacgggctt tttcatgccc tgccctagcg tccaagcctc
     7861 acggccgcgc tcggcctctc tggcggcctt ctggcgctct tccgcttcct cgctcactga
     7921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     8041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     8101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     8161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     8221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt tccgctgcat
     8281 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
     8341 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
     8401 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
     8461 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg
     8521 gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc
     8581 cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg
     8641 cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg
     8701 gcgtcgtgga ctatgagcac gtccgcgagc tggcccgcat caatggcgac ctgggccgcc
     8761 tgggcggcct gctgaaactc tggctcaccg acgacccgcg cacggcgcgg ttcggtgatg
     8821 ccacgatcct cgccctgctg gcgaagatcg aagagaagca ggacgagctt ggcaaggtca
     8881 tgatgggcgt ggtccgcccg agggcagagc catgactttt ttagccgcta aaacggccgg
     8941 ggggtgcgcg tgattgccaa gcacgtcccc atgcgctcca tcaagaagag cgacttcgcg
     9001 gagctggtga agtacatcac cgacgagcaa ggcaagaccg agcgcctttc cgacgctcac
     9061 cgggctggtt gccctcgccg ctgggctggc ggccgtctat ggccctgcaa acgcgccaga
     9121 aacgccgtcg aagccgtgtg cgagacaccg cggccggccg ccggcgttgt ggatacctcg
     9181 cggaaaactt ggccctcact gacagatgag gggcggacgt tgacacttga ggggccgact
     9241 cacccggcgc ggcgttgaca gatgaggggc aggctcgatt tcggccggcg acgtggagct
     9301 ggccagcctc gcaaatcggc gaaaacgcct gattttacgc gagtttccca cagatgatgt
     9361 ggacaagcct ggggataagt gccctgcggt attgacactt gaggggcgcg actactgaca
     9421 gatgaggggc gcgatccttg acacttgagg ggcagagtgc tgacagatga ggggcgcacc
     9481 tattgacatt tgaggggctg tccacaggca gaaaatccag catttgcaag ggtttccgcc
     9541 cgtttttcgg ccaccgctaa cctgtctttt aacctgcttt taaaccaata tttataaacc
     9601 ttgtttttaa ccagggctgc gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc
     9661 ccccttctcg aaccctcccg gcccgctaac gcgggcctcc catcccccca ggggctgcgc
     9721 ccctcggccg cgaacggcct caccccaaaa atggcagcgc tggcagtcct tgccattgcc
     9781 gggatcgggg cagtaacggg atgggcgatc agcccgagcg cgacgcccgg aagcattgac
     9841 gtgccgcagg tgctggcatc gacattcagc gaccaggtgc cgggcagtga gggcggcggc
     9901 ctgggtggcg gcctgccctt cacttcggcc gtcggggcat tcacggactt catggcgggg
     9961 ccggcaattt ttaccttggg cattcttggc atagtggtcg cgggtgccgt gctcgtgttc
    10021 gggggtgcga taaacccagc gaaccatttg aggtgatagg taagattata ccgaggtatg
    10081 aaaacgagaa ttggaccttt acagaattac tctatgaagc gccatattta aaaagctacc
    10141 aagacgaaga ggatgaagag gatgaggagg cagattgcct tgaatatatt gacaatactg
    10201 ataagataat atatctttta tatagaagat atcgccgtat gtaaggattt cagggggcaa
    10261 ggcataggca gcgcgcttat caatatatct atagaatggg caaagcataa aaacttgcat
    10321 ggactaatgc ttgaaaccca ggacaataac cttatagctt gtaaattcta tcataattgg
    10381 gtaatgactc caacttattg atagtgtttt atgttcagat aatgcccgat gactttgtca
    10441 tgcagctcca ccgattttga gaacgacagc gacttccgtc ccagccgtgc caggtgctgc
