LOCUS       sequence-202769-       18988 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr4g094545 (Hen1). Also 
            expresses Cas9 from Gmubi promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18988)
  AUTHORS   Curtin SJ, Xiong Y, Michno JM, Campbell BW, Stec AO, Cermak T, 
            Starker C, Voytas DF, Eamens AL, Stupar RM
  TITLE     CRISPR/Cas9 and TALENs generate heritable mutations for genes 
            involved in small RNA processing of Glycine max and Medicago 
            truncatula.
  JOURNAL   Plant Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857.
  PUBMED    29087011
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"; journalName: "Plant 
            Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857."
COMMENT     Sequence Label: pSC29
FEATURES             Location/Qualifiers
     source          1..18988
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(98..469)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            502..611
                     /note="incP origin of transfer"
     primer_bind     834..853
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1087..1104
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1292..1942)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2407..2431
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3639..3663
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3685..3732
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3739..3759
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3784..5220,5410..8073)
                     /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          5221..5409
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8074..8121
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8132..8384
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8429..8449
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8804..8879
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8891..8915)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9392..9467
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9479..9503)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9522..9546)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9584..10291
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10841..11186
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11129..11149
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11207..11758
                     /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      11902..12154
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12167..12184)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12167..12183)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12176..12198)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12323..12342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12533..12554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12801..12825
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14421..15569)
                     /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(15868..16662)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16854..17621
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18244..18959
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cttcaccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt gcttggcaat
