LOCUS       sequence-202692-       18989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g020630 (FmoI). Also 
            expresses Cas9 from Gmubi promoter from Gmubi promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18989)
  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 18989)
  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: pSC19
FEATURES             Location/Qualifiers
     source          1..18989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(111..482)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            515..624
                     /note="incP origin of transfer"
     primer_bind     847..866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1100..1117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1305..1955)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2420..2444
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3652..3676
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3698..3745
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3752..3772
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3797..5233,5423..8086)
                     /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          5234..5422
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8087..8134
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8145..8397
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8442..8462
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8817..8892
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8904..8928)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9406..9481
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9493..9517)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9536..9560)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9598..10305
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10855..11200
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11143..11163
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11221..11772
                     /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      11916..12168
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12181..12198)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12181..12197)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12190..12212)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12337..12356)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12547..12568
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12815..12839
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14435..15583)
                     /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(15882..16676)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16868..17635
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18258..18973
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cgtcggtgat gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga
       61 cgtgcttggc aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct
      121 gccctcgggc ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc
      181 ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg
      241 agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg
      301 cggacgtgct catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg
      361 taggccgaca ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct
      421 ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc
      481 atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc
      541 ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt
      601 gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca
      661 cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc
      721 gctataatga ccccgaagca gggttatgca gcggaaaagc gccacgcttc ccgaagggag
      781 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
