LOCUS       sequence-203137-       18989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Glyma.08g081600, Glyma.05g126600 
            (Hen1ab). 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, 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 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"; journalName: "Plant 
            Biotechnol J. 2017 Oct 31. doi: 10.1111/pbi.12857."
COMMENT     Sequence Label: pSC50
FEATURES             Location/Qualifiers
     source          1..18989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(103..474)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            507..616
                     /note="incP origin of transfer"
     primer_bind     839..858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1092..1109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1297..1947)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2412..2436
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3644..3668
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3690..3737
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3744..3764
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3789..5225,5415..8078)
                     /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          5226..5414
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8079..8126
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8137..8389
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8434..8454
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8809..8884
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8896..8920)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9398..9473
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9485..9509)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9528..9552)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9590..10297
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10847..11192
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11135..11155
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11213..11764
                     /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      11908..12160
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12173..12190)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12173..12189)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12182..12204)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12329..12348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12539..12560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12807..12831
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14427..15575)
                     /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(15874..16668)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16860..17627
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18250..18965
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 atgtacttca ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg
       61 gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct ctgccctcgg
      121 gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag
      181 cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag
      241 tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct cgcggacgtg
      301 ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca ggtaggccga
      361 caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca
      421 gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
      481 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
      541 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
      601 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
      661 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
      721 gaccccgaag cagggttatg cagcggaaaa gcgccacgct tcccgaaggg agaaaggcgg
