LOCUS       sequence-202755-       18988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101090 (Pen3). Also 
            expresses Cas9 from Gmubi promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18988)
  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 18988)
  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: pSC31
FEATURES             Location/Qualifiers
     source          1..18988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(81..452)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            485..594
                     /note="incP origin of transfer"
     primer_bind     817..836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1070..1087
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1275..1925)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2390..2414
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3622..3646
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3668..3715
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3722..3742
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3767..5203,5393..8056)
                     /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          5204..5392
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8057..8104
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8115..8367
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8412..8432
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8787..8862
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8874..8898)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9375..9450
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9462..9486)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9505..9529)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9567..10274
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10824..11169
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11112..11132
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11190..11741
                     /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      11885..12137
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12150..12167)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12150..12166)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12159..12181)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12306..12325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12516..12537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12784..12808
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14404..15552)
                     /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(15851..16645)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16837..17604
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18227..18942
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 agtcgctctt cttgatggag cgcatgggga cgtgcttggc aatcacgcgc accccccggc
       61 cgttttagcg gctaaaaaag tcatggctct gccctcgggc ggaccacgcc catcatgacc
      121 ttgccaagct cgtcctgctt ctcttcgatc ttcgccagca gggcgaggat cgtggcatca
      181 ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg
      241 cccaggtcgc cattgatgcg ggccagctcg cggacgtgct catagtccac gacgcccgtg
      301 attttgtagc cctggccgac ggccagcagg taggccgaca ggctcatgcc ggccgccgcc
      361 gccttttcct caatcgctct tcgttcgtct ggaaggcagt acaccttgat aggtgggctg
      421 cccttcctgg ttggcttggt ttcatcagcc atccgcttgc cctcatctgt tacgccggcg
      481 gtagccggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
      541 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
      601 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
      661 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