      841 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
      901 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      961 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     1021 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     1081 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gccagaaggc
     1141 cgccagagag gccgagcgcg gccgtgaggc ttggacgcta gggcagggca tgaaaaagcc
     1201 cgtagcgggc tgctacgggc gtctgacgcg gtggaaaggg ggaggggatg ttgtctacat
     1261 ggctctgctg tagtgagtgg gttgcgctcc ggcagcggtc ctgatcaatc gtcacccttt
     1321 ctcggtcctt caacgttcct gacaacgagc ctccttttcg ccaatccatc gacaatcacc
     1381 gcgagtccct gctcgaacgc tgcgtccgga ccggcttcgt cgaaggcgtc tatcgcggcc
     1441 cgcaacagcg gcgagagcgg agcctgttca acggtgccgc cgcgctcgcc ggcatcgctg
     1501 tcgccggcct gctcctcaag cacggcccca acagtgaagt agctgattgt catcagcgca
     1561 ttgacggcgt ccccggccga aaaacccgcc tcgcagagga agcgaagctg cgcgtcggcc
     1621 gtttccatct gcggtgcgcc cggtcgcgtg ccggcatgga tgcgcgcgcc atcgcggtag
     1681 gcgagcagcg cctgcctgaa gctgcgggca ttcccgatca gaaatgagcg ccagtcgtcg
     1741 tcggctctcg gcaccgaatg cgtatgattc tccgccagca tggcttcggc cagtgcgtcg
     1801 agcagcgccc gcttgttcct gaagtgccag taaagcgccg gctgctgaac ccccaaccgt
     1861 tccgccagtt tgcgtgtcgt cagaccgtct acgccgacct cgttcaacag gtccagggcg
     1921 gcacggatca ctgtattcgg ctgcaacttt gtcatgcttg acactttatc actgataaac
     1981 ataatatgtc caccaactta tcagtgataa agaatccgcg cgttcaatcg gaccagcgga
     2041 ggctggtccg gaggccagac gtgaaaccca acatacccct gatcgtaatt ctgagcactg
     2101 tcgcgctcga cgctgtcggc atcggcctga ttatgccggt gctgccgggc ctcctgcgcg
     2161 atctggttca ctcgaacgac gtcaccgccc actatggcat tctgctggcg ctgtatgcgt
     2221 tggtgcaatt tgcctgcgca cctgtgctgg gcgcgctgtc ggatcgtttc gggcggcggc
     2281 caatcttgct cgtctcgctg gccggcgcca gatctgggga accctgtggt tggcatgcac
     2341 atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga ttattctaat
     2401 aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcaaacact gatagtttaa
     2461 actgaaggcg ggaaacgaca atctgatcat gagcggagaa ttaagggagt cacgttatga
     2521 cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa ccgcaacgtt
     2581 gaaggagcca ctcagccgcg ggtttctgga gtttaatgag ctaagcacat acgtcagaaa
     2641 ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagacgga cgcggccgcg
     2701 ggcccaatat aacaacgacg tcgtaacaga taaagcgaag cttgaaggtg catgtgactc
     2761 cgtcaagatt acgaaaccgc caactaccac gcaaattgca attctcaatt tcctagaagg
     2821 actctccgaa aatgcatcca ataccaaata ttacccgtgt cataggcacc aagtgacacc
     2881 atacatgaac acgcgtcaca atatgactgg agaagggttc cacaccttat gctataaaac
     2941 gccccacacc cctcctcctt ccttcgcagt tcaattccaa tatattccat tctctctgtg
     3001 tatttcccta cctctccctt caaggttagt cgatttcttc tgtttttctt cttcgttctt
     3061 tccatgaatt gtgtatgttc tttgatcaat acgatgttga tttgattgtg ttttgtttgg
     3121 tttcatcgat cttcaatttt cataatcaga ttcagctttt attatcttta caacaacgtc
     3181 cttaatttga tgattcttta atcgtagatt tgctctaatt agagcttttt catgtcagat
     3241 ccctttacaa caagccttaa ttgttgattc attaatcgta gattagggct tttttcattg
     3301 attacttcag atccgttaaa cgtaaccata gatcagggct ttttcatgaa ttacttcaga
     3361 tccgttaaac aacagcctta ttttttatac ttctgtggtt tttcaagaaa ttgttcagat
     3421 ccgttgacaa aaagccttat tcgttgattc tatatcgttt ttcgagagat attgctcaga
     3481 tctgttagca actgccttgt ttgttgattc tattgccgtg gattagggtt ttttttcacg
     3541 agattgcttc agatccgtac ttaagattac gtaatggatt ttgattctga tttatctgtg
     3601 attgttgact cgacaggtac cttcaaacct cgaggcgcgc caagctatca aacaagtttg
     3661 tacaaaaaag caggctccga attcgccctt caccatggat tacaaggatg atgatgataa
     3721 ggattacaag gatgatgatg ataagatggc tccaaagaag aagagaaagg ttggaatcca
     3781 cggagttcca gctgctgata agaagtactc tatcggactt gacatcggaa ccaactctgt