       61 cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc ctcgggcgga
      121 ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg
      181 cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca
      241 gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat
      301 agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag gccgacaggc
      361 tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca
      421 ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct
      481 catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg
      541 ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac
      601 ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc
      661 aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc
      721 cgaagcaggg ttatgcagcg gaaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      781 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      841 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      901 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      961 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1021 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1081 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc agaaggccgc cagagaggcc
     1141 gagcgcggcc gtgaggcttg gacgctaggg cagggcatga aaaagcccgt agcgggctgc
     1201 tacgggcgtc tgacgcggtg gaaaggggga ggggatgttg tctacatggc tctgctgtag
     1261 tgagtgggtt gcgctccggc agcggtcctg atcaatcgtc accctttctc ggtccttcaa
     1321 cgttcctgac aacgagcctc cttttcgcca atccatcgac aatcaccgcg agtccctgct
     1381 cgaacgctgc gtccggaccg gcttcgtcga aggcgtctat cgcggcccgc aacagcggcg
     1441 agagcggagc ctgttcaacg gtgccgccgc gctcgccggc atcgctgtcg ccggcctgct
     1501 cctcaagcac ggccccaaca gtgaagtagc tgattgtcat cagcgcattg acggcgtccc
     1561 cggccgaaaa acccgcctcg cagaggaagc gaagctgcgc gtcggccgtt tccatctgcg
     1621 gtgcgcccgg tcgcgtgccg gcatggatgc gcgcgccatc gcggtaggcg agcagcgcct
     1681 gcctgaagct gcgggcattc ccgatcagaa atgagcgcca gtcgtcgtcg gctctcggca
     1741 ccgaatgcgt atgattctcc gccagcatgg cttcggccag tgcgtcgagc agcgcccgct
     1801 tgttcctgaa gtgccagtaa agcgccggct gctgaacccc caaccgttcc gccagtttgc
     1861 gtgtcgtcag accgtctacg ccgacctcgt tcaacaggtc cagggcggca cggatcactg
     1921 tattcggctg caactttgtc atgcttgaca ctttatcact gataaacata atatgtccac
     1981 caacttatca gtgataaaga atccgcgcgt tcaatcggac cagcggaggc tggtccggag
     2041 gccagacgtg aaacccaaca tacccctgat cgtaattctg agcactgtcg cgctcgacgc
     2101 tgtcggcatc ggcctgatta tgccggtgct gccgggcctc ctgcgcgatc tggttcactc
     2161 gaacgacgtc accgcccact atggcattct gctggcgctg tatgcgttgg tgcaatttgc
     2221 ctgcgcacct gtgctgggcg cgctgtcgga tcgtttcggg cggcggccaa tcttgctcgt
     2281 ctcgctggcc ggcgccagat ctggggaacc ctgtggttgg catgcacata caaatggacg
     2341 aacggataaa ccttttcacg cccttttaaa tatccgatta ttctaataaa cgctcttttc
     2401 tcttaggttt acccgccaat atatcctgtc aaacactgat agtttaaact gaaggcggga
     2461 aacgacaatc tgatcatgag cggagaatta agggagtcac gttatgaccc ccgccgatga
     2521 cgcgggacaa gccgttttac gtttggaact gacagaaccg caacgttgaa ggagccactc
     2581 agccgcgggt ttctggagtt taatgagcta agcacatacg tcagaaacca ttattgcgcg
     2641 ttcaaaagtc gcctaaggtc actatcagct agacggacgc ggccgcgggc ccaatataac
     2701 aacgacgtcg taacagataa agcgaagctt gaaggtgcat gtgactccgt caagattacg
     2761 aaaccgccaa ctaccacgca aattgcaatt ctcaatttcc tagaaggact ctccgaaaat