      781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
      841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
      901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     1021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     1081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgccagaag gccgccagag
     1141 aggccgagcg cggccgtgag gcttggacgc tagggcaggg catgaaaaag cccgtagcgg
     1201 gctgctacgg gcgtctgacg cggtggaaag ggggagggga tgttgtctac atggctctgc
     1261 tgtagtgagt gggttgcgct ccggcagcgg tcctgatcaa tcgtcaccct ttctcggtcc
     1321 ttcaacgttc ctgacaacga gcctcctttt cgccaatcca tcgacaatca ccgcgagtcc
     1381 ctgctcgaac gctgcgtccg gaccggcttc gtcgaaggcg tctatcgcgg cccgcaacag
     1441 cggcgagagc ggagcctgtt caacggtgcc gccgcgctcg ccggcatcgc tgtcgccggc
     1501 ctgctcctca agcacggccc caacagtgaa gtagctgatt gtcatcagcg cattgacggc
     1561 gtccccggcc gaaaaacccg cctcgcagag gaagcgaagc tgcgcgtcgg ccgtttccat
     1621 ctgcggtgcg cccggtcgcg tgccggcatg gatgcgcgcg ccatcgcggt aggcgagcag
     1681 cgcctgcctg aagctgcggg cattcccgat cagaaatgag cgccagtcgt cgtcggctct
     1741 cggcaccgaa tgcgtatgat tctccgccag catggcttcg gccagtgcgt cgagcagcgc
     1801 ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga acccccaacc gttccgccag
     1861 tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac aggtccaggg cggcacggat
     1921 cactgtattc ggctgcaact ttgtcatgct tgacacttta tcactgataa acataatatg
     1981 tccaccaact tatcagtgat aaagaatccg cgcgttcaat cggaccagcg gaggctggtc
     2041 cggaggccag acgtgaaacc caacataccc ctgatcgtaa ttctgagcac tgtcgcgctc
     2101 gacgctgtcg gcatcggcct gattatgccg gtgctgccgg gcctcctgcg cgatctggtt
     2161 cactcgaacg acgtcaccgc ccactatggc attctgctgg cgctgtatgc gttggtgcaa
     2221 tttgcctgcg cacctgtgct gggcgcgctg tcggatcgtt tcgggcggcg gccaatcttg
     2281 ctcgtctcgc tggccggcgc cagatctggg gaaccctgtg gttggcatgc acatacaaat
     2341 ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta ataaacgctc
     2401 ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aaactgaagg
     2461 cgggaaacga caatctgatc atgagcggag aattaaggga gtcacgttat gacccccgcc
     2521 gatgacgcgg gacaagccgt tttacgtttg gaactgacag aaccgcaacg ttgaaggagc
     2581 cactcagccg cgggtttctg gagtttaatg agctaagcac atacgtcaga aaccattatt
     2641 gcgcgttcaa aagtcgccta aggtcactat cagctagacg gacgcggccg cgggcccaat
     2701 ataacaacga cgtcgtaaca gataaagcga agcttgaagg tgcatgtgac tccgtcaaga
     2761 ttacgaaacc gccaactacc acgcaaattg caattctcaa tttcctagaa ggactctccg
     2821 aaaatgcatc caataccaaa tattacccgt gtcataggca ccaagtgaca ccatacatga
     2881 acacgcgtca caatatgact ggagaagggt tccacacctt atgctataaa acgccccaca
     2941 cccctcctcc ttccttcgca gttcaattcc aatatattcc attctctctg tgtatttccc
     3001 tacctctccc ttcaaggtta gtcgatttct tctgtttttc ttcttcgttc tttccatgaa
     3061 ttgtgtatgt tctttgatca atacgatgtt gatttgattg tgttttgttt ggtttcatcg
     3121 atcttcaatt ttcataatca gattcagctt ttattatctt tacaacaacg tccttaattt
     3181 gatgattctt taatcgtaga tttgctctaa ttagagcttt ttcatgtcag atccctttac
     3241 aacaagcctt aattgttgat tcattaatcg tagattaggg cttttttcat tgattacttc
     3301 agatccgtta aacgtaacca tagatcaggg ctttttcatg aattacttca gatccgttaa
     3361 acaacagcct tattttttat acttctgtgg tttttcaaga aattgttcag atccgttgac
     3421 aaaaagcctt attcgttgat tctatatcgt ttttcgagag atattgctca gatctgttag
     3481 caactgcctt gtttgttgat tctattgccg tggattaggg ttttttttca cgagattgct
     3541 tcagatccgt acttaagatt acgtaatgga ttttgattct gatttatctg tgattgttga
     3601 ctcgacaggt accttcaaac ctcgaggcgc gccaagctat caaacaagtt tgtacaaaaa
     3661 agcaggctcc gaattcgccc ttcaccatgg attacaagga tgatgatgat aaggattaca
     3721 aggatgatga tgataagatg gctccaaaga agaagagaaa ggttggaatc cacggagttc
     3781 cagctgctga taagaagtac tctatcggac ttgacatcgg aaccaactct gttggatggg