      721 gcggaaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      781 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      841 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      901 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      961 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1021 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1081 gagcgaggaa gcggaagagc gccagaaggc cgccagagag gccgagcgcg gccgtgaggc
     1141 ttggacgcta gggcagggca tgaaaaagcc cgtagcgggc tgctacgggc gtctgacgcg
     1201 gtggaaaggg ggaggggatg ttgtctacat ggctctgctg tagtgagtgg gttgcgctcc
     1261 ggcagcggtc ctgatcaatc gtcacccttt ctcggtcctt caacgttcct gacaacgagc
     1321 ctccttttcg ccaatccatc gacaatcacc gcgagtccct gctcgaacgc tgcgtccgga
     1381 ccggcttcgt cgaaggcgtc tatcgcggcc cgcaacagcg gcgagagcgg agcctgttca
     1441 acggtgccgc cgcgctcgcc ggcatcgctg tcgccggcct gctcctcaag cacggcccca
     1501 acagtgaagt agctgattgt catcagcgca ttgacggcgt ccccggccga aaaacccgcc
     1561 tcgcagagga agcgaagctg cgcgtcggcc gtttccatct gcggtgcgcc cggtcgcgtg
     1621 ccggcatgga tgcgcgcgcc atcgcggtag gcgagcagcg cctgcctgaa gctgcgggca
     1681 ttcccgatca gaaatgagcg ccagtcgtcg tcggctctcg gcaccgaatg cgtatgattc
     1741 tccgccagca tggcttcggc cagtgcgtcg agcagcgccc gcttgttcct gaagtgccag
     1801 taaagcgccg gctgctgaac ccccaaccgt tccgccagtt tgcgtgtcgt cagaccgtct
     1861 acgccgacct cgttcaacag gtccagggcg gcacggatca ctgtattcgg ctgcaacttt
     1921 gtcatgcttg acactttatc actgataaac ataatatgtc caccaactta tcagtgataa
     1981 agaatccgcg cgttcaatcg gaccagcgga ggctggtccg gaggccagac gtgaaaccca
     2041 acatacccct gatcgtaatt ctgagcactg tcgcgctcga cgctgtcggc atcggcctga
     2101 ttatgccggt gctgccgggc ctcctgcgcg atctggttca ctcgaacgac gtcaccgccc
     2161 actatggcat tctgctggcg ctgtatgcgt tggtgcaatt tgcctgcgca cctgtgctgg
     2221 gcgcgctgtc ggatcgtttc gggcggcggc caatcttgct cgtctcgctg gccggcgcca
     2281 gatctgggga accctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc
     2341 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc
     2401 aatatatcct gtcaaacact gatagtttaa actgaaggcg ggaaacgaca atctgatcat
     2461 gagcggagaa ttaagggagt cacgttatga cccccgccga tgacgcggga caagccgttt
     2521 tacgtttgga actgacagaa ccgcaacgtt gaaggagcca ctcagccgcg ggtttctgga
     2581 gtttaatgag ctaagcacat acgtcagaaa ccattattgc gcgttcaaaa gtcgcctaag
     2641 gtcactatca gctagacgga cgcggccgcg ggcccaatat aacaacgacg tcgtaacaga
     2701 taaagcgaag cttgaaggtg catgtgactc cgtcaagatt acgaaaccgc caactaccac
     2761 gcaaattgca attctcaatt tcctagaagg actctccgaa aatgcatcca ataccaaata
     2821 ttacccgtgt cataggcacc aagtgacacc atacatgaac acgcgtcaca atatgactgg
     2881 agaagggttc cacaccttat gctataaaac gccccacacc cctcctcctt ccttcgcagt
     2941 tcaattccaa tatattccat tctctctgtg tatttcccta cctctccctt caaggttagt
     3001 cgatttcttc tgtttttctt cttcgttctt tccatgaatt gtgtatgttc tttgatcaat
     3061 acgatgttga tttgattgtg ttttgtttgg tttcatcgat cttcaatttt cataatcaga
     3121 ttcagctttt attatcttta caacaacgtc cttaatttga tgattcttta atcgtagatt
     3181 tgctctaatt agagcttttt catgtcagat ccctttacaa caagccttaa ttgttgattc
     3241 attaatcgta gattagggct tttttcattg attacttcag atccgttaaa cgtaaccata
     3301 gatcagggct ttttcatgaa ttacttcaga tccgttaaac aacagcctta ttttttatac
     3361 ttctgtggtt tttcaagaaa ttgttcagat ccgttgacaa aaagccttat tcgttgattc
     3421 tatatcgttt ttcgagagat attgctcaga tctgttagca actgccttgt ttgttgattc
     3481 tattgccgtg gattagggtt ttttttcacg agattgcttc agatccgtac ttaagattac
     3541 gtaatggatt ttgattctga tttatctgtg attgttgact cgacaggtac cttcaaacct
     3601 cgaggcgcgc caagctatca aacaagtttg tacaaaaaag caggctccga attcgccctt
     3661 caccatggat tacaaggatg atgatgataa ggattacaag gatgatgatg ataagatggc
     3721 tccaaagaag aagagaaagg ttggaatcca cggagttcca gctgctgata agaagtactc