     2821 gcatccaata ccaaatatta cccgtgtcat aggcaccaag tgacaccata catgaacacg
     2881 cgtcacaata tgactggaga agggttccac accttatgct ataaaacgcc ccacacccct
     2941 cctccttcct tcgcagttca attccaatat attccattct ctctgtgtat ttccctacct
     3001 ctcccttcaa ggttagtcga tttcttctgt ttttcttctt cgttctttcc atgaattgtg
     3061 tatgttcttt gatcaatacg atgttgattt gattgtgttt tgtttggttt catcgatctt
     3121 caattttcat aatcagattc agcttttatt atctttacaa caacgtcctt aatttgatga
     3181 ttctttaatc gtagatttgc tctaattaga gctttttcat gtcagatccc tttacaacaa
     3241 gccttaattg ttgattcatt aatcgtagat tagggctttt ttcattgatt acttcagatc
     3301 cgttaaacgt aaccatagat cagggctttt tcatgaatta cttcagatcc gttaaacaac
     3361 agccttattt tttatacttc tgtggttttt caagaaattg ttcagatccg ttgacaaaaa
     3421 gccttattcg ttgattctat atcgtttttc gagagatatt gctcagatct gttagcaact
     3481 gccttgtttg ttgattctat tgccgtggat tagggttttt tttcacgaga ttgcttcaga
     3541 tccgtactta agattacgta atggattttg attctgattt atctgtgatt gttgactcga
     3601 caggtacctt caaacctcga ggcgcgccaa gctatcaaac aagtttgtac aaaaaagcag
     3661 gctccgaatt cgcccttcac catggattac aaggatgatg atgataagga ttacaaggat
     3721 gatgatgata agatggctcc aaagaagaag agaaaggttg gaatccacgg agttccagct
     3781 gctgataaga agtactctat cggacttgac atcggaacca actctgttgg atgggctgtt
     3841 atcaccgatg agtacaaggt tccatctaag aagttcaagg ttcttggaaa caccgataga
     3901 cactctatca agaagaacct tatcggtgct cttcttttcg attctggaga gaccgctgag
     3961 gctaccagat tgaagagaac cgctagaaga agatacacca gaagaaagaa cagaatctgc
     4021 taccttcagg aaatcttctc taacgagatg gctaaggttg atgattcttt cttccacaga
     4081 cttgaggagt ctttccttgt tgaggaggat aagaagcacg agagacaccc aatcttcgga
     4141 aacatcgttg atgaggttgc ttaccacgag aagtacccaa ccatctacca ccttagaaag
     4201 aagttggttg attctaccga taaggctgat cttagactta tctaccttgc tcttgctcac
     4261 atgatcaagt tcagaggaca cttccttatc gagggagacc ttaacccaga taactctgat
     4321 gttgataagt tgttcatcca gcttgttcag acctacaacc agcttttcga ggagaaccca
     4381 atcaacgctt ctggagttga tgctaaggct atcctttctg ctagactttc taagtctcgt
     4441 agacttgaga accttatcgc tcagcttcca ggagagaaga agaacggact tttcggaaac
     4501 cttatcgctc tttctcttgg acttacccca aacttcaagt ctaacttcga tcttgctgag
     4561 gatgctaagt tgcagctttc taaggatacc tacgatgatg atcttgataa ccttcttgct
     4621 cagatcggag atcagtacgc tgatcttttc cttgctgcta agaacctttc tgatgctatc
     4681 cttctttctg acatccttag agttaacacc gagatcacca aggctccact ttctgcttct
     4741 atgatcaaga gatacgatga gcaccaccag gatcttaccc ttttgaaggc tcttgttaga
     4801 cagcagcttc cagagaagta caaggaaatc ttcttcgatc agtctaagaa cggatacgct
     4861 ggatacatcg atggaggagc ttctcaggag gagttctaca agttcatcaa gccaatcctt
     4921 gagaagatgg atggaaccga ggagcttctt gttaagttga acagagagga tcttcttaga
     4981 aagcagagaa ccttcgataa cggatctatc ccacaccaga tccaccttgg agagcttcac
     5041 gctatccttc gtagacagga ggatttctac ccattcttga aggataacag agagaagatc
     5101 gagaagatcc ttaccttcag aatcccatac tacgttggac cacttgctag aggaaactct
     5161 cgtttcgctt ggatgaccag aaagtctgag gagaccatca ccccttggaa cttcgaggag
     5221 gtaagtttct gcttctacct ttgatatata tataataatt atcattaatt agtagtaata
     5281 taatatttca aatatttttt tcaaaataaa agaatgtagt atatagcaat tgcttttctg
     5341 tagtttataa gtgtgtatat tttaatttat aacttttcta atatatgacc aaaatttgtt
     5401 gatgtgcagg ttgttgataa gggagcttct gctcagtctt tcatcgagag aatgaccaac
     5461 ttcgataaga accttccaaa cgagaaggtt cttccaaagc actctcttct ttacgagtac