    10501 ctcagattca ggttatgccg ctcaattcgc tgcgtatatc gcttgctgat tacgtgcagc
    10561 tttcccttca ggcgggattc atacagcggc cagccatccg tcatccatat caccacgtca
    10621 aagggtgaca gcaggctcat aagacgcccc agcgtcgcca tagtgcgttc accgaatacg
    10681 tgcgcaacaa ccgtcttccg gagactgtca tacgcgtaaa acagccagcg ctggcgcgat
    10741 ttagccccga catagcccca ctgttcgtcc atttccgcgc agacgatgac gtcactgccc
    10801 ggctgtatgc gcgaggttac cgactgcggc ctgagttttt taagtgacgt aaaatcgtgt
    10861 tgaggccaac gcccataatg cgggctgttg cccggcatcc aacgccattc atggccatat
    10921 caatgatttt ctggtgcgta ccgggttgag aagcggtgta agtgaactgc agttgccatg
    10981 ttttacggca gtgagagcag agatagcgct gatgtccggc ggtgcttttg ccgttacgca
    11041 ccaccccgtc agtagctgaa caggagggac agctgataga aacagaagcc actggagcac
    11101 ctcaaaaaca ccatcataca ctaaatcagt aagttggcag catcacccat aattgtggtt
    11161 tcaaaatcgg ctccgtcgat actatgttat acgccaactt tgaaaacaac tttgaaaaag
    11221 ctgttttctg gtatttaagg ttttagaatg caaggaacag tgaattggag ttcgtcttgt
    11281 tataattagc ttcttggggt atctttaaat actgtagaaa agaggaagga aataataaat
    11341 ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa
    11401 agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct
    11461 atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa
    11521 ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg
    11581 gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga
    11641 gtatgaagat gaacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct
    11701 ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc
    11761 cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga
    11821 agacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga
    11881 agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg
    11941 caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat
    12001 tgccttctgc gtccggtcga tcagggagga tatcggggaa gaacagtatg tcgagctatt
    12061 ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga
    12121 tgaattgttt tagtacctag atgtggcgca acgatgccgg cgacaagcag gagcgcaccg
    12181 acttcttccg catcaagtgt tttggctctc aggccgaggc ccacggcaag tatttgggca
    12241 aggggtcgct ggtattcgtg cagggcaaga ttcggaatac caagtacgag aaggacggcc
    12301 agacggtcta cgggaccgac ttcattgccg ataaggtgga ttatctggac accaaggcac
    12361 caggcgggtc aaatcaggaa taagggcaca ttgccccggc gtgagtcggg gcaatcccgc
    12421 aaggagggtg aatgaatcgg acgtttgacc ggaaggcata caggcaagaa ctgatcgacg
    12481 cggggttttc cgccgaggat gccgaaacca tcgcaagccg caccgtcatg cgtgcgcccc
    12541 gcgaaacctt ccagtccgtc ggctcgatgg tccagcaagc tacggccaag atcgagcgcg
    12601 acagcgtgca actggctccc cctgccctgc ccgcgccatc ggccgccgtg gagcgttcgc
    12661 gtcgtctcga acaggaggcg gcaggtttgg cgaagtcgat gaccatcgac acgcgaggaa
    12721 ctatgacgac caagaagcga aaaaccgccg gcgaggacct ggcaaaacag gtcagcgagg
    12781 ccaagcaggc cgcgttgctg aaacacacga agcagcagat caaggaaatg cagctttcct
    12841 tgttcgatat tgcgccgtgg ccggacacga tgcgagcgat gccaaacgac acggcccgct
    12901 ctgccctgtt caccacgcgc aacaagaaaa tcccgcgcga ggcgctgcaa aacaaggtca
    12961 ttttccacgt caacaaggac gtgaagatca cctacaccgg cgtcgagctg cgggccgacg
    13021 atgacgaact ggtgtggcag caggtgttgg agtacgcgaa gcgcacccct atcggcgagc
    13081 cgatcacctt cacgttctac gagctttgcc aggacctggg ctggtcgatc aatggccggt
