LOCUS       sequence-202753-       18989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr4g020620 (Pho2ab). Also 
            expresses Cas9 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: pSC27
FEATURES             Location/Qualifiers
     source          1..18989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(64..435)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            468..577
                     /note="incP origin of transfer"
     primer_bind     800..819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1053..1070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1258..1908)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2373..2397
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3605..3629
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3651..3698
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3705..3725
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3750..5186,5376..8039)
                     /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          5187..5375
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8040..8087
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8098..8350
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8395..8415
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8770..8845
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8857..8881)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9359..9434
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9446..9470)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9489..9513)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9551..10258
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10808..11153
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11096..11116
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11174..11725
                     /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      11869..12121
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12134..12151)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12134..12150)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12143..12165)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12290..12309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12500..12521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12768..12792
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14388..15536)
                     /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(15835..16629)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16821..17588
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18211..18926
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa
       61 aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa gctcgtcctg
      121 cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg
      181 cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt cgccattgat
      241 gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc
      301 gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt cctcaatcgc
      361 tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt
      421 ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg
      481 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
      541 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
      601 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
      661 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa agcgccacgc
      721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1081 agcgccagaa ggccgccaga gaggccgagc gcggccgtga ggcttggacg ctagggcagg
     1141 gcatgaaaaa gcccgtagcg ggctgctacg ggcgtctgac gcggtggaaa gggggagggg
     1201 atgttgtcta catggctctg ctgtagtgag tgggttgcgc tccggcagcg gtcctgatca
