LOCUS       sequence-202614-       18424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g044580 (ERDJ2). Also 
            expresses Cas9 from rolD promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18424)
  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 18424)
  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: pSC15
FEATURES             Location/Qualifiers
     source          1..18424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(145..516)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            549..658
                     /note="incP origin of transfer"
     primer_bind     881..900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1134..1151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1339..1989)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2454..2478
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     2730..2746
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3267..3291
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3313..3360
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3367..3387
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3412..4848,5038..7701)
                     /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          4849..5037
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             7702..7749
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      7760..8012
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8057..8077
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8432..8507
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8519..8543)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9021..9096
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9108..9132)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9151..9175)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9213..9920
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     10711..10731
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             10808..11359
                     /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      11385..11637
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(11650..11667)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11650..11666)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11659..11681)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(11806..11825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12016..12037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12284..12308
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(13904..15052)
                     /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(15351..16145)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16337..17104
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      join(17797..18424,1..8)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tgagcgtcgg aaaggcgctc ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc
       61 gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc
      121 cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca cgcccatcat
      181 gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga ggatcgtggc
      241 atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca ggccgcccag
      301 gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc
      361 cgtgattttg tagccctggc cgacggccag caggtaggcc gacaggctca tgccggccgc
      421 cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct tgataggtgg
      481 gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc
      541 ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata
      601 agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg
      661 ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata
      721 tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta
      781 tgcagcggaa aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1021 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1081 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1141 cagtgagcga ggaagcggaa gagcgccaga aggccgccag agaggccgag cgcggccgtg