     3841 tggatgggct gttatcaccg atgagtacaa ggttccatct aagaagttca aggttcttgg
     3901 aaacaccgat agacactcta tcaagaagaa ccttatcggt gctcttcttt tcgattctgg
     3961 agagaccgct gaggctacca gattgaagag aaccgctaga agaagataca ccagaagaaa
     4021 gaacagaatc tgctaccttc aggaaatctt ctctaacgag atggctaagg ttgatgattc
     4081 tttcttccac agacttgagg agtctttcct tgttgaggag gataagaagc acgagagaca
     4141 cccaatcttc ggaaacatcg ttgatgaggt tgcttaccac gagaagtacc caaccatcta
     4201 ccaccttaga aagaagttgg ttgattctac cgataaggct gatcttagac ttatctacct
     4261 tgctcttgct cacatgatca agttcagagg acacttcctt atcgagggag accttaaccc
     4321 agataactct gatgttgata agttgttcat ccagcttgtt cagacctaca accagctttt
     4381 cgaggagaac ccaatcaacg cttctggagt tgatgctaag gctatccttt ctgctagact
     4441 ttctaagtct cgtagacttg agaaccttat cgctcagctt ccaggagaga agaagaacgg
     4501 acttttcgga aaccttatcg ctctttctct tggacttacc ccaaacttca agtctaactt
     4561 cgatcttgct gaggatgcta agttgcagct ttctaaggat acctacgatg atgatcttga
     4621 taaccttctt gctcagatcg gagatcagta cgctgatctt ttccttgctg ctaagaacct
     4681 ttctgatgct atccttcttt ctgacatcct tagagttaac accgagatca ccaaggctcc
     4741 actttctgct tctatgatca agagatacga tgagcaccac caggatctta cccttttgaa
     4801 ggctcttgtt agacagcagc ttccagagaa gtacaaggaa atcttcttcg atcagtctaa
     4861 gaacggatac gctggataca tcgatggagg agcttctcag gaggagttct acaagttcat
     4921 caagccaatc cttgagaaga tggatggaac cgaggagctt cttgttaagt tgaacagaga
     4981 ggatcttctt agaaagcaga gaaccttcga taacggatct atcccacacc agatccacct
     5041 tggagagctt cacgctatcc ttcgtagaca ggaggatttc tacccattct tgaaggataa
     5101 cagagagaag atcgagaaga tccttacctt cagaatccca tactacgttg gaccacttgc
     5161 tagaggaaac tctcgtttcg cttggatgac cagaaagtct gaggagacca tcaccccttg
     5221 gaacttcgag gaggtaagtt tctgcttcta cctttgatat atatataata attatcatta
     5281 attagtagta atataatatt tcaaatattt ttttcaaaat aaaagaatgt agtatatagc
     5341 aattgctttt ctgtagttta taagtgtgta tattttaatt tataactttt ctaatatatg
     5401 accaaaattt gttgatgtgc aggttgttga taagggagct tctgctcagt ctttcatcga
     5461 gagaatgacc aacttcgata agaaccttcc aaacgagaag gttcttccaa agcactctct
     5521 tctttacgag tacttcaccg tttacaacga gcttaccaag gttaagtacg ttaccgaggg
     5581 aatgagaaag ccagctttcc tttctggaga gcagaagaag gctatcgttg atcttctttt
     5641 caagaccaac agaaaggtta ccgttaagca gttgaaggag gattacttca agaagatcga
     5701 gtgcttcgat tctgttgaaa tctctggagt tgaggataga ttcaacgctt ctcttggaac
     5761 ctaccacgat cttttgaaga tcatcaagga taaggatttc cttgataacg aggagaacga
     5821 ggacatcctt gaggacatcg ttcttaccct tacccttttc gaggatagag agatgatcga
     5881 ggagagactc aagacctacg ctcacctttt cgatgataag gttatgaagc agttgaagag
     5941 aagaagatac accggatggg gtagactttc tcgtaagttg atcaacggaa tcagagataa
     6001 gcagtctgga aagaccatcc ttgatttctt gaagtctgat ggattcgcta acagaaactt
     6061 catgcagctt atccacgatg attctcttac cttcaaggag gacatccaga aggctcaggt
     6121 ttctggacag ggagattctc ttcacgagca catcgctaac cttgctggat ctccagctat
     6181 caagaaggga atccttcaga ccgttaaggt tgttgatgag cttgttaagg ttatgggtag
     6241 acacaagcca gagaacatcg ttatcgagat ggctagagag aaccagacca cccagaaggg
     6301 acagaagaac tctcgtgaga gaatgaagag aatcgaggag ggaatcaagg agcttggatc
     6361 tcaaatcttg aaggagcacc cagttgagaa cacccagctt cagaacgaga agttgtacct
     6421 ttactacctt cagaacggaa gagatatgta cgttgatcag gagcttgaca tcaacagact
     6481 ttctgattac gatgttgatc acatcgttcc acagtctttc ttgaaggatg attctatcga
     6541 taacaaggtt cttacccgtt ctgataagaa cagaggaaag tctgataacg ttccatctga
     6601 ggaggttgtt aagaagatga agaactactg gagacagctt cttaacgcta agttgatcac
     6661 ccagagaaag ttcgataacc ttaccaaggc tgagagagga ggactttctg agcttgataa
     6721 ggctggattc atcaagagac agcttgttga gaccagacag atcaccaagc acgttgctca
