LOCUS       sequence-202702-       18988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101040 (Mel1). 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 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: pSC25
FEATURES             Location/Qualifiers
     source          1..18988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(34..405)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            438..547
                     /note="incP origin of transfer"
     primer_bind     770..789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1023..1040
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1228..1878)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2343..2367
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3575..3599
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3621..3668
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3675..3695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3720..5156,5346..8009)
                     /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          5157..5345
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8010..8057
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8068..8320
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8365..8385
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8740..8815
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8827..8851)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9328..9403
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9415..9439)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9458..9482)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9520..10227
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10777..11122
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11065..11085
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11143..11694
                     /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      11838..12090
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12103..12120)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12103..12119)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12112..12134)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12259..12278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12469..12490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12737..12761
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14357..15505)
                     /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(15804..16598)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16790..17557
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18180..18895
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg ggcggaccac
       61 gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca gcagggcgag
      121 gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag
      181 gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt gctcatagtc
      241 cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg acaggctcat
      301 gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc agtacacctt
      361 gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct tgccctcatc
      421 tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga gcaccgccag
      481 gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta cttcacctat
      541 cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc tttggcaaaa
      601 tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa tgaccccgaa
      661 gcagggttat gcagcggaaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      721 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      781 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      841 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      901 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
      961 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     1021 gcagcgagtc agtgagcgag gaagcggaag agcgccagaa ggccgccaga gaggccgagc
     1081 gcggccgtga ggcttggacg ctagggcagg gcatgaaaaa gcccgtagcg ggctgctacg
     1141 ggcgtctgac gcggtggaaa gggggagggg atgttgtcta catggctctg ctgtagtgag
     1201 tgggttgcgc tccggcagcg gtcctgatca atcgtcaccc tttctcggtc cttcaacgtt
     1261 cctgacaacg agcctccttt tcgccaatcc atcgacaatc accgcgagtc cctgctcgaa
     1321 cgctgcgtcc ggaccggctt cgtcgaaggc gtctatcgcg gcccgcaaca gcggcgagag
     1381 cggagcctgt tcaacggtgc cgccgcgctc gccggcatcg ctgtcgccgg cctgctcctc
     1441 aagcacggcc ccaacagtga agtagctgat tgtcatcagc gcattgacgg cgtccccggc
     1501 cgaaaaaccc gcctcgcaga ggaagcgaag ctgcgcgtcg gccgtttcca tctgcggtgc
     1561 gcccggtcgc gtgccggcat ggatgcgcgc gccatcgcgg taggcgagca gcgcctgcct
     1621 gaagctgcgg gcattcccga tcagaaatga gcgccagtcg tcgtcggctc tcggcaccga
     1681 atgcgtatga ttctccgcca gcatggcttc ggccagtgcg tcgagcagcg cccgcttgtt
     1741 cctgaagtgc cagtaaagcg ccggctgctg aacccccaac cgttccgcca gtttgcgtgt
     1801 cgtcagaccg tctacgccga cctcgttcaa caggtccagg gcggcacgga tcactgtatt
     1861 cggctgcaac tttgtcatgc ttgacacttt atcactgata aacataatat gtccaccaac
     1921 ttatcagtga taaagaatcc gcgcgttcaa tcggaccagc ggaggctggt ccggaggcca
     1981 gacgtgaaac ccaacatacc cctgatcgta attctgagca ctgtcgcgct cgacgctgtc
     2041 ggcatcggcc tgattatgcc ggtgctgccg ggcctcctgc gcgatctggt tcactcgaac
     2101 gacgtcaccg cccactatgg cattctgctg gcgctgtatg cgttggtgca atttgcctgc
     2161 gcacctgtgc tgggcgcgct gtcggatcgt ttcgggcggc ggccaatctt gctcgtctcg
     2221 ctggccggcg ccagatctgg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg
     2281 gataaacctt ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt
     2341 aggtttaccc gccaatatat cctgtcaaac actgatagtt taaactgaag gcgggaaacg
     2401 acaatctgat catgagcgga gaattaaggg agtcacgtta tgacccccgc cgatgacgcg
     2461 ggacaagccg ttttacgttt ggaactgaca gaaccgcaac gttgaaggag ccactcagcc
     2521 gcgggtttct ggagtttaat gagctaagca catacgtcag aaaccattat tgcgcgttca
     2581 aaagtcgcct aaggtcacta tcagctagac ggacgcggcc gcgggcccaa tataacaacg
     2641 acgtcgtaac agataaagcg aagcttgaag gtgcatgtga ctccgtcaag attacgaaac
     2701 cgccaactac cacgcaaatt gcaattctca atttcctaga aggactctcc gaaaatgcat
     2761 ccaataccaa atattacccg tgtcataggc accaagtgac accatacatg aacacgcgtc
     2821 acaatatgac tggagaaggg ttccacacct tatgctataa aacgccccac acccctcctc
     2881 cttccttcgc agttcaattc caatatattc cattctctct gtgtatttcc ctacctctcc
     2941 cttcaaggtt agtcgatttc ttctgttttt cttcttcgtt ctttccatga attgtgtatg
     3001 ttctttgatc aatacgatgt tgatttgatt gtgttttgtt tggtttcatc gatcttcaat
     3061 tttcataatc agattcagct tttattatct ttacaacaac gtccttaatt tgatgattct
     3121 ttaatcgtag atttgctcta attagagctt tttcatgtca gatcccttta caacaagcct
     3181 taattgttga ttcattaatc gtagattagg gcttttttca ttgattactt cagatccgtt
     3241 aaacgtaacc atagatcagg gctttttcat gaattacttc agatccgtta aacaacagcc
     3301 ttatttttta tacttctgtg gtttttcaag aaattgttca gatccgttga caaaaagcct
     3361 tattcgttga ttctatatcg tttttcgaga gatattgctc agatctgtta gcaactgcct
     3421 tgtttgttga ttctattgcc gtggattagg gttttttttc acgagattgc ttcagatccg
     3481 tacttaagat tacgtaatgg attttgattc tgatttatct gtgattgttg actcgacagg
     3541 taccttcaaa cctcgaggcg cgccaagcta tcaaacaagt ttgtacaaaa aagcaggctc
     3601 cgaattcgcc cttcaccatg gattacaagg atgatgatga taaggattac aaggatgatg
     3661 atgataagat ggctccaaag aagaagagaa aggttggaat ccacggagtt ccagctgctg
     3721 ataagaagta ctctatcgga cttgacatcg gaaccaactc tgttggatgg gctgttatca
     3781 ccgatgagta caaggttcca tctaagaagt tcaaggttct tggaaacacc gatagacact
     3841 ctatcaagaa gaaccttatc ggtgctcttc ttttcgattc tggagagacc gctgaggcta