     3841 ctgttatcac cgatgagtac aaggttccat ctaagaagtt caaggttctt ggaaacaccg
     3901 atagacactc tatcaagaag aaccttatcg gtgctcttct tttcgattct ggagagaccg
     3961 ctgaggctac cagattgaag agaaccgcta gaagaagata caccagaaga aagaacagaa
     4021 tctgctacct tcaggaaatc ttctctaacg agatggctaa ggttgatgat tctttcttcc
     4081 acagacttga ggagtctttc cttgttgagg aggataagaa gcacgagaga cacccaatct
     4141 tcggaaacat cgttgatgag gttgcttacc acgagaagta cccaaccatc taccacctta
     4201 gaaagaagtt ggttgattct accgataagg ctgatcttag acttatctac cttgctcttg
     4261 ctcacatgat caagttcaga ggacacttcc ttatcgaggg agaccttaac ccagataact
     4321 ctgatgttga taagttgttc atccagcttg ttcagaccta caaccagctt ttcgaggaga
     4381 acccaatcaa cgcttctgga gttgatgcta aggctatcct ttctgctaga ctttctaagt
     4441 ctcgtagact tgagaacctt atcgctcagc ttccaggaga gaagaagaac ggacttttcg
     4501 gaaaccttat cgctctttct cttggactta ccccaaactt caagtctaac ttcgatcttg
     4561 ctgaggatgc taagttgcag ctttctaagg atacctacga tgatgatctt gataaccttc
     4621 ttgctcagat cggagatcag tacgctgatc ttttccttgc tgctaagaac ctttctgatg
     4681 ctatccttct ttctgacatc cttagagtta acaccgagat caccaaggct ccactttctg
     4741 cttctatgat caagagatac gatgagcacc accaggatct tacccttttg aaggctcttg
     4801 ttagacagca gcttccagag aagtacaagg aaatcttctt cgatcagtct aagaacggat
     4861 acgctggata catcgatgga ggagcttctc aggaggagtt ctacaagttc atcaagccaa
     4921 tccttgagaa gatggatgga accgaggagc ttcttgttaa gttgaacaga gaggatcttc
     4981 ttagaaagca gagaaccttc gataacggat ctatcccaca ccagatccac cttggagagc
     5041 ttcacgctat ccttcgtaga caggaggatt tctacccatt cttgaaggat aacagagaga
     5101 agatcgagaa gatccttacc ttcagaatcc catactacgt tggaccactt gctagaggaa
     5161 actctcgttt cgcttggatg accagaaagt ctgaggagac catcacccct tggaacttcg
     5221 aggaggtaag tttctgcttc tacctttgat atatatataa taattatcat taattagtag
     5281 taatataata tttcaaatat ttttttcaaa ataaaagaat gtagtatata gcaattgctt
     5341 ttctgtagtt tataagtgtg tatattttaa tttataactt ttctaatata tgaccaaaat
     5401 ttgttgatgt gcaggttgtt gataagggag cttctgctca gtctttcatc gagagaatga
     5461 ccaacttcga taagaacctt ccaaacgaga aggttcttcc aaagcactct cttctttacg
     5521 agtacttcac cgtttacaac gagcttacca aggttaagta cgttaccgag ggaatgagaa
     5581 agccagcttt cctttctgga gagcagaaga aggctatcgt tgatcttctt ttcaagacca
     5641 acagaaaggt taccgttaag cagttgaagg aggattactt caagaagatc gagtgcttcg
     5701 attctgttga aatctctgga gttgaggata gattcaacgc ttctcttgga acctaccacg
     5761 atcttttgaa gatcatcaag gataaggatt tccttgataa cgaggagaac gaggacatcc
     5821 ttgaggacat cgttcttacc cttacccttt tcgaggatag agagatgatc gaggagagac
     5881 tcaagaccta cgctcacctt ttcgatgata aggttatgaa gcagttgaag agaagaagat
     5941 acaccggatg gggtagactt tctcgtaagt tgatcaacgg aatcagagat aagcagtctg
     6001 gaaagaccat ccttgatttc ttgaagtctg atggattcgc taacagaaac ttcatgcagc
     6061 ttatccacga tgattctctt accttcaagg aggacatcca gaaggctcag gtttctggac
     6121 agggagattc tcttcacgag cacatcgcta accttgctgg atctccagct atcaagaagg
     6181 gaatccttca gaccgttaag gttgttgatg agcttgttaa ggttatgggt agacacaagc
     6241 cagagaacat cgttatcgag atggctagag agaaccagac cacccagaag ggacagaaga
     6301 actctcgtga gagaatgaag agaatcgagg agggaatcaa ggagcttgga tctcaaatct
     6361 tgaaggagca cccagttgag aacacccagc ttcagaacga gaagttgtac ctttactacc
     6421 ttcagaacgg aagagatatg tacgttgatc aggagcttga catcaacaga ctttctgatt
     6481 acgatgttga tcacatcgtt ccacagtctt tcttgaagga tgattctatc gataacaagg
     6541 ttcttacccg ttctgataag aacagaggaa agtctgataa cgttccatct gaggaggttg
     6601 ttaagaagat gaagaactac tggagacagc ttcttaacgc taagttgatc acccagagaa
     6661 agttcgataa ccttaccaag gctgagagag gaggactttc tgagcttgat aaggctggat
     6721 tcatcaagag acagcttgtt gagaccagac agatcaccaa gcacgttgct cagatccttg
     6781 attctcgtat gaacaccaag tacgatgaga acgataagtt gatcagagag gttaaggtta