     1261 atcgtcaccc tttctcggtc cttcaacgtt cctgacaacg agcctccttt tcgccaatcc
     1321 atcgacaatc accgcgagtc cctgctcgaa cgctgcgtcc ggaccggctt cgtcgaaggc
     1381 gtctatcgcg gcccgcaaca gcggcgagag cggagcctgt tcaacggtgc cgccgcgctc
     1441 gccggcatcg ctgtcgccgg cctgctcctc aagcacggcc ccaacagtga agtagctgat
     1501 tgtcatcagc gcattgacgg cgtccccggc cgaaaaaccc gcctcgcaga ggaagcgaag
     1561 ctgcgcgtcg gccgtttcca tctgcggtgc gcccggtcgc gtgccggcat ggatgcgcgc
     1621 gccatcgcgg taggcgagca gcgcctgcct gaagctgcgg gcattcccga tcagaaatga
     1681 gcgccagtcg tcgtcggctc tcggcaccga atgcgtatga ttctccgcca gcatggcttc
     1741 ggccagtgcg tcgagcagcg cccgcttgtt cctgaagtgc cagtaaagcg ccggctgctg
     1801 aacccccaac cgttccgcca gtttgcgtgt cgtcagaccg tctacgccga cctcgttcaa
     1861 caggtccagg gcggcacgga tcactgtatt cggctgcaac tttgtcatgc ttgacacttt
     1921 atcactgata aacataatat gtccaccaac ttatcagtga taaagaatcc gcgcgttcaa
     1981 tcggaccagc ggaggctggt ccggaggcca gacgtgaaac ccaacatacc cctgatcgta
     2041 attctgagca ctgtcgcgct cgacgctgtc ggcatcggcc tgattatgcc ggtgctgccg
     2101 ggcctcctgc gcgatctggt tcactcgaac gacgtcaccg cccactatgg cattctgctg
     2161 gcgctgtatg cgttggtgca atttgcctgc gcacctgtgc tgggcgcgct gtcggatcgt
     2221 ttcgggcggc ggccaatctt gctcgtctcg ctggccggcg ccagatctgg ggaaccctgt
     2281 ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc
     2341 cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac
     2401 actgatagtt taaactgaag gcgggaaacg acaatctgat catgagcgga gaattaaggg
     2461 agtcacgtta tgacccccgc cgatgacgcg ggacaagccg ttttacgttt ggaactgaca
     2521 gaaccgcaac gttgaaggag ccactcagcc gcgggtttct ggagtttaat gagctaagca
     2581 catacgtcag aaaccattat tgcgcgttca aaagtcgcct aaggtcacta tcagctagac
     2641 ggacgcggcc gcgggcccaa tataacaacg acgtcgtaac agataaagcg aagcttgaag
     2701 gtgcatgtga ctccgtcaag attacgaaac cgccaactac cacgcaaatt gcaattctca
     2761 atttcctaga aggactctcc gaaaatgcat ccaataccaa atattacccg tgtcataggc
     2821 accaagtgac accatacatg aacacgcgtc acaatatgac tggagaaggg ttccacacct
     2881 tatgctataa aacgccccac acccctcctc cttccttcgc agttcaattc caatatattc
     2941 cattctctct gtgtatttcc ctacctctcc cttcaaggtt agtcgatttc ttctgttttt
     3001 cttcttcgtt ctttccatga attgtgtatg ttctttgatc aatacgatgt tgatttgatt
     3061 gtgttttgtt tggtttcatc gatcttcaat tttcataatc agattcagct tttattatct
     3121 ttacaacaac gtccttaatt tgatgattct ttaatcgtag atttgctcta attagagctt
     3181 tttcatgtca gatcccttta caacaagcct taattgttga ttcattaatc gtagattagg
     3241 gcttttttca ttgattactt cagatccgtt aaacgtaacc atagatcagg gctttttcat
     3301 gaattacttc agatccgtta aacaacagcc ttatttttta tacttctgtg gtttttcaag
     3361 aaattgttca gatccgttga caaaaagcct tattcgttga ttctatatcg tttttcgaga
     3421 gatattgctc agatctgtta gcaactgcct tgtttgttga ttctattgcc gtggattagg
     3481 gttttttttc acgagattgc ttcagatccg tacttaagat tacgtaatgg attttgattc
     3541 tgatttatct gtgattgttg actcgacagg taccttcaaa cctcgaggcg cgccaagcta
     3601 tcaaacaagt ttgtacaaaa aagcaggctc cgaattcgcc cttcaccatg gattacaagg
     3661 atgatgatga taaggattac aaggatgatg atgataagat ggctccaaag aagaagagaa
     3721 aggttggaat ccacggagtt ccagctgctg ataagaagta ctctatcgga cttgacatcg
     3781 gaaccaactc tgttggatgg gctgttatca ccgatgagta caaggttcca tctaagaagt
     3841 tcaaggttct tggaaacacc gatagacact ctatcaagaa gaaccttatc ggtgctcttc
     3901 ttttcgattc tggagagacc gctgaggcta ccagattgaa gagaaccgct agaagaagat
     3961 acaccagaag aaagaacaga atctgctacc ttcaggaaat cttctctaac gagatggcta
     4021 aggttgatga ttctttcttc cacagacttg aggagtcttt ccttgttgag gaggataaga
     4081 agcacgagag acacccaatc ttcggaaaca tcgttgatga ggttgcttac cacgagaagt