     3901 ccagattgaa gagaaccgct agaagaagat acaccagaag aaagaacaga atctgctacc
     3961 ttcaggaaat cttctctaac gagatggcta aggttgatga ttctttcttc cacagacttg
     4021 aggagtcttt ccttgttgag gaggataaga agcacgagag acacccaatc ttcggaaaca
     4081 tcgttgatga ggttgcttac cacgagaagt acccaaccat ctaccacctt agaaagaagt
     4141 tggttgattc taccgataag gctgatctta gacttatcta ccttgctctt gctcacatga
     4201 tcaagttcag aggacacttc cttatcgagg gagaccttaa cccagataac tctgatgttg
     4261 ataagttgtt catccagctt gttcagacct acaaccagct tttcgaggag aacccaatca
     4321 acgcttctgg agttgatgct aaggctatcc tttctgctag actttctaag tctcgtagac
     4381 ttgagaacct tatcgctcag cttccaggag agaagaagaa cggacttttc ggaaacctta
     4441 tcgctctttc tcttggactt accccaaact tcaagtctaa cttcgatctt gctgaggatg
     4501 ctaagttgca gctttctaag gatacctacg atgatgatct tgataacctt cttgctcaga
     4561 tcggagatca gtacgctgat cttttccttg ctgctaagaa cctttctgat gctatccttc
     4621 tttctgacat ccttagagtt aacaccgaga tcaccaaggc tccactttct gcttctatga
     4681 tcaagagata cgatgagcac caccaggatc ttaccctttt gaaggctctt gttagacagc
     4741 agcttccaga gaagtacaag gaaatcttct tcgatcagtc taagaacgga tacgctggat
     4801 acatcgatgg aggagcttct caggaggagt tctacaagtt catcaagcca atccttgaga
     4861 agatggatgg aaccgaggag cttcttgtta agttgaacag agaggatctt cttagaaagc
     4921 agagaacctt cgataacgga tctatcccac accagatcca ccttggagag cttcacgcta
     4981 tccttcgtag acaggaggat ttctacccat tcttgaagga taacagagag aagatcgaga
     5041 agatccttac cttcagaatc ccatactacg ttggaccact tgctagagga aactctcgtt
     5101 tcgcttggat gaccagaaag tctgaggaga ccatcacccc ttggaacttc gaggaggtaa
     5161 gtttctgctt ctacctttga tatatatata ataattatca ttaattagta gtaatataat
     5221 atttcaaata tttttttcaa aataaaagaa tgtagtatat agcaattgct tttctgtagt
     5281 ttataagtgt gtatatttta atttataact tttctaatat atgaccaaaa tttgttgatg
     5341 tgcaggttgt tgataaggga gcttctgctc agtctttcat cgagagaatg accaacttcg
     5401 ataagaacct tccaaacgag aaggttcttc caaagcactc tcttctttac gagtacttca
     5461 ccgtttacaa cgagcttacc aaggttaagt acgttaccga gggaatgaga aagccagctt
     5521 tcctttctgg agagcagaag aaggctatcg ttgatcttct tttcaagacc aacagaaagg
     5581 ttaccgttaa gcagttgaag gaggattact tcaagaagat cgagtgcttc gattctgttg
     5641 aaatctctgg agttgaggat agattcaacg cttctcttgg aacctaccac gatcttttga
     5701 agatcatcaa ggataaggat ttccttgata acgaggagaa cgaggacatc cttgaggaca
     5761 tcgttcttac ccttaccctt ttcgaggata gagagatgat cgaggagaga ctcaagacct
     5821 acgctcacct tttcgatgat aaggttatga agcagttgaa gagaagaaga tacaccggat
     5881 ggggtagact ttctcgtaag ttgatcaacg gaatcagaga taagcagtct ggaaagacca
     5941 tccttgattt cttgaagtct gatggattcg ctaacagaaa cttcatgcag cttatccacg
     6001 atgattctct taccttcaag gaggacatcc agaaggctca ggtttctgga cagggagatt
     6061 ctcttcacga gcacatcgct aaccttgctg gatctccagc tatcaagaag ggaatccttc
     6121 agaccgttaa ggttgttgat gagcttgtta aggttatggg tagacacaag ccagagaaca
     6181 tcgttatcga gatggctaga gagaaccaga ccacccagaa gggacagaag aactctcgtg
     6241 agagaatgaa gagaatcgag gagggaatca aggagcttgg atctcaaatc ttgaaggagc
     6301 acccagttga gaacacccag cttcagaacg agaagttgta cctttactac cttcagaacg
     6361 gaagagatat gtacgttgat caggagcttg acatcaacag actttctgat tacgatgttg
     6421 atcacatcgt tccacagtct ttcttgaagg atgattctat cgataacaag gttcttaccc
     6481 gttctgataa gaacagagga aagtctgata acgttccatc tgaggaggtt gttaagaaga
     6541 tgaagaacta ctggagacag cttcttaacg ctaagttgat cacccagaga aagttcgata
     6601 accttaccaa ggctgagaga ggaggacttt ctgagcttga taaggctgga ttcatcaaga
     6661 gacagcttgt tgagaccaga cagatcacca agcacgttgc tcagatcctt gattctcgta
     6721 tgaacaccaa gtacgatgag aacgataagt tgatcagaga ggttaaggtt atcaccttga
     6781 agtctaagtt ggtttctgat ttcagaaagg atttccagtt ctacaaggtt agagagatca