     6841 tcaccttgaa gtctaagttg gtttctgatt tcagaaagga tttccagttc tacaaggtta
     6901 gagagatcaa caactaccac cacgctcacg atgcttacct taacgctgtt gttggaaccg
     6961 ctcttatcaa gaagtaccca aagttggagt ctgagttcgt ttacggagat tacaaggttt
     7021 acgatgttag aaagatgatc gctaagtctg agcaggagat cggaaaggct accgctaagt
     7081 acttcttcta ctctaacatc atgaacttct tcaagaccga gatcaccctt gctaacggag
     7141 agatcagaaa gagaccactt atcgagacca acggagagac cggagagatc gtttgggata
     7201 agggaagaga tttcgctacc gttagaaagg ttctttctat gccacaggtt aacatcgtta
     7261 agaaaaccga ggttcagacc ggaggattct ctaaggagtc tatccttcca aagagaaact
     7321 ctgataagtt gatcgctaga aagaaggatt gggacccaaa gaagtacgga ggattcgatt
     7381 ctccaaccgt tgcttactct gttcttgttg ttgctaaggt tgagaaggga aagtctaaga
     7441 agttgaagtc tgttaaggag cttcttggaa tcaccatcat ggagcgttct tctttcgaga
     7501 agaacccaat cgatttcctt gaggctaagg gatacaagga ggttaagaag gatcttatca
     7561 tcaagttgcc aaagtactct cttttcgagc ttgagaacgg aagaaagaga atgcttgctt
     7621 ctgctggaga gcttcagaag ggaaacgagc ttgctcttcc atctaagtac gttaacttcc
     7681 tttaccttgc ttctcactac gagaagttga agggatctcc agaggataac gagcagaagc
     7741 agcttttcgt tgagcagcac aagcactacc ttgatgagat catcgagcaa atctctgagt
     7801 tctctaagag agttatcctt gctgatgcta accttgataa ggttctttct gcttacaaca
     7861 agcacagaga taagccaatc agagagcagg ctgagaacat catccacctt ttcaccctta
     7921 ccaaccttgg tgctccagct gctttcaagt acttcgatac caccatcgat agaaaaagat
     7981 acacctctac caaggaggtt cttgatgcta cccttatcca ccagtctatc accggacttt
     8041 acgagaccag aatcgatctt tctcagcttg gaggagataa gagaccagct gctaccaaga
     8101 aggctggaca ggctaagaag aagaagtgag acgtccgatc gttcaaacat ttggcaataa
     8161 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
     8221 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
     8281 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
     8341 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattgatc
     8401 ccccctcgac agcttccgga aagggcgaat tcgcaacttt gtatacaaaa gttgccgagc
     8461 tcgcggccgc atgttgttgt taccagaaag taaataaatg ttcaatctct gatgttctca
     8521 agtaagtgag ttttattggg aataatatta acttatgttc ttcttgcatt tgatttcttt
     8581 gccgctctct tcttctatct taaatctgtg tatactattt cactattggg ctttttatta
     8641 gtctataatg ggactcaaaa taaggctttg gcccacatca aaaagataag tcacaaatca
     8701 aaactaaatt cagagtcttt tctcccacat cggtcactgt actcattttg tgtttgttta
     8761 tatattacac gaaccgatct ttggtacgaa agggatctct gaatgtaagt tttagagcta
     8821 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     8881 gtgctttttt tctagaccca gctttcttgt acaaagttgg cattaccatg gcttcgttga
     8941 acaacggaaa ctcgacttgc cttccgcaca atacatcatt tcttcttagc tttttttctt
     9001 cttcttcgtt catacagttt ttttttgttt atcagcttac attttcttga accgtagctt
     9061 tcgttttctt ctttttaact ttccattcgg agtttttgta tcttgtttca tagtttgtcc
     9121 caggattaga atgattaggc atcgaacctt caagaatttg attgaataaa acatcttcat
     9181 tcttaagata tgaagataat cttcaaaagg cccctgggaa tctgaaagaa gagaagcagg
     9241 cccatttata tgggaaagaa caatagtatt tcttatatag gcccatttaa gttgaaaaca
     9301 atcttcaaaa gtcccacatc gcttagataa gaaaacgaag ctgagtttat atacagctag
     9361 agtcgaagta gtgattgagt ttcttgctac ttctgaagtt ttagagctag aaatagcaag
     9421 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     9481 ctagacccag ctttcttgta caaagttggc attactgcag gtcgacgacc cagctttctt
     9541 gtacaaagtg gttcgataat tcttaattaa ctagttctaa tctagagtcc tgctttaatg
     9601 agatatgcga gacgcctatg atcgcatgat atttgctttc aattctgttg tgcacgttgt
     9661 aaaaaacctg agcatgtgta gctcagatcc ttaccgccgg tttcggttca ttctaatgaa
     9721 tatatcaccc gttactatcg tatttttatg aataatattc tccgttcaat ttactgattg
     9781 taccctacta cttatatgta caatattaaa atgaaaacaa tatattgtgc tgaataggtt