     3781 tatcggactt gacatcggaa ccaactctgt tggatgggct gttatcaccg atgagtacaa
     3841 ggttccatct aagaagttca aggttcttgg aaacaccgat agacactcta tcaagaagaa
     3901 ccttatcggt gctcttcttt tcgattctgg agagaccgct gaggctacca gattgaagag
     3961 aaccgctaga agaagataca ccagaagaaa gaacagaatc tgctaccttc aggaaatctt
     4021 ctctaacgag atggctaagg ttgatgattc tttcttccac agacttgagg agtctttcct
     4081 tgttgaggag gataagaagc acgagagaca cccaatcttc ggaaacatcg ttgatgaggt
     4141 tgcttaccac gagaagtacc caaccatcta ccaccttaga aagaagttgg ttgattctac
     4201 cgataaggct gatcttagac ttatctacct tgctcttgct cacatgatca agttcagagg
     4261 acacttcctt atcgagggag accttaaccc agataactct gatgttgata agttgttcat
     4321 ccagcttgtt cagacctaca accagctttt cgaggagaac ccaatcaacg cttctggagt
     4381 tgatgctaag gctatccttt ctgctagact ttctaagtct cgtagacttg agaaccttat
     4441 cgctcagctt ccaggagaga agaagaacgg acttttcgga aaccttatcg ctctttctct
     4501 tggacttacc ccaaacttca agtctaactt cgatcttgct gaggatgcta agttgcagct
     4561 ttctaaggat acctacgatg atgatcttga taaccttctt gctcagatcg gagatcagta
     4621 cgctgatctt ttccttgctg ctaagaacct ttctgatgct atccttcttt ctgacatcct
     4681 tagagttaac accgagatca ccaaggctcc actttctgct tctatgatca agagatacga
     4741 tgagcaccac caggatctta cccttttgaa ggctcttgtt agacagcagc ttccagagaa
     4801 gtacaaggaa atcttcttcg atcagtctaa gaacggatac gctggataca tcgatggagg
     4861 agcttctcag gaggagttct acaagttcat caagccaatc cttgagaaga tggatggaac
     4921 cgaggagctt cttgttaagt tgaacagaga ggatcttctt agaaagcaga gaaccttcga
     4981 taacggatct atcccacacc agatccacct tggagagctt cacgctatcc ttcgtagaca
     5041 ggaggatttc tacccattct tgaaggataa cagagagaag atcgagaaga tccttacctt
     5101 cagaatccca tactacgttg gaccacttgc tagaggaaac tctcgtttcg cttggatgac
     5161 cagaaagtct gaggagacca tcaccccttg gaacttcgag gaggtaagtt tctgcttcta
     5221 cctttgatat atatataata attatcatta attagtagta atataatatt tcaaatattt
     5281 ttttcaaaat aaaagaatgt agtatatagc aattgctttt ctgtagttta taagtgtgta
     5341 tattttaatt tataactttt ctaatatatg accaaaattt gttgatgtgc aggttgttga
     5401 taagggagct tctgctcagt ctttcatcga gagaatgacc aacttcgata agaaccttcc
     5461 aaacgagaag gttcttccaa agcactctct tctttacgag tacttcaccg tttacaacga
     5521 gcttaccaag gttaagtacg ttaccgaggg aatgagaaag ccagctttcc tttctggaga
     5581 gcagaagaag gctatcgttg atcttctttt caagaccaac agaaaggtta ccgttaagca
     5641 gttgaaggag gattacttca agaagatcga gtgcttcgat tctgttgaaa tctctggagt
     5701 tgaggataga ttcaacgctt ctcttggaac ctaccacgat cttttgaaga tcatcaagga
     5761 taaggatttc cttgataacg aggagaacga ggacatcctt gaggacatcg ttcttaccct
     5821 tacccttttc gaggatagag agatgatcga ggagagactc aagacctacg ctcacctttt
     5881 cgatgataag gttatgaagc agttgaagag aagaagatac accggatggg gtagactttc
     5941 tcgtaagttg atcaacggaa tcagagataa gcagtctgga aagaccatcc ttgatttctt
     6001 gaagtctgat ggattcgcta acagaaactt catgcagctt atccacgatg attctcttac
     6061 cttcaaggag gacatccaga aggctcaggt ttctggacag ggagattctc ttcacgagca
     6121 catcgctaac cttgctggat ctccagctat caagaaggga atccttcaga ccgttaaggt
     6181 tgttgatgag cttgttaagg ttatgggtag acacaagcca gagaacatcg ttatcgagat
     6241 ggctagagag aaccagacca cccagaaggg acagaagaac tctcgtgaga gaatgaagag
     6301 aatcgaggag ggaatcaagg agcttggatc tcaaatcttg aaggagcacc cagttgagaa
     6361 cacccagctt cagaacgaga agttgtacct ttactacctt cagaacggaa gagatatgta
     6421 cgttgatcag gagcttgaca tcaacagact ttctgattac gatgttgatc acatcgttcc
     6481 acagtctttc ttgaaggatg attctatcga taacaaggtt cttacccgtt ctgataagaa
     6541 cagaggaaag tctgataacg ttccatctga ggaggttgtt aagaagatga agaactactg
     6601 gagacagctt cttaacgcta agttgatcac ccagagaaag ttcgataacc ttaccaaggc
     6661 tgagagagga ggactttctg agcttgataa ggctggattc atcaagagac agcttgttga