     6781 gatccttgat tctcgtatga acaccaagta cgatgagaac gataagttga tcagagaggt
     6841 taaggttatc accttgaagt ctaagttggt ttctgatttc agaaaggatt tccagttcta
     6901 caaggttaga gagatcaaca actaccacca cgctcacgat gcttacctta acgctgttgt
     6961 tggaaccgct cttatcaaga agtacccaaa gttggagtct gagttcgttt acggagatta
     7021 caaggtttac gatgttagaa agatgatcgc taagtctgag caggagatcg gaaaggctac
     7081 cgctaagtac ttcttctact ctaacatcat gaacttcttc aagaccgaga tcacccttgc
     7141 taacggagag atcagaaaga gaccacttat cgagaccaac ggagagaccg gagagatcgt
     7201 ttgggataag ggaagagatt tcgctaccgt tagaaaggtt ctttctatgc cacaggttaa
     7261 catcgttaag aaaaccgagg ttcagaccgg aggattctct aaggagtcta tccttccaaa
     7321 gagaaactct gataagttga tcgctagaaa gaaggattgg gacccaaaga agtacggagg
     7381 attcgattct ccaaccgttg cttactctgt tcttgttgtt gctaaggttg agaagggaaa
     7441 gtctaagaag ttgaagtctg ttaaggagct tcttggaatc accatcatgg agcgttcttc
     7501 tttcgagaag aacccaatcg atttccttga ggctaaggga tacaaggagg ttaagaagga
     7561 tcttatcatc aagttgccaa agtactctct tttcgagctt gagaacggaa gaaagagaat
     7621 gcttgcttct gctggagagc ttcagaaggg aaacgagctt gctcttccat ctaagtacgt
     7681 taacttcctt taccttgctt ctcactacga gaagttgaag ggatctccag aggataacga
     7741 gcagaagcag cttttcgttg agcagcacaa gcactacctt gatgagatca tcgagcaaat
     7801 ctctgagttc tctaagagag ttatccttgc tgatgctaac cttgataagg ttctttctgc
     7861 ttacaacaag cacagagata agccaatcag agagcaggct gagaacatca tccacctttt
     7921 cacccttacc aaccttggtg ctccagctgc tttcaagtac ttcgatacca ccatcgatag
     7981 aaaaagatac acctctacca aggaggttct tgatgctacc cttatccacc agtctatcac
     8041 cggactttac gagaccagaa tcgatctttc tcagcttgga ggagataaga gaccagctgc
     8101 taccaagaag gctggacagg ctaagaagaa gaagtgagac gtccgatcgt tcaaacattt
     8161 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
     8221 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
     8281 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
     8341 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg
     8401 aattgatccc ccctcgacag cttccggaaa gggcgaattc gcaactttgt atacaaaagt
     8461 tgccgagctc gcggccgcat gttgttgtta ccagaaagta aataaatgtt caatctctga
     8521 tgttctcaag taagtgagtt ttattgggaa taatattaac ttatgttctt cttgcatttg
     8581 atttctttgc cgctctcttc ttctatctta aatctgtgta tactatttca ctattgggct
     8641 ttttattagt ctataatggg actcaaaata aggctttggc ccacatcaaa aagataagtc
     8701 acaaatcaaa actaaattca gagtcttttc tcccacatcg gtcactgtac tcattttgtg
     8761 tttgtttata tattacacga accgatcttt ggtacggaag ccactgattc aattgggttt
     8821 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     8881 ccgagtcggt gctttttttc tagacccagc tttcttgtac aaagttggca ttaccatggc
     8941 ttcgttgaac aacggaaact cgacttgcct tccgcacaat acatcatttc ttcttagctt
     9001 tttttcttct tcttcgttca tacagttttt ttttgtttat cagcttacat tttcttgaac
     9061 cgtagctttc gttttcttct ttttaacttt ccattcggag tttttgtatc ttgtttcata
     9121 gtttgtccca ggattagaat gattaggcat cgaaccttca agaatttgat tgaataaaac
     9181 atcttcattc ttaagatatg aagataatct tcaaaaggcc cctgggaatc tgaaagaaga
     9241 gaagcaggcc catttatatg ggaaagaaca atagtatttc ttatataggc ccatttaagt
     9301 tgaaaacaat cttcaaaagt cccacatcgc ttagataaga aaacgaagct gagtttatat
     9361 acagctagag tcgaagtagt gattgggatg cttacctcct gaccagtttt agagctagaa
     9421 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     9481 ctttttttct agacccagct ttcttgtaca aagttggcat tactgcaggt cgacgaccca
     9541 gctttcttgt acaaagtggt tcgataattc ttaattaact agttctaatc tagagtcctg
     9601 ctttaatgag atatgcgaga cgcctatgat cgcatgatat ttgctttcaa ttctgttgtg
     9661 cacgttgtaa aaaacctgag catgtgtagc tcagatcctt accgccggtt tcggttcatt
     9721 ctaatgaata tatcacccgt tactatcgta tttttatgaa taatattctc cgttcaattt
     9781 actgattgta ccctactact tatatgtaca atattaaaat gaaaacaata tattgtgctg