     4141 acccaaccat ctaccacctt agaaagaagt tggttgattc taccgataag gctgatctta
     4201 gacttatcta ccttgctctt gctcacatga tcaagttcag aggacacttc cttatcgagg
     4261 gagaccttaa cccagataac tctgatgttg ataagttgtt catccagctt gttcagacct
     4321 acaaccagct tttcgaggag aacccaatca acgcttctgg agttgatgct aaggctatcc
     4381 tttctgctag actttctaag tctcgtagac ttgagaacct tatcgctcag cttccaggag
     4441 agaagaagaa cggacttttc ggaaacctta tcgctctttc tcttggactt accccaaact
     4501 tcaagtctaa cttcgatctt gctgaggatg ctaagttgca gctttctaag gatacctacg
     4561 atgatgatct tgataacctt cttgctcaga tcggagatca gtacgctgat cttttccttg
     4621 ctgctaagaa cctttctgat gctatccttc tttctgacat ccttagagtt aacaccgaga
     4681 tcaccaaggc tccactttct gcttctatga tcaagagata cgatgagcac caccaggatc
     4741 ttaccctttt gaaggctctt gttagacagc agcttccaga gaagtacaag gaaatcttct
     4801 tcgatcagtc taagaacgga tacgctggat acatcgatgg aggagcttct caggaggagt
     4861 tctacaagtt catcaagcca atccttgaga agatggatgg aaccgaggag cttcttgtta
     4921 agttgaacag agaggatctt cttagaaagc agagaacctt cgataacgga tctatcccac
     4981 accagatcca ccttggagag cttcacgcta tccttcgtag acaggaggat ttctacccat
     5041 tcttgaagga taacagagag aagatcgaga agatccttac cttcagaatc ccatactacg
     5101 ttggaccact tgctagagga aactctcgtt tcgcttggat gaccagaaag tctgaggaga
     5161 ccatcacccc ttggaacttc gaggaggtaa gtttctgctt ctacctttga tatatatata
     5221 ataattatca ttaattagta gtaatataat atttcaaata tttttttcaa aataaaagaa
     5281 tgtagtatat agcaattgct tttctgtagt ttataagtgt gtatatttta atttataact
     5341 tttctaatat atgaccaaaa tttgttgatg tgcaggttgt tgataaggga gcttctgctc
     5401 agtctttcat cgagagaatg accaacttcg ataagaacct tccaaacgag aaggttcttc
     5461 caaagcactc tcttctttac gagtacttca ccgtttacaa cgagcttacc aaggttaagt
     5521 acgttaccga gggaatgaga aagccagctt tcctttctgg agagcagaag aaggctatcg
     5581 ttgatcttct tttcaagacc aacagaaagg ttaccgttaa gcagttgaag gaggattact
     5641 tcaagaagat cgagtgcttc gattctgttg aaatctctgg agttgaggat agattcaacg
     5701 cttctcttgg aacctaccac gatcttttga agatcatcaa ggataaggat ttccttgata
     5761 acgaggagaa cgaggacatc cttgaggaca tcgttcttac ccttaccctt ttcgaggata
     5821 gagagatgat cgaggagaga ctcaagacct acgctcacct tttcgatgat aaggttatga
     5881 agcagttgaa gagaagaaga tacaccggat ggggtagact ttctcgtaag ttgatcaacg
     5941 gaatcagaga taagcagtct ggaaagacca tccttgattt cttgaagtct gatggattcg
     6001 ctaacagaaa cttcatgcag cttatccacg atgattctct taccttcaag gaggacatcc
     6061 agaaggctca ggtttctgga cagggagatt ctcttcacga gcacatcgct aaccttgctg
     6121 gatctccagc tatcaagaag ggaatccttc agaccgttaa ggttgttgat gagcttgtta
     6181 aggttatggg tagacacaag ccagagaaca tcgttatcga gatggctaga gagaaccaga
     6241 ccacccagaa gggacagaag aactctcgtg agagaatgaa gagaatcgag gagggaatca
     6301 aggagcttgg atctcaaatc ttgaaggagc acccagttga gaacacccag cttcagaacg
     6361 agaagttgta cctttactac cttcagaacg gaagagatat gtacgttgat caggagcttg
     6421 acatcaacag actttctgat tacgatgttg atcacatcgt tccacagtct ttcttgaagg
     6481 atgattctat cgataacaag gttcttaccc gttctgataa gaacagagga aagtctgata
     6541 acgttccatc tgaggaggtt gttaagaaga tgaagaacta ctggagacag cttcttaacg
     6601 ctaagttgat cacccagaga aagttcgata accttaccaa ggctgagaga ggaggacttt
     6661 ctgagcttga taaggctgga ttcatcaaga gacagcttgt tgagaccaga cagatcacca
     6721 agcacgttgc tcagatcctt gattctcgta tgaacaccaa gtacgatgag aacgataagt
     6781 tgatcagaga ggttaaggtt atcaccttga agtctaagtt ggtttctgat ttcagaaagg
     6841 atttccagtt ctacaaggtt agagagatca acaactacca ccacgctcac gatgcttacc
     6901 ttaacgctgt tgttggaacc gctcttatca agaagtaccc aaagttggag tctgagttcg
     6961 tttacggaga ttacaaggtt tacgatgtta gaaagatgat cgctaagtct gagcaggaga