     1201 aggcttggac gctagggcag ggcatgaaaa agcccgtagc gggctgctac gggcgtctga
     1261 cgcggtggaa agggggaggg gatgttgtct acatggctct gctgtagtga gtgggttgcg
     1321 ctccggcagc ggtcctgatc aatcgtcacc ctttctcggt ccttcaacgt tcctgacaac
     1381 gagcctcctt ttcgccaatc catcgacaat caccgcgagt ccctgctcga acgctgcgtc
     1441 cggaccggct tcgtcgaagg cgtctatcgc ggcccgcaac agcggcgaga gcggagcctg
     1501 ttcaacggtg ccgccgcgct cgccggcatc gctgtcgccg gcctgctcct caagcacggc
     1561 cccaacagtg aagtagctga ttgtcatcag cgcattgacg gcgtccccgg ccgaaaaacc
     1621 cgcctcgcag aggaagcgaa gctgcgcgtc ggccgtttcc atctgcggtg cgcccggtcg
     1681 cgtgccggca tggatgcgcg cgccatcgcg gtaggcgagc agcgcctgcc tgaagctgcg
     1741 ggcattcccg atcagaaatg agcgccagtc gtcgtcggct ctcggcaccg aatgcgtatg
     1801 attctccgcc agcatggctt cggccagtgc gtcgagcagc gcccgcttgt tcctgaagtg
     1861 ccagtaaagc gccggctgct gaacccccaa ccgttccgcc agtttgcgtg tcgtcagacc
     1921 gtctacgccg acctcgttca acaggtccag ggcggcacgg atcactgtat tcggctgcaa
     1981 ctttgtcatg cttgacactt tatcactgat aaacataata tgtccaccaa cttatcagtg
     2041 ataaagaatc cgcgcgttca atcggaccag cggaggctgg tccggaggcc agacgtgaaa
     2101 cccaacatac ccctgatcgt aattctgagc actgtcgcgc tcgacgctgt cggcatcggc
     2161 ctgattatgc cggtgctgcc gggcctcctg cgcgatctgg ttcactcgaa cgacgtcacc
     2221 gcccactatg gcattctgct ggcgctgtat gcgttggtgc aatttgcctg cgcacctgtg
     2281 ctgggcgcgc tgtcggatcg tttcgggcgg cggccaatct tgctcgtctc gctggccggc
     2341 gccagatctg gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
     2401 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
     2461 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
     2521 tcatgagcgg agaattaagg gagtcacgtt atgacccccg ccgatgacgc gggacaagcc
     2581 gttttacgtt tggaactgac agaaccgcaa cgttgaagga gccactcagc cgcgggtttc
     2641 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
     2701 taaggtcact atcagctagc gcggccgcgc gaggtcgacg gtatcgataa gcttgatatc
     2761 gaattcctgc agcccggggg atccgcccgc tctagactgc cttgtaggtg cgaattttct
     2821 ctgctcaaat tgttgaggtt agcggatttg taaacgcgtt tatatgggct gcttggaggg
     2881 tacttttgga ttaatttttt tctgccagcg cattctgacg cggcaccgct ttggaaagtg
     2941 cgctgtgggt ccgcgttttc tacaataatg tgccgatccg gtcagaaagt atatggatga
     3001 gttgtgccag cctcaccaac gtgctgcagg cccatcatga ctacttcaat gttaatgggg
     3061 gtaatgaata aataggcgaa attgggttca cggtgggccc agggaatata atattgccgc
     3121 agaggtagtc ggatgccaag gcccgcaact aatagttcac gaacaaattc attgtagtgg
     3181 gcggccaact ccaaaaccaa ttgccagtta ttgtattgca atacatatat ggcttcctcc
     3241 ttgctcgagg cgcgccaagc tatcaaacaa gtttgtacaa aaaagcaggc tccgaattcg
     3301 cccttcacca tggattacaa ggatgatgat gataaggatt acaaggatga tgatgataag
     3361 atggctccaa agaagaagag aaaggttgga atccacggag ttccagctgc tgataagaag
     3421 tactctatcg gacttgacat cggaaccaac tctgttggat gggctgttat caccgatgag
     3481 tacaaggttc catctaagaa gttcaaggtt cttggaaaca ccgatagaca ctctatcaag
     3541 aagaacctta tcggtgctct tcttttcgat tctggagaga ccgctgaggc taccagattg
     3601 aagagaaccg ctagaagaag atacaccaga agaaagaaca gaatctgcta ccttcaggaa
     3661 atcttctcta acgagatggc taaggttgat gattctttct tccacagact tgaggagtct
     3721 ttccttgttg aggaggataa gaagcacgag agacacccaa tcttcggaaa catcgttgat
     3781 gaggttgctt accacgagaa gtacccaacc atctaccacc ttagaaagaa gttggttgat
     3841 tctaccgata aggctgatct tagacttatc taccttgctc ttgctcacat gatcaagttc
     3901 agaggacact tccttatcga gggagacctt aacccagata actctgatgt tgataagttg
     3961 ttcatccagc ttgttcagac ctacaaccag cttttcgagg agaacccaat caacgcttct
     4021 ggagttgatg ctaaggctat cctttctgct agactttcta agtctcgtag acttgagaac
     4081 cttatcgctc agcttccagg agagaagaag aacggacttt tcggaaacct tatcgctctt
     4141 tctcttggac ttaccccaaa cttcaagtct aacttcgatc ttgctgagga tgctaagttg
     4201 cagctttcta aggataccta cgatgatgat cttgataacc ttcttgctca gatcggagat