     5521 ttcaccgttt acaacgagct taccaaggtt aagtacgtta ccgagggaat gagaaagcca
     5581 gctttccttt ctggagagca gaagaaggct atcgttgatc ttcttttcaa gaccaacaga
     5641 aaggttaccg ttaagcagtt gaaggaggat tacttcaaga agatcgagtg cttcgattct
     5701 gttgaaatct ctggagttga ggatagattc aacgcttctc ttggaaccta ccacgatctt
     5761 ttgaagatca tcaaggataa ggatttcctt gataacgagg agaacgagga catccttgag
     5821 gacatcgttc ttacccttac ccttttcgag gatagagaga tgatcgagga gagactcaag
     5881 acctacgctc accttttcga tgataaggtt atgaagcagt tgaagagaag aagatacacc
     5941 ggatggggta gactttctcg taagttgatc aacggaatca gagataagca gtctggaaag
     6001 accatccttg atttcttgaa gtctgatgga ttcgctaaca gaaacttcat gcagcttatc
     6061 cacgatgatt ctcttacctt caaggaggac atccagaagg ctcaggtttc tggacaggga
     6121 gattctcttc acgagcacat cgctaacctt gctggatctc cagctatcaa gaagggaatc
     6181 cttcagaccg ttaaggttgt tgatgagctt gttaaggtta tgggtagaca caagccagag
     6241 aacatcgtta tcgagatggc tagagagaac cagaccaccc agaagggaca gaagaactct
     6301 cgtgagagaa tgaagagaat cgaggaggga atcaaggagc ttggatctca aatcttgaag
     6361 gagcacccag ttgagaacac ccagcttcag aacgagaagt tgtaccttta ctaccttcag
     6421 aacggaagag atatgtacgt tgatcaggag cttgacatca acagactttc tgattacgat
     6481 gttgatcaca tcgttccaca gtctttcttg aaggatgatt ctatcgataa caaggttctt
     6541 acccgttctg ataagaacag aggaaagtct gataacgttc catctgagga ggttgttaag
     6601 aagatgaaga actactggag acagcttctt aacgctaagt tgatcaccca gagaaagttc
     6661 gataacctta ccaaggctga gagaggagga ctttctgagc ttgataaggc tggattcatc
     6721 aagagacagc ttgttgagac cagacagatc accaagcacg ttgctcagat ccttgattct
     6781 cgtatgaaca ccaagtacga tgagaacgat aagttgatca gagaggttaa ggttatcacc
     6841 ttgaagtcta agttggtttc tgatttcaga aaggatttcc agttctacaa ggttagagag
     6901 atcaacaact accaccacgc tcacgatgct taccttaacg ctgttgttgg aaccgctctt
     6961 atcaagaagt acccaaagtt ggagtctgag ttcgtttacg gagattacaa ggtttacgat
     7021 gttagaaaga tgatcgctaa gtctgagcag gagatcggaa aggctaccgc taagtacttc
     7081 ttctactcta acatcatgaa cttcttcaag accgagatca cccttgctaa cggagagatc
     7141 agaaagagac cacttatcga gaccaacgga gagaccggag agatcgtttg ggataaggga
     7201 agagatttcg ctaccgttag aaaggttctt tctatgccac aggttaacat cgttaagaaa
     7261 accgaggttc agaccggagg attctctaag gagtctatcc ttccaaagag aaactctgat
     7321 aagttgatcg ctagaaagaa ggattgggac ccaaagaagt acggaggatt cgattctcca
     7381 accgttgctt actctgttct tgttgttgct aaggttgaga agggaaagtc taagaagttg
     7441 aagtctgtta aggagcttct tggaatcacc atcatggagc gttcttcttt cgagaagaac
     7501 ccaatcgatt tccttgaggc taagggatac aaggaggtta agaaggatct tatcatcaag
     7561 ttgccaaagt actctctttt cgagcttgag aacggaagaa agagaatgct tgcttctgct
     7621 ggagagcttc agaagggaaa cgagcttgct cttccatcta agtacgttaa cttcctttac
     7681 cttgcttctc actacgagaa gttgaaggga tctccagagg ataacgagca gaagcagctt
     7741 ttcgttgagc agcacaagca ctaccttgat gagatcatcg agcaaatctc tgagttctct
     7801 aagagagtta tccttgctga tgctaacctt gataaggttc tttctgctta caacaagcac
     7861 agagataagc caatcagaga gcaggctgag aacatcatcc accttttcac ccttaccaac
     7921 cttggtgctc cagctgcttt caagtacttc gataccacca tcgatagaaa aagatacacc
     7981 tctaccaagg aggttcttga tgctaccctt atccaccagt ctatcaccgg actttacgag
     8041 accagaatcg atctttctca gcttggagga gataagagac cagctgctac caagaaggct
     8101 ggacaggcta agaagaagaa gtgagacgtc cgatcgttca aacatttggc aataaagttt