     9841 aataggttta tagcgacatc tatgatagag cgccacaata acaaacaatt gcgttttatt
     9901 attacaaatc caattttaaa aaaagcggca gaaccggtca aacctaaaag actgattaca
     9961 taaatcttat tcaaatttca aaagtgcccc aggggctagt atctacgaca caccgagcgg
    10021 cgaactaata acgctcactg aagggaactc cggttccccg ccggcgcgca tgggtgagat
    10081 tccttgaagt tgagtattgg ccgtccgctc taccgaaagt tacgggcacc attcaacccg
    10141 gtccagcacg gcggccgggt aaccgacttg ctgccccgag aattatgcag catttttttg
    10201 gtgtatgtgg gccccaaatg aagtgcaggt caaaccttga cagtgacgac aaatcgttgg
    10261 gcgggtccag ggcgaatttt gcgacaacat gtcgaggctc agcaggacct gcaggcatgc
    10321 aagctagtga agaatcgatt tgctacgtat tgcccctgca ggtccccaga ttagcctttt
    10381 caatttcaga aagaatgcta acccacagat ggttagagag gcttacgcag caggtctcat
    10441 caagacgatc tacccgagca ataatctcca ggaaatcaaa taccttccca agaaggttaa
    10501 agatgcagtc aaaagattca ggactaactg catcaagaac acagagaaag atatatttct
    10561 caagatcaga agtactattc cagtatggac gattcaaggc ttgcttcaca aaccaaggca
    10621 agtaatagag attggagtct ctaaaaaggt agttcccact gaatcaaagg ccatggagtc
    10681 aaagattcaa atagaggacc taacagaact cgccgtaaag actggcgaac agttcataca
    10741 gagtctctta cgactcaatg acaagaagaa aatcttcgtc aacatggtgg agcacgacac
    10801 acttgtctac tccaaaaata tcaaagatac agtctcagaa gaccaaaggg caattgagac
    10861 ttttcaacaa agggtaatat ccggaaacct cctcggattc cattgcccag ctatctgtca
    10921 ctttattgtg aagatagtgg aaaaggaagg tggctcctac aaatgccatc attgcgataa
    10981 aggaaaggcc atcgttgaag atgcctctgc cgacagtggt cccaaagatg gacccccacc
    11041 cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
    11101 atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc
    11161 ttcctctata taaggaagtt catttcattt ggagagaaca cgggggactc tagaggatcc
    11221 atgagcccag aacgacgccc ggccgacatc cgccgtgcca ccgaggcgga catgccggcg
    11281 gtctgcacca tcgtcaacca ctacatcgag acaagcacgg tcaacttccg taccgagccg
    11341 caggaaccgc aggagtggac ggacgacctc gtccgtctgc gggagcgcta tccctggctc
    11401 gtcgccgagg tggacggcga ggtcgccggc atcgcctacg cgggcccctg gaaggcacgc
    11461 aacgcctacg actggacggc cgagtcgacc gtgtacgtct ccccccgcca ccagcggacg
    11521 ggactgggct ccacgctcta cacccacctg ctgaagtccc tggaggcaca gggcttcaag
    11581 agcgtggtcg ctgtcatcgg gctgcccaac gacccgagcg tgcgcatgca cgaggcgctc
    11641 ggatatgccc cccgcggcat gctgcgggcg gccggcttca agcacgggaa ctggcatgac
    11701 gtgggtttct ggcagctgga cttcagcctg ccggtaccgc cccgtccggt cctgcccgtc
    11761 accgagatct gaacggagtg cgcgtggcat cgcccgagtt ggagctggta cgggaactca
    11821 tcgaactcaa ctggcatacc cgcaatggtg aggtggaacc gcggcggatc gcgtacgacc
    11881 gtgcccagga gggggtaccg agctcgaatt tccccgatcg ttcaaacatt tggcaataaa
    11941 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
    12001 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
    12061 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
    12121 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattaattc
    12181 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
    12241 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
    12301 cccttcccaa cagttgcgca gcctgaatgg cgcccgctcc tttcgctttc ttcccttcct
    12361 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
    12421 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
    12481 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    12541 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
    12601 ttgatttata agggattttg ccgatttcgg aaccaccatc aaacaggatt ttcgcctgct
    12661 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
    12721 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ccagtacatt aaaaacgtcc
    12781 gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac
    12841 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