     4261 cagtacgctg atcttttcct tgctgctaag aacctttctg atgctatcct tctttctgac
     4321 atccttagag ttaacaccga gatcaccaag gctccacttt ctgcttctat gatcaagaga
     4381 tacgatgagc accaccagga tcttaccctt ttgaaggctc ttgttagaca gcagcttcca
     4441 gagaagtaca aggaaatctt cttcgatcag tctaagaacg gatacgctgg atacatcgat
     4501 ggaggagctt ctcaggagga gttctacaag ttcatcaagc caatccttga gaagatggat
     4561 ggaaccgagg agcttcttgt taagttgaac agagaggatc ttcttagaaa gcagagaacc
     4621 ttcgataacg gatctatccc acaccagatc caccttggag agcttcacgc tatccttcgt
     4681 agacaggagg atttctaccc attcttgaag gataacagag agaagatcga gaagatcctt
     4741 accttcagaa tcccatacta cgttggacca cttgctagag gaaactctcg tttcgcttgg
     4801 atgaccagaa agtctgagga gaccatcacc ccttggaact tcgaggaggt aagtttctgc
     4861 ttctaccttt gatatatata taataattat cattaattag tagtaatata atatttcaaa
     4921 tatttttttc aaaataaaag aatgtagtat atagcaattg cttttctgta gtttataagt
     4981 gtgtatattt taatttataa cttttctaat atatgaccaa aatttgttga tgtgcaggtt
     5041 gttgataagg gagcttctgc tcagtctttc atcgagagaa tgaccaactt cgataagaac
     5101 cttccaaacg agaaggttct tccaaagcac tctcttcttt acgagtactt caccgtttac
     5161 aacgagctta ccaaggttaa gtacgttacc gagggaatga gaaagccagc tttcctttct
     5221 ggagagcaga agaaggctat cgttgatctt cttttcaaga ccaacagaaa ggttaccgtt
     5281 aagcagttga aggaggatta cttcaagaag atcgagtgct tcgattctgt tgaaatctct
     5341 ggagttgagg atagattcaa cgcttctctt ggaacctacc acgatctttt gaagatcatc
     5401 aaggataagg atttccttga taacgaggag aacgaggaca tccttgagga catcgttctt
     5461 acccttaccc ttttcgagga tagagagatg atcgaggaga gactcaagac ctacgctcac
     5521 cttttcgatg ataaggttat gaagcagttg aagagaagaa gatacaccgg atggggtaga
     5581 ctttctcgta agttgatcaa cggaatcaga gataagcagt ctggaaagac catccttgat
     5641 ttcttgaagt ctgatggatt cgctaacaga aacttcatgc agcttatcca cgatgattct
     5701 cttaccttca aggaggacat ccagaaggct caggtttctg gacagggaga ttctcttcac
     5761 gagcacatcg ctaaccttgc tggatctcca gctatcaaga agggaatcct tcagaccgtt
     5821 aaggttgttg atgagcttgt taaggttatg ggtagacaca agccagagaa catcgttatc
     5881 gagatggcta gagagaacca gaccacccag aagggacaga agaactctcg tgagagaatg
     5941 aagagaatcg aggagggaat caaggagctt ggatctcaaa tcttgaagga gcacccagtt
     6001 gagaacaccc agcttcagaa cgagaagttg tacctttact accttcagaa cggaagagat
     6061 atgtacgttg atcaggagct tgacatcaac agactttctg attacgatgt tgatcacatc
     6121 gttccacagt ctttcttgaa ggatgattct atcgataaca aggttcttac ccgttctgat
     6181 aagaacagag gaaagtctga taacgttcca tctgaggagg ttgttaagaa gatgaagaac
     6241 tactggagac agcttcttaa cgctaagttg atcacccaga gaaagttcga taaccttacc
     6301 aaggctgaga gaggaggact ttctgagctt gataaggctg gattcatcaa gagacagctt
     6361 gttgagacca gacagatcac caagcacgtt gctcagatcc ttgattctcg tatgaacacc
     6421 aagtacgatg agaacgataa gttgatcaga gaggttaagg ttatcacctt gaagtctaag
     6481 ttggtttctg atttcagaaa ggatttccag ttctacaagg ttagagagat caacaactac
     6541 caccacgctc acgatgctta ccttaacgct gttgttggaa ccgctcttat caagaagtac
     6601 ccaaagttgg agtctgagtt cgtttacgga gattacaagg tttacgatgt tagaaagatg
     6661 atcgctaagt ctgagcagga gatcggaaag gctaccgcta agtacttctt ctactctaac
     6721 atcatgaact tcttcaagac cgagatcacc cttgctaacg gagagatcag aaagagacca
     6781 cttatcgaga ccaacggaga gaccggagag atcgtttggg ataagggaag agatttcgct
     6841 accgttagaa aggttctttc tatgccacag gttaacatcg ttaagaaaac cgaggttcag
     6901 accggaggat tctctaagga gtctatcctt ccaaagagaa actctgataa gttgatcgct
     6961 agaaagaagg attgggaccc aaagaagtac ggaggattcg attctccaac cgttgcttac
     7021 tctgttcttg ttgttgctaa ggttgagaag ggaaagtcta agaagttgaa gtctgttaag
     7081 gagcttcttg gaatcaccat catggagcgt tcttctttcg agaagaaccc aatcgatttc
     7141 cttgaggcta agggatacaa ggaggttaag aaggatctta tcatcaagtt gccaaagtac
     7201 tctcttttcg agcttgagaa cggaagaaag agaatgcttg cttctgctgg agagcttcag
     7261 aagggaaacg agcttgctct tccatctaag tacgttaact tcctttacct tgcttctcac
     7321 tacgagaagt tgaagggatc tccagaggat aacgagcaga agcagctttt cgttgagcag