     8161 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
     8221 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
     8281 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
     8341 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat tgatcccccc
     8401 tcgacagctt ccggaaaggg cgaattcgca actttgtata caaaagttgc cgagctcgcg
     8461 gccgcatgtt gttgttacca gaaagtaaat aaatgttcaa tctctgatgt tctcaagtaa
     8521 gtgagtttta ttgggaataa tattaactta tgttcttctt gcatttgatt tctttgccgc
     8581 tctcttcttc tatcttaaat ctgtgtatac tatttcacta ttgggctttt tattagtcta
     8641 taatgggact caaaataagg ctttggccca catcaaaaag ataagtcaca aatcaaaact
     8701 aaattcagag tcttttctcc cacatcggtc actgtactca ttttgtgttt gtttatatat
     8761 tacacgaacc gatctttggt acgttacatc tgaacaacat atcgttttag agctagaaat
     8821 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     8881 ttttttctag acccagcttt cttgtacaaa gttggcatta ccatggcttc gttgaacaac
     8941 ggaaactcga cttgccttcc gcacaataca tcatttcttc ttagcttttt ttcttcttct
     9001 tcgttcatac agtttttttt tgtttatcag cttacatttt cttgaaccgt agctttcgtt
     9061 ttcttctttt taactttcca ttcggagttt ttgtatcttg tttcatagtt tgtcccagga
     9121 ttagaatgat taggcatcga accttcaaga atttgattga ataaaacatc ttcattctta
     9181 agatatgaag ataatcttca aaaggcccct gggaatctga aagaagagaa gcaggcccat
     9241 ttatatggga aagaacaata gtatttctta tataggccca tttaagttga aaacaatctt
     9301 caaaagtccc acatcgctta gataagaaaa cgaagctgag tttatataca gctagagtcg
     9361 aagtagtgat tgtccaaagc aatcagattc agttttagag ctagaaatag caagttaaaa
     9421 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttctagac
     9481 ccagctttct tgtacaaagt tggcattact gcaggtcgac gacccagctt tcttgtacaa
     9541 agtggttcga taattcttaa ttaactagtt ctaatctaga gtcctgcttt aatgagatat
     9601 gcgagacgcc tatgatcgca tgatatttgc tttcaattct gttgtgcacg ttgtaaaaaa
     9661 cctgagcatg tgtagctcag atccttaccg ccggtttcgg ttcattctaa tgaatatatc
     9721 acccgttact atcgtatttt tatgaataat attctccgtt caatttactg attgtaccct
     9781 actacttata tgtacaatat taaaatgaaa acaatatatt gtgctgaata ggtttatagc
     9841 gacatctatg atagagcgcc acaataacaa acaattgcgt tttattatta caaatccaat
     9901 tttaaaaaaa gcggcagaac cggtcaaacc taaaagactg attacataaa tcttattcaa
     9961 atttcaaaag tgccccaggg gctagtatct acgacacacc gagcggcgaa ctaataacgc
    10021 tcactgaagg gaactccggt tccccgccgg cgcgcatggg tgagattcct tgaagttgag
    10081 tattggccgt ccgctctacc gaaagttacg ggcaccattc aacccggtcc agcacggcgg
    10141 ccgggtaacc gacttgctgc cccgagaatt atgcagcatt tttttggtgt atgtgggccc
    10201 caaatgaagt gcaggtcaaa ccttgacagt gacgacaaat cgttgggcgg gtccagggcg
    10261 aattttgcga caacatgtcg aggctcagca ggacctgcag gcatgcaagc tagtgaagaa
    10321 tcgatttgct acgtattgcc cctgcaggtc cccagattag ccttttcaat ttcagaaaga
    10381 atgctaaccc acagatggtt agagaggctt acgcagcagg tctcatcaag acgatctacc
    10441 cgagcaataa tctccaggaa atcaaatacc ttcccaagaa ggttaaagat gcagtcaaaa
    10501 gattcaggac taactgcatc aagaacacag agaaagatat atttctcaag atcagaagta
    10561 ctattccagt atggacgatt caaggcttgc ttcacaaacc aaggcaagta atagagattg
    10621 gagtctctaa aaaggtagtt cccactgaat caaaggccat ggagtcaaag attcaaatag
    10681 aggacctaac agaactcgcc gtaaagactg gcgaacagtt catacagagt ctcttacgac
    10741 tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca cgacacactt gtctactcca
    10801 aaaatatcaa agatacagtc tcagaagacc aaagggcaat tgagactttt caacaaaggg