    12901 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
    12961 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
    13021 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaaat
    13081 atcatctccc tcgcagagat ccgaattatc agccttctta ttcatttctc gcttaaccgt
    13141 gacaggctgt cgatcttgag aactatgccg acataatagg aaatcgctgg ataaagccgc
    13201 tgaggaagct gagtggcgct atttctttag aagtgaacgt tgacgatatc aactccccta
    13261 tccattgctc accgaatggt acaggtcggg gacccgaagt tccgactgtc ggcctgatgc
    13321 atccccggct gatcgacccc agatctgggg ctgagaaagc ccagtaagga aacaactgta
    13381 ggttcgagtc gcgagatccc ccggaaccaa aggaagtagg ttaaacccgc tccgatcagg
    13441 ccgagccacg ccaggccgag aacattggtt cctgtaggca tcgggattgg cggatcaaac
    13501 actaaagcta ctggaacgag cagaagtcct ccggccgcca gttgccaggc ggtaaaggtg
    13561 agcagaggca cgggaggttg ccacttgcgg gtcagcacgg ttccgaacgc catggaaacc
    13621 gcccccgcca ggcccgctgc gacgccgaca ggatctagcg ctgcgtttgg tgtcaacacc
    13681 aacagcgcca cgcccgcagt tccgcaaata gcccccagga ccgccatcaa tcgtatcggg
    13741 ctacctagca gagcggcaga gatgaacacg accatcagcg gctgcacagc gcctaccgtc
    13801 gccgcgaccc cgcccggcag gcggtagacc gaaataaaca acaagctcca gaatagcgaa
    13861 atattaagtg cgccgaggat gaagatgcgc atccaccaga ttcccgttgg aatctgtcgg
    13921 acgatcatca cgagcaataa acccgccggc aacgcccgca gcagcatacc ggcgacccct
    13981 cggcctcgct gttcgggctc cacgaaaacg ccggacagat gcgccttgtg agcgtccttg
    14041 gggccgtcct cctgtttgaa gaccgacagc ccaatgatct cgccgtcgat gtaggcgccg
    14101 aatgccacgg catctcgcaa ccgttcagcg aacgcctcca tgggcttttt ctcctcgtgc
    14161 tcgtaaacgg acccgaacat ctctggagct ttcttcaggg ccgacaatcg gatctcgcgg
    14221 aaatcctgca cgtcggccgc tccaagccgt cgaatctgag ccttaatcac aattgtcaat
    14281 tttaatcctc tgtttatcgg cagttcgtag agcgcgccgt gcgtcccgag cgatactgag
    14341 cgaagcaagt gcgtcgagca gtgcccgctt gttcctgaaa tgccagtaaa gcgctggctg
    14401 ctgaaccccc agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat
    14461 cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga
    14521 cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt
    14581 ccagcttgag cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg
    14641 tcggcgacag cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca
    14701 gcacgacgat ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca
    14761 ggacgcggaa gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga
    14821 agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc
    14881 cattgatcga ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc
    14941 cgataggggt gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg
    15001 cccgcagctc gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct
    15061 tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg
    15121 ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa
    15181 gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc
    15241 tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc
    15301 gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc ctgttcgaga cgacgcgaac
    15361 gctccacggc ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct
    15421 cgatcttggc cgtagcttgc tggaccatcg agccgacgga ctggaaggtt tcgcggggcg
    15481 cacgcatgac ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga
    15541 tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga
    15601 ttgccccgac tcacgccggg gcaatgtgcc cttattcctg atttgacccg cctggtgcct
    15661 tggtgtccag ataatccacc ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt
    15721 ccttctcgta cttggtattc cgaatcttgc cctgcacgaa taccagcgac cccttgccca
    15781 aatacttgcc gtgggcctcg gcctgagagc caaaacactt gatgcggaag aagtcggtgc
    15841 gctcctgctt gtcgccggca tcgttgcgcc acatctaggt actaaaacaa ttcatccagt
    15901 aaaatataat attttatttt ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc
    15961 tcgacatact gttcttcccc gatatcctcc ctgatcgacc ggacgcagaa ggcaatgtca
    16021 taccacttgt ccgccctgcc gcttctccca agatcaataa agccacttac tttgccatct
    16081 ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc
    16141 ttttccgtct ttaaaaaatc atacagctcg cgcggatctt taaatggagt gtcttcttcc
    16201 cagttttcgc aatccacatc ggccagatcg ttattcagta agtaatccaa ttcggctaag
    16261 cggctgtcta agctattcgt atagggacaa tccgatatgt cgatggagtg aaagagcctg
    16321 atgcactccg catacagctc gataatcttt tcagggcttt gttcatcttc atactcttcc
    16381 gagcaaagga cgccatcggc ctcactcatg agcagattgc tccagccatc atgccgttca
    16441 aagtgcagga cctttggaac aggcagcttt ccttccagcc atagcatcat gtccttttcc
    16501 cgttccacat cataggtggt ccctttatac cggctgtccg tcatttttaa atataggttt
    16561 tcattttctc ccaccagctt atatacctta gcaggagaca ttccttccgt atcttttacg
    16621 cagcggtatt tttcgatcag ttttttcaat tccggtgata ttctcatttt agccatttat
    16681 tatttccttc ctcttttcta cagtatttaa agatacccca agaagctaat tataacaaga
    16741 cgaactccaa ttcactgttc cttgcattct aaaaccttaa ataccagaaa acagcttttt
    16801 caaagttgtt ttcaaagttg gcgtataaca tagtatcgac ggagccgatt ttgaaaccac
    16861 aattatgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt tgaggtgctc
    16921 cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg gtggtgcgta
    16981 acggcaaaag caccgccgga catcagcgct atctctgctc tcactgccgt aaaacatggc
    17041 aactgcagtt cacttacacc gcttctcaac ccggtacgca ccagaaaatc attgatatgg
    17101 ccatgaatgg cgttggatgc cgggcaacag cccgcattat gggcgttggc ctcaacacga
    17161 ttttacgtca cttaaaaaac tcaggccgca gtcggtaacc tcgcgcatac agccgggcag
    17221 tgacgtcatc gtctgcgcgg aaatggacga acagtggggc tatgtcgggg ctaaatcgcg
    17281 ccagcgctgg ctgttttacg cgtatgacag tctccggaag acggttgttg cgcacgtatt
    17341 cggtgaacgc actatggcga cgctggggcg tcttatgagc ctgctgtcac cctttgacgt
    17401 ggtgatatgg atgacggatg gctggccgct gtatgaatcc cgcctgaagg gaaagctgca
    17461 cgtaatcagc aagcgatata cgcagcgaat tgagcggcat aacctgaatc tgaggcagca
    17521 cctggcacgg ctgggacgga agtcgctgtc gttctcaaaa tcggtggagc tgcatgacaa
    17581 agtcatcggg cattatctga acataaaaca ctatcaataa gttggagtca ttacccaatt
    17641 atgatagaat ttacaagcta taaggttatt gtcctgggtt tcaagcatta gtccatgcaa
    17701 gtttttatgc tttgcccatt ctatagatat attgataagc gcgctgccta tgccttgccc
    17761 cctgaaatcc ttacatacgg cgatatcttc tatataaaag atatattatc ttatcagtat
    17821 tgtcaatata ttcaaggcaa tctgcctcct catcctcttc atcctcttcg tcttggtagc
    17881 tttttaaata tggcgcttca tagagtaatt ctgtaaaggt ccaattctcg ttttcatacc
    17941 tcggtataat cttacctatc acctcaaatg gttcgctggg tttatcgcac ccccgaacac
    18001 gagcacggca cccgcgacca ctatgccaag aatgcccaag gtaaaaattg ccggccccgc
    18061 catgaagtcc gtgaatgccc cgacggccga agtgaagggc aggccgccac ccaggccgcc
    18121 gccctcactg cccggcacct ggtcgctgaa tgtcgatgcc agcacctgcg gcacgtcaat
    18181 gcttccgggc gtcgcgctcg ggctgatcgc ccatcccgtt actgccccga tcccggcaat
    18241 ggcaaggact gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag
    18301 cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac
    18361 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
    18421 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
    18481 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
    18541 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
    18601 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
    18661 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
    18721 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
    18781 cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt
    18841 atccacaacg ccggcggccg gccgcggtgt ctcgcacacg gcttcgacgg cgtttctggc
    18901 gcgtttgcag ggccatagac ggccgccagc ccagcggcga gggcaaccag cccggtgagc
    18961 gtcggaaagg cgctcggtct tgccttgct
//