    13141 attacacgaa ggccgaggaa tgcctgtcgc gcctacaggc gacggcgatg ggcttcacgt
    13201 ccgaccgcgt tgggcacctg gaatcggtgt cgctgctgca ccgcttccgc gtcctggacc
    13261 gtggcaagaa aacgtcccgt tgccaggtcc tgatcgacga ggaaatcgtc gtgctgtttg
    13321 ctggcgacca ctacacgaaa ttcatatggg agaagtaccg caagctgtcg ccgacggccc
    13381 gacggatgtt cgactatttc agctcgcacc gggagccgta cccgctcaag ctggaaacct
    13441 tccgcctcat gtgcggatcg gattccaccc gcgtgaagaa gtggcgcgag caggtcggcg
    13501 aagcctgcga agagttgcga ggcagcggcc tggtggaaca cgcctgggtc aatgatgacc
    13561 tggtgcattg caaacgctag ggccttgtgg ggtcagttcc ggctgggggt tcagcagcca
    13621 gcgctttact ggcatttcag gaacaagcgg gcactgctcg acgcacttgc ttcgctcagt
    13681 atcgctcggg acgcacggcg cgctctacga actgccgata aacagaggat taaaattgac
    13741 aattgtgatt aaggctcaga ttcgacggct tggagcggcc gacgtgcagg atttccgcga
    13801 gatccgattg tcggccctga agaaagctcc agagatgttc gggtccgttt acgagcacga
    13861 ggagaaaaag cccatggagg cgttcgctga acggttgcga gatgccgtgg cattcggcgc
    13921 ctacatcgac ggcgagatca ttgggctgtc ggtcttcaaa caggaggacg gccccaagga
    13981 cgctcacaag gcgcatctgt ccggcgtttt cgtggagccc gaacagcgag gccgaggggt
    14041 cgccggtatg ctgctgcggg cgttgccggc gggtttattg ctcgtgatga tcgtccgaca
    14101 gattccaacg ggaatctggt ggatgcgcat cttcatcctc ggcgcactta atatttcgct
    14161 attctggagc ttgttgttta tttcggtcta ccgcctgccg ggcggggtcg cggcgacggt
    14221 aggcgctgtg cagccgctga tggtcgtgtt catctctgcc gctctgctag gtagcccgat
    14281 acgattgatg gcggtcctgg gggctatttg cggaactgcg ggcgtggcgc tgttggtgtt
    14341 gacaccaaac gcagcgctag atcctgtcgg cgtcgcagcg ggcctggcgg gggcggtttc
    14401 catggcgttc ggaaccgtgc tgacccgcaa gtggcaacct cccgtgcctc tgctcacctt
    14461 taccgcctgg caactggcgg ccggaggact tctgctcgtt ccagtagctt tagtgtttga
    14521 tccgccaatc ccgatgccta caggaaccaa tgttctcggc ctggcgtggc tcggcctgat
    14581 cggagcgggt ttaacctact tcctttggtt ccgggggatc tcgcgactcg aacctacagt
    14641 tgtttcctta ctgggctttc tcagccccag atctggggtc gatcagccgg ggatgcatca
    14701 ggccgacagt cggaacttcg ggtccccgac ctgtaccatt cggtgagcaa tggatagggg
    14761 agttgatatc gtcaacgttc acttctaaag aaatagcgcc actcagcttc ctcagcggct
    14821 ttatccagcg atttcctatt atgtcggcat agttctcaag atcgacagcc tgtcacggtt
    14881 aagcgagaaa tgaataagaa ggctgataat tcggatctct gcgagggaga tgatatttga
    14941 tcacaggcag caacgctctg tcatcgttac aatcaacatg ctaccctccg cgagatcatc
    15001 cgtgtttcaa acccggcagc ttagttgccg ttcttccgaa tagcatcggt aacatgagca
    15061 aagtctgccg ccttacaacg gctctcccgc tgacgccgtc ccggactgat gggctgcctg
    15121 tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca
    15181 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
    15241 ttttaatgta ctggggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
    15301 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
    15361 cgaaaatcct gtttgatggt ggttccgaaa tcggcaaaat cccttataaa tcaaaagaat
    15421 agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
    15481 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
    15541 catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
    15601 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
    15661 ggaagaaagc gaaaggagcg ggcgccattc aggctgcgca actgttggga agggcgatcg
    15721 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