     7381 cacaagcact accttgatga gatcatcgag caaatctctg agttctctaa gagagttatc
     7441 cttgctgatg ctaaccttga taaggttctt tctgcttaca acaagcacag agataagcca
     7501 atcagagagc aggctgagaa catcatccac cttttcaccc ttaccaacct tggtgctcca
     7561 gctgctttca agtacttcga taccaccatc gatagaaaaa gatacacctc taccaaggag
     7621 gttcttgatg ctacccttat ccaccagtct atcaccggac tttacgagac cagaatcgat
     7681 ctttctcagc ttggaggaga taagagacca gctgctacca agaaggctgg acaggctaag
     7741 aagaagaagt gagacgtccg atcgttcaaa catttggcaa taaagtttct taagattgaa
     7801 tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg ttaagcatgt
     7861 aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga ttagagtccc
     7921 gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact aggataaatt
     7981 atcgcgcgcg gtgtcatcta tgttactaga tcgggaattg atcccccctc gacagcttcc
     8041 ggaaagggcg aattcgcaac tttgtataca aaagttgccg agctcgcggc cgcatgttgt
     8101 tgttaccaga aagtaaataa atgttcaatc tctgatgttc tcaagtaagt gagttttatt
     8161 gggaataata ttaacttatg ttcttcttgc atttgatttc tttgccgctc tcttcttcta
     8221 tcttaaatct gtgtatacta tttcactatt gggcttttta ttagtctata atgggactca
     8281 aaataaggct ttggcccaca tcaaaaagat aagtcacaaa tcaaaactaa attcagagtc
     8341 ttttctccca catcggtcac tgtactcatt ttgtgtttgt ttatatatta cacgaaccga
     8401 tctttggtac ggtactgaag gttagcgacg agttttagag ctagaaatag caagttaaaa
     8461 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttctagac
     8521 ccagctttct tgtacaaagt tggcattacc atggcttcgt tgaacaacgg aaactcgact
     8581 tgccttccgc acaatacatc atttcttctt agcttttttt cttcttcttc gttcatacag
     8641 tttttttttg tttatcagct tacattttct tgaaccgtag ctttcgtttt cttcttttta
     8701 actttccatt cggagttttt gtatcttgtt tcatagtttg tcccaggatt agaatgatta
     8761 ggcatcgaac cttcaagaat ttgattgaat aaaacatctt cattcttaag atatgaagat
     8821 aatcttcaaa aggcccctgg gaatctgaaa gaagagaagc aggcccattt atatgggaaa
     8881 gaacaatagt atttcttata taggcccatt taagttgaaa acaatcttca aaagtcccac
     8941 atcgcttaga taagaaaacg aagctgagtt tatatacagc tagagtcgaa gtagtgattg
     9001 aatattgatg gggcatctgg gttttagagc tagaaatagc aagttaaaat aaggctagtc
     9061 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctagacc cagctttctt
     9121 gtacaaagtt ggcattactg caggtcgacg acccagcttt cttgtacaaa gtggttcgat
     9181 aattcttaat taactagttc taatctagag tcctgcttta atgagatatg cgagacgcct
     9241 atgatcgcat gatatttgct ttcaattctg ttgtgcacgt tgtaaaaaac ctgagcatgt
     9301 gtagctcaga tccttaccgc cggtttcggt tcattctaat gaatatatca cccgttacta
     9361 tcgtattttt atgaataata ttctccgttc aatttactga ttgtacccta ctacttatat
     9421 gtacaatatt aaaatgaaaa caatatattg tgctgaatag gtttatagcg acatctatga
     9481 tagagcgcca caataacaaa caattgcgtt ttattattac aaatccaatt ttaaaaaaag
     9541 cggcagaacc ggtcaaacct aaaagactga ttacataaat cttattcaaa tttcaaaagt
     9601 gccccagggg ctagtatcta cgacacaccg agcggcgaac taataacgct cactgaaggg
     9661 aactccggtt ccccgccggc gcgcatgggt gagattcctt gaagttgagt attggccgtc
     9721 cgctctaccg aaagttacgg gcaccattca acccggtcca gcacggcggc cgggtaaccg
     9781 acttgctgcc ccgagaatta tgcagcattt ttttggtgta tgtgggcccc aaatgaagtg
     9841 caggtcaaac cttgacagtg acgacaaatc gttgggcggg tccagggcga attttgcgac
     9901 aacatgtcga ggctcagcag gacctgcagg tccccagatt agccttttca atttcagaaa
     9961 gaatgctaac ccacagatgg ttagagaggc ttacgcagca ggtctcatca agacgatcta
    10021 cccgagcaat aatctccagg aaatcaaata ccttcccaag aaggttaaag atgcagtcaa
    10081 aagattcagg actaactgca tcaagaacac agagaaagat atatttctca agatcagaag
    10141 tactattcca gtatggacga ttcaaggctt gcttcacaaa ccaaggcaag taatagagat
    10201 tggagtctct aaaaaggtag ttcccactga atcaaaggcc atggagtcaa agattcaaat
    10261 agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga gtctcttacg
    10321 actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac ttgtctactc
    10381 caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt ttcaacaaag