    10861 taatatccgg aaacctcctc ggattccatt gcccagctat ctgtcacttt attgtgaaga
    10921 tagtggaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggccatcg
    10981 ttgaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
    11041 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca
    11101 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
    11161 gaagttcatt tcatttggag agaacacggg ggactctaga ggatccatga gcccagaacg
    11221 acgcccggcc gacatccgcc gtgccaccga ggcggacatg ccggcggtct gcaccatcgt
    11281 caaccactac atcgagacaa gcacggtcaa cttccgtacc gagccgcagg aaccgcagga
    11341 gtggacggac gacctcgtcc gtctgcggga gcgctatccc tggctcgtcg ccgaggtgga
    11401 cggcgaggtc gccggcatcg cctacgcggg cccctggaag gcacgcaacg cctacgactg
    11461 gacggccgag tcgaccgtgt acgtctcccc ccgccaccag cggacgggac tgggctccac
    11521 gctctacacc cacctgctga agtccctgga ggcacagggc ttcaagagcg tggtcgctgt
    11581 catcgggctg cccaacgacc cgagcgtgcg catgcacgag gcgctcggat atgccccccg
    11641 cggcatgctg cgggcggccg gcttcaagca cgggaactgg catgacgtgg gtttctggca
    11701 gctggacttc agcctgccgg taccgccccg tccggtcctg cccgtcaccg agatctgaac
    11761 ggagtgcgcg tggcatcgcc cgagttggag ctggtacggg aactcatcga actcaactgg
    11821 catacccgca atggtgaggt ggaaccgcgg cggatcgcgt acgaccgtgc ccaggagggg
    11881 gtaccgagct cgaatttccc cgatcgttca aacatttggc aataaagttt cttaagattg
    11941 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat
    12001 gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc
    12061 ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa
    12121 ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat taattcactg gccgtcgttt
    12181 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
    12241 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
    12301 tgcgcagcct gaatggcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    12361 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    12421 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
    12481 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    12541 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
    12601 attttgccga tttcggaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
    12661 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
    12721 tctcactggt gaaaagaaaa accaccccag tacattaaaa acgtccgcaa tgtgttatta
    12781 agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc cagccaacag
    12841 ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc atcagtccgg
    12901 gacggcgtca gcgggagagc cgttgtaagg cggcagactt tgctcatgtt accgatgcta
    12961 ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag
    13021 catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaaatatca tctccctcgc
    13081 agagatccga attatcagcc ttcttattca tttctcgctt aaccgtgaca ggctgtcgat
    13141 cttgagaact atgccgacat aataggaaat cgctggataa agccgctgag gaagctgagt
    13201 ggcgctattt ctttagaagt gaacgttgac gatatcaact cccctatcca ttgctcaccg
    13261 aatggtacag gtcggggacc cgaagttccg actgtcggcc tgatgcatcc ccggctgatc
    13321 gaccccagat ctggggctga gaaagcccag taaggaaaca actgtaggtt cgagtcgcga
    13381 gatcccccgg aaccaaagga agtaggttaa acccgctccg atcaggccga gccacgccag
    13441 gccgagaaca ttggttcctg taggcatcgg gattggcgga tcaaacacta aagctactgg
    13501 aacgagcaga agtcctccgg ccgccagttg ccaggcggta aaggtgagca gaggcacggg