     6721 gaccagacag atcaccaagc acgttgctca gatccttgat tctcgtatga acaccaagta
     6781 cgatgagaac gataagttga tcagagaggt taaggttatc accttgaagt ctaagttggt
     6841 ttctgatttc agaaaggatt tccagttcta caaggttaga gagatcaaca actaccacca
     6901 cgctcacgat gcttacctta acgctgttgt tggaaccgct cttatcaaga agtacccaaa
     6961 gttggagtct gagttcgttt acggagatta caaggtttac gatgttagaa agatgatcgc
     7021 taagtctgag caggagatcg gaaaggctac cgctaagtac ttcttctact ctaacatcat
     7081 gaacttcttc aagaccgaga tcacccttgc taacggagag atcagaaaga gaccacttat
     7141 cgagaccaac ggagagaccg gagagatcgt ttgggataag ggaagagatt tcgctaccgt
     7201 tagaaaggtt ctttctatgc cacaggttaa catcgttaag aaaaccgagg ttcagaccgg
     7261 aggattctct aaggagtcta tccttccaaa gagaaactct gataagttga tcgctagaaa
     7321 gaaggattgg gacccaaaga agtacggagg attcgattct ccaaccgttg cttactctgt
     7381 tcttgttgtt gctaaggttg agaagggaaa gtctaagaag ttgaagtctg ttaaggagct
     7441 tcttggaatc accatcatgg agcgttcttc tttcgagaag aacccaatcg atttccttga
     7501 ggctaaggga tacaaggagg ttaagaagga tcttatcatc aagttgccaa agtactctct
     7561 tttcgagctt gagaacggaa gaaagagaat gcttgcttct gctggagagc ttcagaaggg
     7621 aaacgagctt gctcttccat ctaagtacgt taacttcctt taccttgctt ctcactacga
     7681 gaagttgaag ggatctccag aggataacga gcagaagcag cttttcgttg agcagcacaa
     7741 gcactacctt gatgagatca tcgagcaaat ctctgagttc tctaagagag ttatccttgc
     7801 tgatgctaac cttgataagg ttctttctgc ttacaacaag cacagagata agccaatcag
     7861 agagcaggct gagaacatca tccacctttt cacccttacc aaccttggtg ctccagctgc
     7921 tttcaagtac ttcgatacca ccatcgatag aaaaagatac acctctacca aggaggttct
     7981 tgatgctacc cttatccacc agtctatcac cggactttac gagaccagaa tcgatctttc
     8041 tcagcttgga ggagataaga gaccagctgc taccaagaag gctggacagg ctaagaagaa
     8101 gaagtgagac gtccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg
     8161 ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa
     8221 ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat
     8281 tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc
     8341 gcgcggtgtc atctatgtta ctagatcggg aattgatccc ccctcgacag cttccggaaa
     8401 gggcgaattc gcaactttgt atacaaaagt tgccgagctc gcggccgcat gttgttgtta
     8461 ccagaaagta aataaatgtt caatctctga tgttctcaag taagtgagtt ttattgggaa
     8521 taatattaac ttatgttctt cttgcatttg atttctttgc cgctctcttc ttctatctta
     8581 aatctgtgta tactatttca ctattgggct ttttattagt ctataatggg actcaaaata
     8641 aggctttggc ccacatcaaa aagataagtc acaaatcaaa actaaattca gagtcttttc
     8701 tcccacatcg gtcactgtac tcattttgtg tttgtttata tattacacga accgatcttt
     8761 ggtacggatt tctcagctag atgccagttt tagagctaga aatagcaagt taaaataagg
     8821 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc tagacccagc
     8881 tttcttgtac aaagttggca ttaccatggc ttcgttgaac aacggaaact cgacttgcct
     8941 tccgcacaat acatcatttc ttcttagctt tttttcttct tcttcgttca tacagttttt
     9001 ttttgtttat cagcttacat tttcttgaac cgtagctttc gttttcttct ttttaacttt
     9061 ccattcggag tttttgtatc ttgtttcata gtttgtccca ggattagaat gattaggcat
     9121 cgaaccttca agaatttgat tgaataaaac atcttcattc ttaagatatg aagataatct
     9181 tcaaaaggcc cctgggaatc tgaaagaaga gaagcaggcc catttatatg ggaaagaaca
     9241 atagtatttc ttatataggc ccatttaagt tgaaaacaat cttcaaaagt cccacatcgc
     9301 ttagataaga aaacgaagct gagtttatat acagctagag tcgaagtagt gattgggact
     9361 tctcaaagca tgttgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     9421 caacttgaaa aagtggcacc gagtcggtgc tttttttcta gacccagctt tcttgtacaa
     9481 agttggcatt actgcaggtc gacgacccag ctttcttgta caaagtggtt cgataattct
     9541 taattaacta gttctaatct agagtcctgc tttaatgaga tatgcgagac gcctatgatc
     9601 gcatgatatt tgctttcaat tctgttgtgc acgttgtaaa aaacctgagc atgtgtagct
     9661 cagatcctta ccgccggttt cggttcattc taatgaatat atcacccgtt actatcgtat