     9841 tatagcgaca tctatgatag agcgccacaa taacaaacaa ttgcgtttta ttattacaaa
     9901 tccaatttta aaaaaagcgg cagaaccggt caaacctaaa agactgatta cataaatctt
     9961 attcaaattt caaaagtgcc ccaggggcta gtatctacga cacaccgagc ggcgaactaa
    10021 taacgctcac tgaagggaac tccggttccc cgccggcgcg catgggtgag attccttgaa
    10081 gttgagtatt ggccgtccgc tctaccgaaa gttacgggca ccattcaacc cggtccagca
    10141 cggcggccgg gtaaccgact tgctgccccg agaattatgc agcatttttt tggtgtatgt
    10201 gggccccaaa tgaagtgcag gtcaaacctt gacagtgacg acaaatcgtt gggcgggtcc
    10261 agggcgaatt ttgcgacaac atgtcgaggc tcagcaggac ctgcaggcat gcaagctagt
    10321 gaagaatcga tttgctacgt attgcccctg caggtcccca gattagcctt ttcaatttca
    10381 gaaagaatgc taacccacag atggttagag aggcttacgc agcaggtctc atcaagacga
    10441 tctacccgag caataatctc caggaaatca aataccttcc caagaaggtt aaagatgcag
    10501 tcaaaagatt caggactaac tgcatcaaga acacagagaa agatatattt ctcaagatca
    10561 gaagtactat tccagtatgg acgattcaag gcttgcttca caaaccaagg caagtaatag
    10621 agattggagt ctctaaaaag gtagttccca ctgaatcaaa ggccatggag tcaaagattc
    10681 aaatagagga cctaacagaa ctcgccgtaa agactggcga acagttcata cagagtctct
    10741 tacgactcaa tgacaagaag aaaatcttcg tcaacatggt ggagcacgac acacttgtct
    10801 actccaaaaa tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac
    10861 aaagggtaat atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg
    10921 tgaagatagt ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg
    10981 ccatcgttga agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga
    11041 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata
    11101 tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta
    11161 tataaggaag ttcatttcat ttggagagaa cacgggggac tctagaggat ccatgagccc
    11221 agaacgacgc ccggccgaca tccgccgtgc caccgaggcg gacatgccgg cggtctgcac
    11281 catcgtcaac cactacatcg agacaagcac ggtcaacttc cgtaccgagc cgcaggaacc
    11341 gcaggagtgg acggacgacc tcgtccgtct gcgggagcgc tatccctggc tcgtcgccga
    11401 ggtggacggc gaggtcgccg gcatcgccta cgcgggcccc tggaaggcac gcaacgccta
    11461 cgactggacg gccgagtcga ccgtgtacgt ctccccccgc caccagcgga cgggactggg
    11521 ctccacgctc tacacccacc tgctgaagtc cctggaggca cagggcttca agagcgtggt
    11581 cgctgtcatc gggctgccca acgacccgag cgtgcgcatg cacgaggcgc tcggatatgc
    11641 cccccgcggc atgctgcggg cggccggctt caagcacggg aactggcatg acgtgggttt
    11701 ctggcagctg gacttcagcc tgccggtacc gccccgtccg gtcctgcccg tcaccgagat
    11761 ctgaacggag tgcgcgtggc atcgcccgag ttggagctgg tacgggaact catcgaactc
    11821 aactggcata cccgcaatgg tgaggtggaa ccgcggcgga tcgcgtacga ccgtgcccag
    11881 gagggggtac cgagctcgaa tttccccgat cgttcaaaca tttggcaata aagtttctta
    11941 agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt
    12001 aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt
    12061 agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag
    12121 gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattaat tcactggccg
    12181 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
    12241 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
    12301 aacagttgcg cagcctgaat ggcgcccgct cctttcgctt tcttcccttc ctttctcgcc
    12361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
    12421 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
    12481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
    12541 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
    12601 taagggattt tgccgatttc ggaaccacca tcaaacagga ttttcgcctg ctggggcaaa
    12661 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
    12721 tgcccgtctc actggtgaaa agaaaaacca ccccagtaca ttaaaaacgt ccgcaatgtg
    12781 ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc
    12841 caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca
    12901 gtccgggacg gcgtcagcgg gagagccgtt gtaaggcggc agactttgct catgttaccg