    13561 aggttgccac ttgcgggtca gcacggttcc gaacgccatg gaaaccgccc ccgccaggcc
    13621 cgctgcgacg ccgacaggat ctagcgctgc gtttggtgtc aacaccaaca gcgccacgcc
    13681 cgcagttccg caaatagccc ccaggaccgc catcaatcgt atcgggctac ctagcagagc
    13741 ggcagagatg aacacgacca tcagcggctg cacagcgcct accgtcgccg cgaccccgcc
    13801 cggcaggcgg tagaccgaaa taaacaacaa gctccagaat agcgaaatat taagtgcgcc
    13861 gaggatgaag atgcgcatcc accagattcc cgttggaatc tgtcggacga tcatcacgag
    13921 caataaaccc gccggcaacg cccgcagcag cataccggcg acccctcggc ctcgctgttc
    13981 gggctccacg aaaacgccgg acagatgcgc cttgtgagcg tccttggggc cgtcctcctg
    14041 tttgaagacc gacagcccaa tgatctcgcc gtcgatgtag gcgccgaatg ccacggcatc
    14101 tcgcaaccgt tcagcgaacg cctccatggg ctttttctcc tcgtgctcgt aaacggaccc
    14161 gaacatctct ggagctttct tcagggccga caatcggatc tcgcggaaat cctgcacgtc
    14221 ggccgctcca agccgtcgaa tctgagcctt aatcacaatt gtcaatttta atcctctgtt
    14281 tatcggcagt tcgtagagcg cgccgtgcgt cccgagcgat actgagcgaa gcaagtgcgt
    14341 cgagcagtgc ccgcttgttc ctgaaatgcc agtaaagcgc tggctgctga acccccagcc
    14401 ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg
    14461 tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac
    14521 ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg
    14581 tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg
    14641 cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc
    14701 tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg
    14761 tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc
    14821 gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag
    14881 cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc
    14941 ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg
    15001 ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc
    15061 tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc
    15121 atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg
    15181 atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc
    15241 aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc
    15301 atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc
    15361 gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta
    15421 gcttgctgga ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg
    15481 cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg
    15541 tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccgactcac
    15601 gccggggcaa tgtgccctta ttcctgattt gacccgcctg gtgccttggt gtccagataa
    15661 tccaccttat cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg
    15721 gtattccgaa tcttgccctg cacgaatacc agcgacccct tgcccaaata cttgccgtgg
    15781 gcctcggcct gagagccaaa acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg
    15841 ccggcatcgt tgcgccacat ctaggtacta aaacaattca tccagtaaaa tataatattt
    15901 tattttctcc caatcaggct tgatccccag taagtcaaaa aatagctcga catactgttc
    15961 ttccccgata tcctccctga tcgaccggac gcagaaggca atgtcatacc acttgtccgc
    16021 cctgccgctt ctcccaagat caataaagcc acttactttg ccatctttca caaagatgtt
    16081 gctgtctccc aggtcgccgt gggaaaagac aagttcctct tcgggctttt ccgtctttaa
    16141 aaaatcatac agctcgcgcg gatctttaaa tggagtgtct tcttcccagt tttcgcaatc
    16201 cacatcggcc agatcgttat tcagtaagta atccaattcg gctaagcggc tgtctaagct
    16261 attcgtatag ggacaatccg atatgtcgat ggagtgaaag agcctgatgc actccgcata
    16321 cagctcgata atcttttcag ggctttgttc atcttcatac tcttccgagc aaaggacgcc