     9721 ttttatgaat aatattctcc gttcaattta ctgattgtac cctactactt atatgtacaa
     9781 tattaaaatg aaaacaatat attgtgctga ataggtttat agcgacatct atgatagagc
     9841 gccacaataa caaacaattg cgttttatta ttacaaatcc aattttaaaa aaagcggcag
     9901 aaccggtcaa acctaaaaga ctgattacat aaatcttatt caaatttcaa aagtgcccca
     9961 ggggctagta tctacgacac accgagcggc gaactaataa cgctcactga agggaactcc
    10021 ggttccccgc cggcgcgcat gggtgagatt ccttgaagtt gagtattggc cgtccgctct
    10081 accgaaagtt acgggcacca ttcaacccgg tccagcacgg cggccgggta accgacttgc
    10141 tgccccgaga attatgcagc atttttttgg tgtatgtggg ccccaaatga agtgcaggtc
    10201 aaaccttgac agtgacgaca aatcgttggg cgggtccagg gcgaattttg cgacaacatg
    10261 tcgaggctca gcaggacctg caggcatgca agctagtgaa gaatcgattt gctacgtatt
    10321 gcccctgcag gtccccagat tagccttttc aatttcagaa agaatgctaa cccacagatg
    10381 gttagagagg cttacgcagc aggtctcatc aagacgatct acccgagcaa taatctccag
    10441 gaaatcaaat accttcccaa gaaggttaaa gatgcagtca aaagattcag gactaactgc
    10501 atcaagaaca cagagaaaga tatatttctc aagatcagaa gtactattcc agtatggacg
    10561 attcaaggct tgcttcacaa accaaggcaa gtaatagaga ttggagtctc taaaaaggta
    10621 gttcccactg aatcaaaggc catggagtca aagattcaaa tagaggacct aacagaactc
    10681 gccgtaaaga ctggcgaaca gttcatacag agtctcttac gactcaatga caagaagaaa
    10741 atcttcgtca acatggtgga gcacgacaca cttgtctact ccaaaaatat caaagataca
    10801 gtctcagaag accaaagggc aattgagact tttcaacaaa gggtaatatc cggaaacctc
    10861 ctcggattcc attgcccagc tatctgtcac tttattgtga agatagtgga aaaggaaggt
    10921 ggctcctaca aatgccatca ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc
    10981 gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt
    11041 ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt aagggatgac
    11101 gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg
    11161 gagagaacac gggggactct agaggatcca tgagcccaga acgacgcccg gccgacatcc
    11221 gccgtgccac cgaggcggac atgccggcgg tctgcaccat cgtcaaccac tacatcgaga
    11281 caagcacggt caacttccgt accgagccgc aggaaccgca ggagtggacg gacgacctcg
    11341 tccgtctgcg ggagcgctat ccctggctcg tcgccgaggt ggacggcgag gtcgccggca
    11401 tcgcctacgc gggcccctgg aaggcacgca acgcctacga ctggacggcc gagtcgaccg
    11461 tgtacgtctc cccccgccac cagcggacgg gactgggctc cacgctctac acccacctgc
    11521 tgaagtccct ggaggcacag ggcttcaaga gcgtggtcgc tgtcatcggg ctgcccaacg
    11581 acccgagcgt gcgcatgcac gaggcgctcg gatatgcccc ccgcggcatg ctgcgggcgg
    11641 ccggcttcaa gcacgggaac tggcatgacg tgggtttctg gcagctggac ttcagcctgc
    11701 cggtaccgcc ccgtccggtc ctgcccgtca ccgagatctg aacggagtgc gcgtggcatc
    11761 gcccgagttg gagctggtac gggaactcat cgaactcaac tggcataccc gcaatggtga
    11821 ggtggaaccg cggcggatcg cgtacgaccg tgcccaggag ggggtaccga gctcgaattt
    11881 ccccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
    11941 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
    12001 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
    12061 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
    12121 atctatgtta ctagatcggg aattaattca ctggccgtcg ttttacaacg tcgtgactgg
    12181 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
    12241 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
    12301 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
    12361 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
    12421 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
    12481 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
    12541 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcgga
    12601 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa
    12661 ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga
    12721 aaaaccaccc cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa
    12781 tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct
    12841 cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag
    12901 agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac
    12961 taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga
    13021 tgacagagcg ttgctgcctg tgatcaaata tcatctccct cgcagagatc cgaattatca
    13081 gccttcttat tcatttctcg cttaaccgtg acaggctgtc gatcttgaga actatgccga
    13141 cataatagga aatcgctgga taaagccgct gaggaagctg agtggcgcta tttctttaga
    13201 agtgaacgtt gacgatatca actcccctat ccattgctca ccgaatggta caggtcgggg
    13261 acccgaagtt ccgactgtcg gcctgatgca tccccggctg atcgacccca gatctggggc
    13321 tgagaaagcc cagtaaggaa acaactgtag gttcgagtcg cgagatcccc cggaaccaaa
    13381 ggaagtaggt taaacccgct ccgatcaggc cgagccacgc caggccgaga acattggttc
    13441 ctgtaggcat cgggattggc ggatcaaaca ctaaagctac tggaacgagc agaagtcctc
    13501 cggccgccag ttgccaggcg gtaaaggtga gcagaggcac gggaggttgc cacttgcggg
    13561 tcagcacggt tccgaacgcc atggaaaccg cccccgccag gcccgctgcg acgccgacag
    13621 gatctagcgc tgcgtttggt gtcaacacca acagcgccac gcccgcagtt ccgcaaatag
    13681 cccccaggac cgccatcaat cgtatcgggc tacctagcag agcggcagag atgaacacga
    13741 ccatcagcgg ctgcacagcg cctaccgtcg ccgcgacccc gcccggcagg cggtagaccg
    13801 aaataaacaa caagctccag aatagcgaaa tattaagtgc gccgaggatg aagatgcgca
    13861 tccaccagat tcccgttgga atctgtcgga cgatcatcac gagcaataaa cccgccggca
    13921 acgcccgcag cagcataccg gcgacccctc ggcctcgctg ttcgggctcc acgaaaacgc
    13981 cggacagatg cgccttgtga gcgtccttgg ggccgtcctc ctgtttgaag accgacagcc
    14041 caatgatctc gccgtcgatg taggcgccga atgccacggc atctcgcaac cgttcagcga
    14101 acgcctccat gggctttttc tcctcgtgct cgtaaacgga cccgaacatc tctggagctt
    14161 tcttcagggc cgacaatcgg atctcgcgga aatcctgcac gtcggccgct ccaagccgtc
    14221 gaatctgagc cttaatcaca attgtcaatt ttaatcctct gtttatcggc agttcgtaga
    14281 gcgcgccgtg cgtcccgagc gatactgagc gaagcaagtg cgtcgagcag tgcccgcttg
    14341 ttcctgaaat gccagtaaag cgctggctgc tgaaccccca gccggaactg accccacaag
    14401 gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct
    14461 gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga
    14521 atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga
    14581 gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat
    14641 gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg
    14701 gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga
    14761 ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag
    14821 gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag
    14881 ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac
    14941 ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt
    15001 cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt
    15061 gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc
    15121 cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg
    15181 tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt
    15241 ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc
    15301 tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc
    15361 agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga
    15421 gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc
    15481 ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa
    15541 cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc
    15601 ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat
    15661 gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc
    15721 ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg cctgagagcc
    15781 aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca
    15841 catctaggta ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag
    15901 gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc
    15961 tgatcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa
    16021 gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc
    16081 cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc
    16141 gcggatcttt aaatggagtg tcttcttccc agttttcgca atccacatcg gccagatcgt
    16201 tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat
    16261 ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt
    16321 cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga
    16381 gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc
    16441 cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc
    16501 ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta tataccttag
    16561 caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt
    16621 ccggtgatat tctcatttta gccatttatt atttccttcc tcttttctac agtatttaaa
    16681 gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta
    16741 aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat
    16801 agtatcgacg gagccgattt tgaaaccaca attatgggtg atgctgccaa cttactgatt
    16861 tagtgtatga tggtgttttt gaggtgctcc agtggcttct gtttctatca gctgtccctc
    16921 ctgttcagct actgacgggg tggtgcgtaa cggcaaaagc accgccggac atcagcgcta
    16981 tctctgctct cactgccgta aaacatggca actgcagttc acttacaccg cttctcaacc
    17041 cggtacgcac cagaaaatca ttgatatggc catgaatggc gttggatgcc gggcaacagc
    17101 ccgcattatg ggcgttggcc tcaacacgat tttacgtcac ttaaaaaact caggccgcag
    17161 tcggtaacct cgcgcataca gccgggcagt gacgtcatcg tctgcgcgga aatggacgaa
    17221 cagtggggct atgtcggggc taaatcgcgc cagcgctggc tgttttacgc gtatgacagt
    17281 ctccggaaga cggttgttgc gcacgtattc ggtgaacgca ctatggcgac gctggggcgt
    17341 cttatgagcc tgctgtcacc ctttgacgtg gtgatatgga tgacggatgg ctggccgctg
    17401 tatgaatccc gcctgaaggg aaagctgcac gtaatcagca agcgatatac gcagcgaatt
    17461 gagcggcata acctgaatct gaggcagcac ctggcacggc tgggacggaa gtcgctgtcg
    17521 ttctcaaaat cggtggagct gcatgacaaa gtcatcgggc attatctgaa cataaaacac
    17581 tatcaataag ttggagtcat tacccaatta tgatagaatt tacaagctat aaggttattg
    17641 tcctgggttt caagcattag tccatgcaag tttttatgct ttgcccattc tatagatata
    17701 ttgataagcg cgctgcctat gccttgcccc ctgaaatcct tacatacggc gatatcttct
    17761 atataaaaga tatattatct tatcagtatt gtcaatatat tcaaggcaat ctgcctcctc
    17821 atcctcttca tcctcttcgt cttggtagct ttttaaatat ggcgcttcat agagtaattc
    17881 tgtaaaggtc caattctcgt tttcatacct cggtataatc ttacctatca cctcaaatgg
    17941 ttcgctgggt ttatcgcacc cccgaacacg agcacggcac ccgcgaccac tatgccaaga
    18001 atgcccaagg taaaaattgc cggccccgcc atgaagtccg tgaatgcccc gacggccgaa
    18061 gtgaagggca ggccgccacc caggccgccg ccctcactgc ccggcacctg gtcgctgaat
    18121 gtcgatgcca gcacctgcgg cacgtcaatg cttccgggcg tcgcgctcgg gctgatcgcc
    18181 catcccgtta ctgccccgat cccggcaatg gcaaggactg ccagcgctgc catttttggg
    18241 gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg
    18301 ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag
    18361 ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac
    18421 aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt
    18481 ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag
    18541 tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag
    18601 ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt
    18661 ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc
    18721 atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct
    18781 gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgg ccgcggtgtc
    18841 tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg gccatagacg gccgccagcc
    18901 cagcggcgag ggcaaccagc ccggtgagcg tcggaaaggc gctcggtctt gccttgctcg
    18961 tcggtgatgt acttcaccag ctccgcga
//