     7021 tcggaaaggc taccgctaag tacttcttct actctaacat catgaacttc ttcaagaccg
     7081 agatcaccct tgctaacgga gagatcagaa agagaccact tatcgagacc aacggagaga
     7141 ccggagagat cgtttgggat aagggaagag atttcgctac cgttagaaag gttctttcta
     7201 tgccacaggt taacatcgtt aagaaaaccg aggttcagac cggaggattc tctaaggagt
     7261 ctatccttcc aaagagaaac tctgataagt tgatcgctag aaagaaggat tgggacccaa
     7321 agaagtacgg aggattcgat tctccaaccg ttgcttactc tgttcttgtt gttgctaagg
     7381 ttgagaaggg aaagtctaag aagttgaagt ctgttaagga gcttcttgga atcaccatca
     7441 tggagcgttc ttctttcgag aagaacccaa tcgatttcct tgaggctaag ggatacaagg
     7501 aggttaagaa ggatcttatc atcaagttgc caaagtactc tcttttcgag cttgagaacg
     7561 gaagaaagag aatgcttgct tctgctggag agcttcagaa gggaaacgag cttgctcttc
     7621 catctaagta cgttaacttc ctttaccttg cttctcacta cgagaagttg aagggatctc
     7681 cagaggataa cgagcagaag cagcttttcg ttgagcagca caagcactac cttgatgaga
     7741 tcatcgagca aatctctgag ttctctaaga gagttatcct tgctgatgct aaccttgata
     7801 aggttctttc tgcttacaac aagcacagag ataagccaat cagagagcag gctgagaaca
     7861 tcatccacct tttcaccctt accaaccttg gtgctccagc tgctttcaag tacttcgata
     7921 ccaccatcga tagaaaaaga tacacctcta ccaaggaggt tcttgatgct acccttatcc
     7981 accagtctat caccggactt tacgagacca gaatcgatct ttctcagctt ggaggagata
     8041 agagaccagc tgctaccaag aaggctggac aggctaagaa gaagaagtga gacgtccgat
     8101 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     8161 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     8221 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     8281 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     8341 ttactagatc gggaattgat cccccctcga cagcttccgg aaagggcgaa ttcgcaactt
     8401 tgtatacaaa agttgccgag ctcgcggccg catgttgttg ttaccagaaa gtaaataaat
     8461 gttcaatctc tgatgttctc aagtaagtga gttttattgg gaataatatt aacttatgtt
     8521 cttcttgcat ttgatttctt tgccgctctc ttcttctatc ttaaatctgt gtatactatt
     8581 tcactattgg gctttttatt agtctataat gggactcaaa ataaggcttt ggcccacatc
     8641 aaaaagataa gtcacaaatc aaaactaaat tcagagtctt ttctcccaca tcggtcactg
     8701 tactcatttt gtgtttgttt atatattaca cgaaccgatc tttggtacga cagtccctac
     8761 atttgaagag ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     8821 tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg tacaaagttg
     8881 gcattaccat ggcttcgttg aacaacggaa actcgacttg ccttccgcac aatacatcat
     8941 ttcttcttag ctttttttct tcttcttcgt tcatacagtt tttttttgtt tatcagctta
     9001 cattttcttg aaccgtagct ttcgttttct tctttttaac tttccattcg gagtttttgt
     9061 atcttgtttc atagtttgtc ccaggattag aatgattagg catcgaacct tcaagaattt
     9121 gattgaataa aacatcttca ttcttaagat atgaagataa tcttcaaaag gcccctggga
     9181 atctgaaaga agagaagcag gcccatttat atgggaaaga acaatagtat ttcttatata
     9241 ggcccattta agttgaaaac aatcttcaaa agtcccacat cgcttagata agaaaacgaa
     9301 gctgagttta tatacagcta gagtcgaagt agtgattgat tagaacctaa aggaaccggt
     9361 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     9421 caccgagtcg gtgctttttt tctagaccca gctttcttgt acaaagttgg cattactgca
     9481 ggtcgacgac ccagctttct tgtacaaagt ggttcgataa ttcttaatta actagttcta
     9541 atctagagtc ctgctttaat gagatatgcg agacgcctat gatcgcatga tatttgcttt
     9601 caattctgtt gtgcacgttg taaaaaacct gagcatgtgt agctcagatc cttaccgccg
     9661 gtttcggttc attctaatga atatatcacc cgttactatc gtatttttat gaataatatt
     9721 ctccgttcaa tttactgatt gtaccctact acttatatgt acaatattaa aatgaaaaca
     9781 atatattgtg ctgaataggt ttatagcgac atctatgata gagcgccaca ataacaaaca
     9841 attgcgtttt attattacaa atccaatttt aaaaaaagcg gcagaaccgg tcaaacctaa
     9901 aagactgatt acataaatct tattcaaatt tcaaaagtgc cccaggggct agtatctacg