    16381 atcggcctca ctcatgagca gattgctcca gccatcatgc cgttcaaagt gcaggacctt
    16441 tggaacaggc agctttcctt ccagccatag catcatgtcc ttttcccgtt ccacatcata
    16501 ggtggtccct ttataccggc tgtccgtcat ttttaaatat aggttttcat tttctcccac
    16561 cagcttatat accttagcag gagacattcc ttccgtatct tttacgcagc ggtatttttc
    16621 gatcagtttt ttcaattccg gtgatattct cattttagcc atttattatt tccttcctct
    16681 tttctacagt atttaaagat accccaagaa gctaattata acaagacgaa ctccaattca
    16741 ctgttccttg cattctaaaa ccttaaatac cagaaaacag ctttttcaaa gttgttttca
    16801 aagttggcgt ataacatagt atcgacggag ccgattttga aaccacaatt atgggtgatg
    16861 ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt
    16921 tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc
    16981 gccggacatc agcgctatct ctgctctcac tgccgtaaaa catggcaact gcagttcact
    17041 tacaccgctt ctcaacccgg tacgcaccag aaaatcattg atatggccat gaatggcgtt
    17101 ggatgccggg caacagcccg cattatgggc gttggcctca acacgatttt acgtcactta
    17161 aaaaactcag gccgcagtcg gtaacctcgc gcatacagcc gggcagtgac gtcatcgtct
    17221 gcgcggaaat ggacgaacag tggggctatg tcggggctaa atcgcgccag cgctggctgt
    17281 tttacgcgta tgacagtctc cggaagacgg ttgttgcgca cgtattcggt gaacgcacta
    17341 tggcgacgct ggggcgtctt atgagcctgc tgtcaccctt tgacgtggtg atatggatga
    17401 cggatggctg gccgctgtat gaatcccgcc tgaagggaaa gctgcacgta atcagcaagc
    17461 gatatacgca gcgaattgag cggcataacc tgaatctgag gcagcacctg gcacggctgg
    17521 gacggaagtc gctgtcgttc tcaaaatcgg tggagctgca tgacaaagtc atcgggcatt
    17581 atctgaacat aaaacactat caataagttg gagtcattac ccaattatga tagaatttac
    17641 aagctataag gttattgtcc tgggtttcaa gcattagtcc atgcaagttt ttatgctttg
    17701 cccattctat agatatattg ataagcgcgc tgcctatgcc ttgccccctg aaatccttac
    17761 atacggcgat atcttctata taaaagatat attatcttat cagtattgtc aatatattca
    17821 aggcaatctg cctcctcatc ctcttcatcc tcttcgtctt ggtagctttt taaatatggc
    17881 gcttcataga gtaattctgt aaaggtccaa ttctcgtttt catacctcgg tataatctta
    17941 cctatcacct caaatggttc gctgggttta tcgcaccccc gaacacgagc acggcacccg
    18001 cgaccactat gccaagaatg cccaaggtaa aaattgccgg ccccgccatg aagtccgtga
    18061 atgccccgac ggccgaagtg aagggcaggc cgccacccag gccgccgccc tcactgcccg
    18121 gcacctggtc gctgaatgtc gatgccagca cctgcggcac gtcaatgctt ccgggcgtcg
    18181 cgctcgggct gatcgcccat cccgttactg ccccgatccc ggcaatggca aggactgcca
    18241 gcgctgccat ttttggggtg aggccgttcg cggccgaggg gcgcagcccc tggggggatg
    18301 ggaggcccgc gttagcgggc cgggagggtt cgagaagggg gggcaccccc cttcggcgtg
    18361 cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa ggtttataaa tattggttta
    18421 aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg ggcggaaacc cttgcaaatg
    18481 ctggattttc tgcctgtgga cagcccctca aatgtcaata ggtgcgcccc tcatctgtca
    18541 gcactctgcc cctcaagtgt caaggatcgc gcccctcatc tgtcagtagt cgcgcccctc
    18601 aagtgtcaat accgcagggc acttatcccc aggcttgtcc acatcatctg tgggaaactc
    18661 gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc agctccacgt cgccggccga
    18721 aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg agtcggcccc tcaagtgtca
    18781 acgtccgccc ctcatctgtc agtgagggcc aagttttccg cgaggtatcc acaacgccgg
    18841 cggccggccg cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt ttgcagggcc
    18901 atagacggcc gccagcccag cggcgagggc aaccagcccg gtgagcgtcg gaaaggcgct
    18961 cggtcttgcc ttgctcgtcg gtgatgta
//