    12961 atgctattcg gaagaacggc aactaagctg ccgggtttga aacacggatg atctcgcgga
    13021 gggtagcatg ttgattgtaa cgatgacaga gcgttgctgc ctgtgatcaa atatcatctc
    13081 cctcgcagag atccgaatta tcagccttct tattcatttc tcgcttaacc gtgacaggct
    13141 gtcgatcttg agaactatgc cgacataata ggaaatcgct ggataaagcc gctgaggaag
    13201 ctgagtggcg ctatttcttt agaagtgaac gttgacgata tcaactcccc tatccattgc
    13261 tcaccgaatg gtacaggtcg gggacccgaa gttccgactg tcggcctgat gcatccccgg
    13321 ctgatcgacc ccagatctgg ggctgagaaa gcccagtaag gaaacaactg taggttcgag
    13381 tcgcgagatc ccccggaacc aaaggaagta ggttaaaccc gctccgatca ggccgagcca
    13441 cgccaggccg agaacattgg ttcctgtagg catcgggatt ggcggatcaa acactaaagc
    13501 tactggaacg agcagaagtc ctccggccgc cagttgccag gcggtaaagg tgagcagagg
    13561 cacgggaggt tgccacttgc gggtcagcac ggttccgaac gccatggaaa ccgcccccgc
    13621 caggcccgct gcgacgccga caggatctag cgctgcgttt ggtgtcaaca ccaacagcgc
    13681 cacgcccgca gttccgcaaa tagcccccag gaccgccatc aatcgtatcg ggctacctag
    13741 cagagcggca gagatgaaca cgaccatcag cggctgcaca gcgcctaccg tcgccgcgac
    13801 cccgcccggc aggcggtaga ccgaaataaa caacaagctc cagaatagcg aaatattaag
    13861 tgcgccgagg atgaagatgc gcatccacca gattcccgtt ggaatctgtc ggacgatcat
    13921 cacgagcaat aaacccgccg gcaacgcccg cagcagcata ccggcgaccc ctcggcctcg
    13981 ctgttcgggc tccacgaaaa cgccggacag atgcgccttg tgagcgtcct tggggccgtc
    14041 ctcctgtttg aagaccgaca gcccaatgat ctcgccgtcg atgtaggcgc cgaatgccac
    14101 ggcatctcgc aaccgttcag cgaacgcctc catgggcttt ttctcctcgt gctcgtaaac
    14161 ggacccgaac atctctggag ctttcttcag ggccgacaat cggatctcgc ggaaatcctg
    14221 cacgtcggcc gctccaagcc gtcgaatctg agccttaatc acaattgtca attttaatcc
    14281 tctgtttatc ggcagttcgt agagcgcgcc gtgcgtcccg agcgatactg agcgaagcaa
    14341 gtgcgtcgag cagtgcccgc ttgttcctga aatgccagta aagcgctggc tgctgaaccc
    14401 ccagccggaa ctgaccccac aaggccctag cgtttgcaat gcaccaggtc atcattgacc
    14461 caggcgtgtt ccaccaggcc gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg
    14521 cgccacttct tcacgcgggt ggaatccgat ccgcacatga ggcggaaggt ttccagcttg
    14581 agcgggtacg gctcccggtg cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac
    14641 agcttgcggt acttctccca tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg
    14701 atttcctcgt cgatcaggac ctggcaacgg gacgttttct tgccacggtc caggacgcgg
    14761 aagcggtgca gcagcgacac cgattccagg tgcccaacgc ggtcggacgt gaagcccatc
    14821 gccgtcgcct gtaggcgcga caggcattcc tcggccttcg tgtaataccg gccattgatc
    14881 gaccagccca ggtcctggca aagctcgtag aacgtgaagg tgatcggctc gccgataggg
    14941 gtgcgcttcg cgtactccaa cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc
    15001 tcgacgccgg tgtaggtgat cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc
    15061 agcgcctcgc gcgggatttt cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg
    15121 tttggcatcg ctcgcatcgt gtccggccac ggcgcaatat cgaacaagga aagctgcatt
    15181 tccttgatct gctgcttcgt gtgtttcagc aacgcggcct gcttggcctc gctgacctgt
    15241 tttgccaggt cctcgccggc ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg
    15301 atggtcatcg acttcgccaa acctgccgcc tcctgttcga gacgacgcga acgctccacg
    15361 gcggccgatg gcgcgggcag ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg
    15421 gccgtagctt gctggaccat cgagccgacg gactggaagg tttcgcgggg cgcacgcatg
    15481 acggtgcggc ttgcgatggt ttcggcatcc tcggcggaaa accccgcgtc gatcagttct
    15541 tgcctgtatg ccttccggtc aaacgtccga ttcattcacc ctccttgcgg gattgccccg
    15601 actcacgccg gggcaatgtg cccttattcc tgatttgacc cgcctggtgc cttggtgtcc
    15661 agataatcca ccttatcggc aatgaagtcg gtcccgtaga ccgtctggcc gtccttctcg
    15721 tacttggtat tccgaatctt gccctgcacg aataccagcg accccttgcc caaatacttg
    15781 ccgtgggcct cggcctgaga gccaaaacac ttgatgcgga agaagtcggt gcgctcctgc