    15781 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatta
    15841 attcccgatc tagtaacata gatgacaccg cgcgcgataa tttatcctag tttgcgcgct
    15901 atattttgtt ttctatcgcg tattaaatgt ataattgcgg gactctaatc ataaaaaccc
    15961 atctcataaa taacgtcatg cattacatgt taattattac atgcttaacg taattcaaca
    16021 gaaattatat gataatcatc gcaagaccgg caacaggatt caatcttaag aaactttatt
    16081 gccaaatgtt tgaacgatcg gggaaattcg agctcggtac cccctcctgg gcacggtcgt
    16141 acgcgatccg ccgcggttcc acctcaccat tgcgggtatg ccagttgagt tcgatgagtt
    16201 cccgtaccag ctccaactcg ggcgatgcca cgcgcactcc gttcagatct cggtgacggg
    16261 caggaccgga cggggcggta ccggcaggct gaagtccagc tgccagaaac ccacgtcatg
    16321 ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc
    16381 ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa
    16441 gccctgtgcc tccagggact tcagcaggtg ggtgtagagc gtggagccca gtcccgtccg
    16501 ctggtggcgg ggggagacgt acacggtcga ctcggccgtc cagtcgtagg cgttgcgtgc
    16561 cttccagggg cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca
    16621 gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc
    16681 ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg
    16741 catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcatggatcc
    16801 tctagagtcc cccgtgttct ctccaaatga aatgaacttc cttatataga ggaagggtct
    16861 tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc acatcaatcc
    16921 acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct cgtgggtggg
    16981 ggtccatctt tgggaccact gtcggcagag gcatcttcaa cgatggcctt tcctttatcg
    17041 caatgatggc atttgtagga gccaccttcc ttttccacta tcttcacaat aaagtgacag
    17101 atagctgggc aatggaatcc gaggaggttt ccggatatta ccctttgttg aaaagtctca
    17161 attgcccttt ggtcttctga gactgtatct ttgatatttt tggagtagac aagtgtgtcg
    17221 tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaagag actctgtatg
    17281 aactgttcgc cagtctttac ggcgagttct gttaggtcct ctatttgaat ctttgactcc
    17341 atggcctttg attcagtggg aactaccttt ttagagactc caatctctat tacttgcctt
    17401 ggtttgtgaa gcaagccttg aatcgtccat actggaatag tacttctgat cttgagaaat
    17461 atatctttct ctgtgttctt gatgcagtta gtcctgaatc ttttgactgc atctttaacc
    17521 ttcttgggaa ggtatttgat ttcctggaga ttattgctcg ggtagatcgt cttgatgaga
    17581 cctgctgcgt aagcctctct aaccatctgt gggttagcat tctttctgaa attgaaaagg
    17641 ctaatctggg gacctgcagg ggcaataccg ccagggtttt cccagtcacg acgttgtaaa
    17701 acgacggcca gtgccaagct ttcgacgagt cagtaataaa cggcgtcaaa gtggttgcag
    17761 ccggcacaca cgagtcgtgt ttatcaactc aaagcacaaa tacttttcct caacctaaaa
    17821 ataaggcaat tagccaaaaa caactttgcg tgtaaacaac gctcaataca cgtgtcattt
    17881 tattattagc tattgcttca ccgccttagc tttctcgtga cctagtcgtc ctcgtctttt
    17941 cttcttcttc ttctataaaa caatacccaa agagctcttc ttcttcacaa ttcagatttc
    18001 aatttctcaa aatcttaaaa actttctctc aattctctct accgtgatca aggtaaattt
    18061 ctgtgttcct tattctctca aaatcttcga ttttgttttc gttcgatccc aatttcgtat
    18121 atgttctttg gtttagattc tgttaatctt agatcgaaga cgattttctg ggtttgatcg
    18181 ttagatatca tcttaattct cgattagggt ttcatagata tcatccgatt tgttcaaata
    18241 atttgagttt tgtcgaataa ttactcttcg atttgtgatt tctatctaga tctggtgtta
    18301 gtttctagtt tgtgcgatcg aatttgtaga ttaatc
//