     9961 acacaccgag cggcgaacta ataacgctca ctgaagggaa ctccggttcc ccgccggcgc
    10021 gcatgggtga gattccttga agttgagtat tggccgtccg ctctaccgaa agttacgggc
    10081 accattcaac ccggtccagc acggcggccg ggtaaccgac ttgctgcccc gagaattatg
    10141 cagcattttt ttggtgtatg tgggccccaa atgaagtgca ggtcaaacct tgacagtgac
    10201 gacaaatcgt tgggcgggtc cagggcgaat tttgcgacaa catgtcgagg ctcagcagga
    10261 cctgcaggca tgcaagctag tgaagaatcg atttgctacg tattgcccct gcaggtcccc
    10321 agattagcct tttcaatttc agaaagaatg ctaacccaca gatggttaga gaggcttacg
    10381 cagcaggtct catcaagacg atctacccga gcaataatct ccaggaaatc aaataccttc
    10441 ccaagaaggt taaagatgca gtcaaaagat tcaggactaa ctgcatcaag aacacagaga
    10501 aagatatatt tctcaagatc agaagtacta ttccagtatg gacgattcaa ggcttgcttc
    10561 acaaaccaag gcaagtaata gagattggag tctctaaaaa ggtagttccc actgaatcaa
    10621 aggccatgga gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg
    10681 aacagttcat acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg
    10741 tggagcacga cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa
    10801 gggcaattga gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc
    10861 cagctatctg tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc
    10921 atcattgcga taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag
    10981 atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa
    11041 agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc
    11101 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagaga acacggggga
    11161 ctctagagga tccatgagcc cagaacgacg cccggccgac atccgccgtg ccaccgaggc
    11221 ggacatgccg gcggtctgca ccatcgtcaa ccactacatc gagacaagca cggtcaactt
    11281 ccgtaccgag ccgcaggaac cgcaggagtg gacggacgac ctcgtccgtc tgcgggagcg
    11341 ctatccctgg ctcgtcgccg aggtggacgg cgaggtcgcc ggcatcgcct acgcgggccc
    11401 ctggaaggca cgcaacgcct acgactggac ggccgagtcg accgtgtacg tctccccccg
    11461 ccaccagcgg acgggactgg gctccacgct ctacacccac ctgctgaagt ccctggaggc
    11521 acagggcttc aagagcgtgg tcgctgtcat cgggctgccc aacgacccga gcgtgcgcat
    11581 gcacgaggcg ctcggatatg ccccccgcgg catgctgcgg gcggccggct tcaagcacgg
    11641 gaactggcat gacgtgggtt tctggcagct ggacttcagc ctgccggtac cgccccgtcc
    11701 ggtcctgccc gtcaccgaga tctgaacgga gtgcgcgtgg catcgcccga gttggagctg
    11761 gtacgggaac tcatcgaact caactggcat acccgcaatg gtgaggtgga accgcggcgg
    11821 atcgcgtacg accgtgccca ggagggggta ccgagctcga atttccccga tcgttcaaac
    11881 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata
    11941 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt
    12001 atgagatggg tttttatgat tagagtcccg caattataca tttaatacgc gatagaaaac
    12061 aaaatatagc gcgcaaacta ggataaatta tcgcgcgcgg tgtcatctat gttactagat
    12121 cgggaattaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
    12181 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
    12241 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgcccgc tcctttcgct
    12301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
    12361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
    12421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
    12481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
    12541 tcgggctatt cttttgattt ataagggatt ttgccgattt cggaaccacc atcaaacagg
    12601 attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg
    12661 cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accccagtac
    12721 attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa
    12781 tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca
    12841 ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg
    12901 cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg
    12961 aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg
    13021 cctgtgatca aatatcatct ccctcgcaga gatccgaatt atcagccttc ttattcattt
    13081 ctcgcttaac cgtgacaggc tgtcgatctt gagaactatg ccgacataat aggaaatcgc
    13141 tggataaagc cgctgaggaa gctgagtggc gctatttctt tagaagtgaa cgttgacgat
    13201 atcaactccc ctatccattg ctcaccgaat ggtacaggtc ggggacccga agttccgact
    13261 gtcggcctga tgcatccccg gctgatcgac cccagatctg gggctgagaa agcccagtaa
    13321 ggaaacaact gtaggttcga gtcgcgagat cccccggaac caaaggaagt aggttaaacc
    13381 cgctccgatc aggccgagcc acgccaggcc gagaacattg gttcctgtag gcatcgggat
    13441 tggcggatca aacactaaag ctactggaac gagcagaagt cctccggccg ccagttgcca
    13501 ggcggtaaag gtgagcagag gcacgggagg ttgccacttg cgggtcagca cggttccgaa
    13561 cgccatggaa accgcccccg ccaggcccgc tgcgacgccg acaggatcta gcgctgcgtt
    13621 tggtgtcaac accaacagcg ccacgcccgc agttccgcaa atagccccca ggaccgccat
    13681 caatcgtatc gggctaccta gcagagcggc agagatgaac acgaccatca gcggctgcac
    13741 agcgcctacc gtcgccgcga ccccgcccgg caggcggtag accgaaataa acaacaagct
    13801 ccagaatagc gaaatattaa gtgcgccgag gatgaagatg cgcatccacc agattcccgt
    13861 tggaatctgt cggacgatca tcacgagcaa taaacccgcc ggcaacgccc gcagcagcat
    13921 accggcgacc cctcggcctc gctgttcggg ctccacgaaa acgccggaca gatgcgcctt
    13981 gtgagcgtcc ttggggccgt cctcctgttt gaagaccgac agcccaatga tctcgccgtc
    14041 gatgtaggcg ccgaatgcca cggcatctcg caaccgttca gcgaacgcct ccatgggctt
    14101 tttctcctcg tgctcgtaaa cggacccgaa catctctgga gctttcttca gggccgacaa
    14161 tcggatctcg cggaaatcct gcacgtcggc cgctccaagc cgtcgaatct gagccttaat
    14221 cacaattgtc aattttaatc ctctgtttat cggcagttcg tagagcgcgc cgtgcgtccc
    14281 gagcgatact gagcgaagca agtgcgtcga gcagtgcccg cttgttcctg aaatgccagt
    14341 aaagcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa
    14401 tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg
    14461 caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg
    14521 aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac
    14581 atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg
    14641 tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc
    14701 ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg
    14761 cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc
    14821 gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag
    14881 gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt
    14941 tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg
    15001 tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac
    15061 agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata
    15121 tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc
    15181 tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc
    15241 gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttcg
    15301 agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc
    15361 acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag
    15421 gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa
    15481 aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac
    15541 cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac
    15601 ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag
    15661 accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc
    15721 gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg
    15781 aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggtactaaaa
    15841 caattcatcc agtaaaatat aatattttat tttctcccaa tcaggcttga tccccagtaa
    15901 gtcaaaaaat agctcgacat actgttcttc cccgatatcc tccctgatcg accggacgca
    15961 gaaggcaatg tcataccact tgtccgccct gccgcttctc ccaagatcaa taaagccact
    16021 tactttgcca tctttcacaa agatgttgct gtctcccagg tcgccgtggg aaaagacaag