     6841 acaactacca ccacgctcac gatgcttacc ttaacgctgt tgttggaacc gctcttatca
     6901 agaagtaccc aaagttggag tctgagttcg tttacggaga ttacaaggtt tacgatgtta
     6961 gaaagatgat cgctaagtct gagcaggaga tcggaaaggc taccgctaag tacttcttct
     7021 actctaacat catgaacttc ttcaagaccg agatcaccct tgctaacgga gagatcagaa
     7081 agagaccact tatcgagacc aacggagaga ccggagagat cgtttgggat aagggaagag
     7141 atttcgctac cgttagaaag gttctttcta tgccacaggt taacatcgtt aagaaaaccg
     7201 aggttcagac cggaggattc tctaaggagt ctatccttcc aaagagaaac tctgataagt
     7261 tgatcgctag aaagaaggat tgggacccaa agaagtacgg aggattcgat tctccaaccg
     7321 ttgcttactc tgttcttgtt gttgctaagg ttgagaaggg aaagtctaag aagttgaagt
     7381 ctgttaagga gcttcttgga atcaccatca tggagcgttc ttctttcgag aagaacccaa
     7441 tcgatttcct tgaggctaag ggatacaagg aggttaagaa ggatcttatc atcaagttgc
     7501 caaagtactc tcttttcgag cttgagaacg gaagaaagag aatgcttgct tctgctggag
     7561 agcttcagaa gggaaacgag cttgctcttc catctaagta cgttaacttc ctttaccttg
     7621 cttctcacta cgagaagttg aagggatctc cagaggataa cgagcagaag cagcttttcg
     7681 ttgagcagca caagcactac cttgatgaga tcatcgagca aatctctgag ttctctaaga
     7741 gagttatcct tgctgatgct aaccttgata aggttctttc tgcttacaac aagcacagag
     7801 ataagccaat cagagagcag gctgagaaca tcatccacct tttcaccctt accaaccttg
     7861 gtgctccagc tgctttcaag tacttcgata ccaccatcga tagaaaaaga tacacctcta
     7921 ccaaggaggt tcttgatgct acccttatcc accagtctat caccggactt tacgagacca
     7981 gaatcgatct ttctcagctt ggaggagata agagaccagc tgctaccaag aaggctggac
     8041 aggctaagaa gaagaagtga gacgtccgat cgttcaaaca tttggcaata aagtttctta
     8101 agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt
     8161 aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt
     8221 agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag
     8281 gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattgat cccccctcga
     8341 cagcttccgg aaagggcgaa ttcgcaactt tgtatacaaa agttgccgag ctcgcggccg
     8401 catgttgttg ttaccagaaa gtaaataaat gttcaatctc tgatgttctc aagtaagtga
     8461 gttttattgg gaataatatt aacttatgtt cttcttgcat ttgatttctt tgccgctctc
     8521 ttcttctatc ttaaatctgt gtatactatt tcactattgg gctttttatt agtctataat
     8581 gggactcaaa ataaggcttt ggcccacatc aaaaagataa gtcacaaatc aaaactaaat
     8641 tcagagtctt ttctcccaca tcggtcactg tactcatttt gtgtttgttt atatattaca
     8701 cgaaccgatc tttggtacgg gacattttta tgcgtccacg ttttagagct agaaatagca
     8761 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     8821 ttctagaccc agctttcttg tacaaagttg gcattaccat ggcttcgttg aacaacggaa
     8881 actcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct tcttcttcgt
     8941 tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct ttcgttttct
     9001 tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc ccaggattag
     9061 aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca ttcttaagat
     9121 atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag gcccatttat
     9181 atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac aatcttcaaa
     9241 agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta gagtcgaagt
     9301 agtgattgtg gatgataatc cgatctcgtt ttagagctag aaatagcaag ttaaaataag
     9361 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctagacccag
     9421 ctttcttgta caaagttggc attactgcag gtcgacgacc cagctttctt gtacaaagtg
     9481 gttcgataat tcttaattaa ctagttctaa tctagagtcc tgctttaatg agatatgcga
     9541 gacgcctatg atcgcatgat atttgctttc aattctgttg tgcacgttgt aaaaaacctg
     9601 agcatgtgta gctcagatcc ttaccgccgg tttcggttca ttctaatgaa tatatcaccc
     9661 gttactatcg tatttttatg aataatattc tccgttcaat ttactgattg taccctacta
     9721 cttatatgta caatattaaa atgaaaacaa tatattgtgc tgaataggtt tatagcgaca
     9781 tctatgatag agcgccacaa taacaaacaa ttgcgtttta ttattacaaa tccaatttta