    15841 ttgtcgccgg catcgttgcg ccacatctag gtactaaaac aattcatcca gtaaaatata
    15901 atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata
    15961 ctgttcttcc ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt
    16021 gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa
    16081 gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt
    16141 ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc
    16201 gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc
    16261 taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc
    16321 cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag
    16381 gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag
    16441 gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac
    16501 atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc
    16561 tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta
    16621 tttttcgatc agttttttca attccggtga tattctcatt ttagccattt attatttcct
    16681 tcctcttttc tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc
    16741 aattcactgt tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg
    16801 ttttcaaagt tggcgtataa catagtatcg acggagccga ttttgaaacc acaattatgg
    16861 gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct
    16921 tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa
    16981 agcaccgccg gacatcagcg ctatctctgc tctcactgcc gtaaaacatg gcaactgcag
    17041 ttcacttaca ccgcttctca acccggtacg caccagaaaa tcattgatat ggccatgaat
    17101 ggcgttggat gccgggcaac agcccgcatt atgggcgttg gcctcaacac gattttacgt
    17161 cacttaaaaa actcaggccg cagtcggtaa cctcgcgcat acagccgggc agtgacgtca
    17221 tcgtctgcgc ggaaatggac gaacagtggg gctatgtcgg ggctaaatcg cgccagcgct
    17281 ggctgtttta cgcgtatgac agtctccgga agacggttgt tgcgcacgta ttcggtgaac
    17341 gcactatggc gacgctgggg cgtcttatga gcctgctgtc accctttgac gtggtgatat
    17401 ggatgacgga tggctggccg ctgtatgaat cccgcctgaa gggaaagctg cacgtaatca
    17461 gcaagcgata tacgcagcga attgagcggc ataacctgaa tctgaggcag cacctggcac
    17521 ggctgggacg gaagtcgctg tcgttctcaa aatcggtgga gctgcatgac aaagtcatcg
    17581 ggcattatct gaacataaaa cactatcaat aagttggagt cattacccaa ttatgataga
    17641 atttacaagc tataaggtta ttgtcctggg tttcaagcat tagtccatgc aagtttttat
    17701 gctttgccca ttctatagat atattgataa gcgcgctgcc tatgccttgc cccctgaaat
    17761 ccttacatac ggcgatatct tctatataaa agatatatta tcttatcagt attgtcaata
    17821 tattcaaggc aatctgcctc ctcatcctct tcatcctctt cgtcttggta gctttttaaa
    17881 tatggcgctt catagagtaa ttctgtaaag gtccaattct cgttttcata cctcggtata
    17941 atcttaccta tcacctcaaa tggttcgctg ggtttatcgc acccccgaac acgagcacgg
    18001 cacccgcgac cactatgcca agaatgccca aggtaaaaat tgccggcccc gccatgaagt
    18061 ccgtgaatgc cccgacggcc gaagtgaagg gcaggccgcc acccaggccg ccgccctcac
    18121 tgcccggcac ctggtcgctg aatgtcgatg ccagcacctg cggcacgtca atgcttccgg
    18181 gcgtcgcgct cgggctgatc gcccatcccg ttactgcccc gatcccggca atggcaagga
    18241 ctgccagcgc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg
    18301 gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc
    18361 ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt
    18421 ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg
    18481 caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat
    18541 ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg
    18601 cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg
    18661 aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc
    18721 ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa
    18781 gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa
    18841 cgccggcggc cggccgcggt gtctcgcaca cggcttcgac ggcgtttctg gcgcgtttgc
    18901 agggccatag acggccgcca gcccagcggc gagggcaacc agcccggtga gcgtcggaaa
    18961 ggcgctcggt cttgccttgc tcgtcggtg
//