    16081 ttcctcttcg ggcttttccg tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg
    16141 agtgtcttct tcccagtttt cgcaatccac atcggccaga tcgttattca gtaagtaatc
    16201 caattcggct aagcggctgt ctaagctatt cgtataggga caatccgata tgtcgatgga
    16261 gtgaaagagc ctgatgcact ccgcatacag ctcgataatc ttttcagggc tttgttcatc
    16321 ttcatactct tccgagcaaa ggacgccatc ggcctcactc atgagcagat tgctccagcc
    16381 atcatgccgt tcaaagtgca ggacctttgg aacaggcagc tttccttcca gccatagcat
    16441 catgtccttt tcccgttcca catcataggt ggtcccttta taccggctgt ccgtcatttt
    16501 taaatatagg ttttcatttt ctcccaccag cttatatacc ttagcaggag acattccttc
    16561 cgtatctttt acgcagcggt atttttcgat cagttttttc aattccggtg atattctcat
    16621 tttagccatt tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct
    16681 aattataaca agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag
    16741 aaaacagctt tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg
    16801 attttgaaac cacaattatg ggtgatgctg ccaacttact gatttagtgt atgatggtgt
    16861 ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac
    16921 ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctatctctg ctctcactgc
    16981 cgtaaaacat ggcaactgca gttcacttac accgcttctc aacccggtac gcaccagaaa
    17041 atcattgata tggccatgaa tggcgttgga tgccgggcaa cagcccgcat tatgggcgtt
    17101 ggcctcaaca cgattttacg tcacttaaaa aactcaggcc gcagtcggta acctcgcgca
    17161 tacagccggg cagtgacgtc atcgtctgcg cggaaatgga cgaacagtgg ggctatgtcg
    17221 gggctaaatc gcgccagcgc tggctgtttt acgcgtatga cagtctccgg aagacggttg
    17281 ttgcgcacgt attcggtgaa cgcactatgg cgacgctggg gcgtcttatg agcctgctgt
    17341 caccctttga cgtggtgata tggatgacgg atggctggcc gctgtatgaa tcccgcctga
    17401 agggaaagct gcacgtaatc agcaagcgat atacgcagcg aattgagcgg cataacctga
    17461 atctgaggca gcacctggca cggctgggac ggaagtcgct gtcgttctca aaatcggtgg
    17521 agctgcatga caaagtcatc gggcattatc tgaacataaa acactatcaa taagttggag
    17581 tcattaccca attatgatag aatttacaag ctataaggtt attgtcctgg gtttcaagca
    17641 ttagtccatg caagttttta tgctttgccc attctataga tatattgata agcgcgctgc
    17701 ctatgccttg ccccctgaaa tccttacata cggcgatatc ttctatataa aagatatatt
    17761 atcttatcag tattgtcaat atattcaagg caatctgcct cctcatcctc ttcatcctct
    17821 tcgtcttggt agctttttaa atatggcgct tcatagagta attctgtaaa ggtccaattc
    17881 tcgttttcat acctcggtat aatcttacct atcacctcaa atggttcgct gggtttatcg
    17941 cacccccgaa cacgagcacg gcacccgcga ccactatgcc aagaatgccc aaggtaaaaa
    18001 ttgccggccc cgccatgaag tccgtgaatg ccccgacggc cgaagtgaag ggcaggccgc
    18061 cacccaggcc gccgccctca ctgcccggca cctggtcgct gaatgtcgat gccagcacct
    18121 gcggcacgtc aatgcttccg ggcgtcgcgc tcgggctgat cgcccatccc gttactgccc
    18181 cgatcccggc aatggcaagg actgccagcg ctgccatttt tggggtgagg ccgttcgcgg
    18241 ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg gagggttcga
    18301 gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta
    18361 aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg
    18421 aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat
    18481 gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc
    18541 cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg
    18601 cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag
    18661 gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg
    18721 ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag
    18781 ttttccgcga ggtatccaca acgccggcgg ccggccgcgg tgtctcgcac acggcttcga
    18841 cggcgtttct ggcgcgtttg cagggccata gacggccgcc agcccagcgg cgagggcaac
    18901 cagcccggtg agcgtcggaa aggcgctcgg tcttgccttg ctcgtcggtg atgtacttca
    18961 ccagctccgc gaagtcgctc ttcttgatg
//