     9841 aaaaaagcgg cagaaccggt caaacctaaa agactgatta cataaatctt attcaaattt
     9901 caaaagtgcc ccaggggcta gtatctacga cacaccgagc ggcgaactaa taacgctcac
     9961 tgaagggaac tccggttccc cgccggcgcg catgggtgag attccttgaa gttgagtatt
    10021 ggccgtccgc tctaccgaaa gttacgggca ccattcaacc cggtccagca cggcggccgg
    10081 gtaaccgact tgctgccccg agaattatgc agcatttttt tggtgtatgt gggccccaaa
    10141 tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt gggcgggtcc agggcgaatt
    10201 ttgcgacaac atgtcgaggc tcagcaggac ctgcaggcat gcaagctagt gaagaatcga
    10261 tttgctacgt attgcccctg caggtcccca gattagcctt ttcaatttca gaaagaatgc
    10321 taacccacag atggttagag aggcttacgc agcaggtctc atcaagacga tctacccgag
    10381 caataatctc caggaaatca aataccttcc caagaaggtt aaagatgcag tcaaaagatt
    10441 caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca gaagtactat
    10501 tccagtatgg acgattcaag gcttgcttca caaaccaagg caagtaatag agattggagt
    10561 ctctaaaaag gtagttccca ctgaatcaaa ggccatggag tcaaagattc aaatagagga
    10621 cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct tacgactcaa
    10681 tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct actccaaaaa
    10741 tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac aaagggtaat
    10801 atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg tgaagatagt
    10861 ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ccatcgttga
    10921 agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga
    10981 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
    11041 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
    11101 ttcatttcat ttggagagaa cacgggggac tctagaggat ccatgagccc agaacgacgc
    11161 ccggccgaca tccgccgtgc caccgaggcg gacatgccgg cggtctgcac catcgtcaac
    11221 cactacatcg agacaagcac ggtcaacttc cgtaccgagc cgcaggaacc gcaggagtgg
    11281 acggacgacc tcgtccgtct gcgggagcgc tatccctggc tcgtcgccga ggtggacggc
    11341 gaggtcgccg gcatcgccta cgcgggcccc tggaaggcac gcaacgccta cgactggacg
    11401 gccgagtcga ccgtgtacgt ctccccccgc caccagcgga cgggactggg ctccacgctc
    11461 tacacccacc tgctgaagtc cctggaggca cagggcttca agagcgtggt cgctgtcatc
    11521 gggctgccca acgacccgag cgtgcgcatg cacgaggcgc tcggatatgc cccccgcggc
    11581 atgctgcggg cggccggctt caagcacggg aactggcatg acgtgggttt ctggcagctg
    11641 gacttcagcc tgccggtacc gccccgtccg gtcctgcccg tcaccgagat ctgaacggag
    11701 tgcgcgtggc atcgcccgag ttggagctgg tacgggaact catcgaactc aactggcata
    11761 cccgcaatgg tgaggtggaa ccgcggcgga tcgcgtacga ccgtgcccag gagggggtac
    11821 cgagctcgaa tttccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc
    11881 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
    11941 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
    12001 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
    12061 cgcgcgcggt gtcatctatg ttactagatc gggaattaat tcactggccg tcgttttaca
    12121 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
    12181 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
    12241 cagcctgaat ggcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    12301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    12361 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
    12421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    12481 tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
    12541 tgccgatttc ggaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
    12601 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
    12661 actggtgaaa agaaaaacca ccccagtaca ttaaaaacgt ccgcaatgtg ttattaagtt
    12721 gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc
    12781 ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccgggacg
    12841 gcgtcagcgg gagagccgtt gtaaggcggc agactttgct catgttaccg atgctattcg
    12901 gaagaacggc aactaagctg ccgggtttga aacacggatg atctcgcgga gggtagcatg
    12961 ttgattgtaa cgatgacaga gcgttgctgc ctgtgatcaa atatcatctc cctcgcagag
    13021 atccgaatta tcagccttct tattcatttc tcgcttaacc gtgacaggct gtcgatcttg
    13081 agaactatgc cgacataata ggaaatcgct ggataaagcc gctgaggaag ctgagtggcg
    13141 ctatttcttt agaagtgaac gttgacgata tcaactcccc tatccattgc tcaccgaatg
    13201 gtacaggtcg gggacccgaa gttccgactg tcggcctgat gcatccccgg ctgatcgacc
    13261 ccagatctgg ggctgagaaa gcccagtaag gaaacaactg taggttcgag tcgcgagatc
    13321 ccccggaacc aaaggaagta ggttaaaccc gctccgatca ggccgagcca cgccaggccg
    13381 agaacattgg ttcctgtagg catcgggatt ggcggatcaa acactaaagc tactggaacg
    13441 agcagaagtc ctccggccgc cagttgccag gcggtaaagg tgagcagagg cacgggaggt
    13501 tgccacttgc gggtcagcac ggttccgaac gccatggaaa ccgcccccgc caggcccgct
    13561 gcgacgccga caggatctag cgctgcgttt ggtgtcaaca ccaacagcgc cacgcccgca
    13621 gttccgcaaa tagcccccag gaccgccatc aatcgtatcg ggctacctag cagagcggca
    13681 gagatgaaca cgaccatcag cggctgcaca gcgcctaccg tcgccgcgac cccgcccggc
    13741 aggcggtaga ccgaaataaa caacaagctc cagaatagcg aaatattaag tgcgccgagg
    13801 atgaagatgc gcatccacca gattcccgtt ggaatctgtc ggacgatcat cacgagcaat
    13861 aaacccgccg gcaacgcccg cagcagcata ccggcgaccc ctcggcctcg ctgttcgggc
    13921 tccacgaaaa cgccggacag atgcgccttg tgagcgtcct tggggccgtc ctcctgtttg
    13981 aagaccgaca gcccaatgat ctcgccgtcg atgtaggcgc cgaatgccac ggcatctcgc
    14041 aaccgttcag cgaacgcctc catgggcttt ttctcctcgt gctcgtaaac ggacccgaac
    14101 atctctggag ctttcttcag ggccgacaat cggatctcgc ggaaatcctg cacgtcggcc
    14161 gctccaagcc gtcgaatctg agccttaatc acaattgtca attttaatcc tctgtttatc
    14221 ggcagttcgt agagcgcgcc gtgcgtcccg agcgatactg agcgaagcaa gtgcgtcgag
    14281 cagtgcccgc ttgttcctga aatgccagta aagcgctggc tgctgaaccc ccagccggaa
    14341 ctgaccccac aaggccctag cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt
    14401 ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg cgccacttct
    14461 tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt ttccagcttg agcgggtacg
    14521 gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt
    14581 acttctccca tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt
    14641 cgatcaggac ctggcaacgg gacgttttct tgccacggtc caggacgcgg aagcggtgca
    14701 gcagcgacac cgattccagg tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct
    14761 gtaggcgcga caggcattcc tcggccttcg tgtaataccg gccattgatc gaccagccca
    14821 ggtcctggca aagctcgtag aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg
    14881 cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg
    14941 tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc
    15001 gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg
    15061 ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga aagctgcatt tccttgatct
    15121 gctgcttcgt gtgtttcagc aacgcggcct gcttggcctc gctgacctgt tttgccaggt
    15181 cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg
    15241 acttcgccaa acctgccgcc tcctgttcga gacgacgcga acgctccacg gcggccgatg
    15301 gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt
    15361 gctggaccat cgagccgacg gactggaagg tttcgcgggg cgcacgcatg acggtgcggc
    15421 ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc gatcagttct tgcctgtatg
    15481 ccttccggtc aaacgtccga ttcattcacc ctccttgcgg gattgccccg actcacgccg
    15541 gggcaatgtg cccttattcc tgatttgacc cgcctggtgc cttggtgtcc agataatcca
    15601 ccttatcggc aatgaagtcg gtcccgtaga ccgtctggcc gtccttctcg tacttggtat
    15661 tccgaatctt gccctgcacg aataccagcg accccttgcc caaatacttg ccgtgggcct
    15721 cggcctgaga gccaaaacac ttgatgcgga agaagtcggt gcgctcctgc ttgtcgccgg
    15781 catcgttgcg ccacatctag gtactaaaac aattcatcca gtaaaatata atattttatt
    15841 ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc
    15901 ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg
    15961 ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg
    16021 tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa
    16081 tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca
    16141 tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc
    16201 gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc
    16261 tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg
    16321 gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga
    16381 acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg
    16441 gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc
    16501 ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc
    16561 agttttttca attccggtga tattctcatt ttagccattt attatttcct tcctcttttc
    16621 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt
    16681 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt
    16741 tggcgtataa catagtatcg acggagccga ttttgaaacc acaattatgg gtgatgctgc
    16801 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
    16861 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
    16921 gacatcagcg ctatctctgc tctcactgcc gtaaaacatg gcaactgcag ttcacttaca
    16981 ccgcttctca acccggtacg caccagaaaa tcattgatat ggccatgaat ggcgttggat
    17041 gccgggcaac agcccgcatt atgggcgttg gcctcaacac gattttacgt cacttaaaaa
    17101 actcaggccg cagtcggtaa cctcgcgcat acagccgggc agtgacgtca tcgtctgcgc
    17161 ggaaatggac gaacagtggg gctatgtcgg ggctaaatcg cgccagcgct ggctgtttta
    17221 cgcgtatgac agtctccgga agacggttgt tgcgcacgta ttcggtgaac gcactatggc
    17281 gacgctgggg cgtcttatga gcctgctgtc accctttgac gtggtgatat ggatgacgga
    17341 tggctggccg ctgtatgaat cccgcctgaa gggaaagctg cacgtaatca gcaagcgata
    17401 tacgcagcga attgagcggc ataacctgaa tctgaggcag cacctggcac ggctgggacg
    17461 gaagtcgctg tcgttctcaa aatcggtgga gctgcatgac aaagtcatcg ggcattatct
    17521 gaacataaaa cactatcaat aagttggagt cattacccaa ttatgataga atttacaagc
    17581 tataaggtta ttgtcctggg tttcaagcat tagtccatgc aagtttttat gctttgccca
    17641 ttctatagat atattgataa gcgcgctgcc tatgccttgc cccctgaaat ccttacatac
    17701 ggcgatatct tctatataaa agatatatta tcttatcagt attgtcaata tattcaaggc
    17761 aatctgcctc ctcatcctct tcatcctctt cgtcttggta gctttttaaa tatggcgctt
    17821 catagagtaa ttctgtaaag gtccaattct cgttttcata cctcggtata atcttaccta
    17881 tcacctcaaa tggttcgctg ggtttatcgc acccccgaac acgagcacgg cacccgcgac
    17941 cactatgcca agaatgccca aggtaaaaat tgccggcccc gccatgaagt ccgtgaatgc
    18001 cccgacggcc gaagtgaagg gcaggccgcc acccaggccg ccgccctcac tgcccggcac
    18061 ctggtcgctg aatgtcgatg ccagcacctg cggcacgtca atgcttccgg gcgtcgcgct
    18121 cgggctgatc gcccatcccg ttactgcccc gatcccggca atggcaagga ctgccagcgc
    18181 tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag
    18241 gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg
    18301 gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag
    18361 caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg
    18421 attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac
    18481 tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt
    18541 gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt
    18601 aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc
    18661 gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt
    18721 ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc
    18781 cggccgcggt gtctcgcaca cggcttcgac ggcgtttctg gcgcgtttgc agggccatag
    18841 acggccgcca gcccagcggc gagggcaacc agcccggtga gcgtcggaaa ggcgctcggt
    18901 cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga
    18961 gcgcatgggg acgtgcttgg caatcacg
//