    10441 ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact ttattgtgaa
    10501 gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggccat
    10561 cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
    10621 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
    10681 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
    10741 aggaagttca tttcatataa ggaagttcat ttcatttgga gagaacacgg gggactctag
    10801 aggatccatg agcccagaac gacgcccggc cgacatccgc cgtgccaccg aggcggacat
    10861 gccggcggtc tgcaccatcg tcaaccacta catcgagaca agcacggtca acttccgtac
    10921 cgagccgcag gaaccgcagg agtggacgga cgacctcgtc cgtctgcggg agcgctatcc
    10981 ctggctcgtc gccgaggtgg acggcgaggt cgccggcatc gcctacgcgg gcccctggaa
    11041 ggcacgcaac gcctacgact ggacggccga gtcgaccgtg tacgtctccc cccgccacca
    11101 gcggacggga ctgggctcca cgctctacac ccacctgctg aagtccctgg aggcacaggg
    11161 cttcaagagc gtggtcgctg tcatcgggct gcccaacgac ccgagcgtgc gcatgcacga
    11221 ggcgctcgga tatgcccccc gcggcatgct gcgggcggcc ggcttcaagc acgggaactg
    11281 gcatgacgtg ggtttctggc agctggactt cagcctgccg gtaccgcccc gtccggtcct
    11341 gcccgtcacc gagatctgaa ggggtaccga gctcgaattt ccccgatcgt tcaaacattt
    11401 ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat
    11461 ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga
    11521 gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa
    11581 tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg
    11641 aattaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
    11701 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
    11761 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gcccgctcct ttcgctttct
    11821 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
    11881 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
    11941 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
    12001 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
    12061 gctattcttt tgatttataa gggattttgc cgatttcgga accaccatca aacaggattt
    12121 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
    12181 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc cagtacatta
    12241 aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata
    12301 tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg
    12361 atacaggcag cccatcagtc cgggacggcg tcagcgggag agccgttgta aggcggcaga
    12421 ctttgctcat gttaccgatg ctattcggaa gaacggcaac taagctgccg ggtttgaaac
    12481 acggatgatc tcgcggaggg tagcatgttg attgtaacga tgacagagcg ttgctgcctg
    12541 tgatcaaata tcatctccct cgcagagatc cgaattatca gccttcttat tcatttctcg
    12601 cttaaccgtg acaggctgtc gatcttgaga actatgccga cataatagga aatcgctgga
    12661 taaagccgct gaggaagctg agtggcgcta tttctttaga agtgaacgtt gacgatatca
    12721 actcccctat ccattgctca ccgaatggta caggtcgggg acccgaagtt ccgactgtcg
    12781 gcctgatgca tccccggctg atcgacccca gatctggggc tgagaaagcc cagtaaggaa
    12841 acaactgtag gttcgagtcg cgagatcccc cggaaccaaa ggaagtaggt taaacccgct
    12901 ccgatcaggc cgagccacgc caggccgaga acattggttc ctgtaggcat cgggattggc
    12961 ggatcaaaca ctaaagctac tggaacgagc agaagtcctc cggccgccag ttgccaggcg
    13021 gtaaaggtga gcagaggcac gggaggttgc cacttgcggg tcagcacggt tccgaacgcc
    13081 atggaaaccg cccccgccag gcccgctgcg acgccgacag gatctagcgc tgcgtttggt
    13141 gtcaacacca acagcgccac gcccgcagtt ccgcaaatag cccccaggac cgccatcaat
    13201 cgtatcgggc tacctagcag agcggcagag atgaacacga ccatcagcgg ctgcacagcg
    13261 cctaccgtcg ccgcgacccc gcccggcagg cggtagaccg aaataaacaa caagctccag
    13321 aatagcgaaa tattaagtgc gccgaggatg aagatgcgca tccaccagat tcccgttgga
    13381 atctgtcgga cgatcatcac gagcaataaa cccgccggca acgcccgcag cagcataccg
    13441 gcgacccctc ggcctcgctg ttcgggctcc acgaaaacgc cggacagatg cgccttgtga
    13501 gcgtccttgg ggccgtcctc ctgtttgaag accgacagcc caatgatctc gccgtcgatg
    13561 taggcgccga atgccacggc atctcgcaac cgttcagcga acgcctccat gggctttttc
    13621 tcctcgtgct cgtaaacgga cccgaacatc tctggagctt tcttcagggc cgacaatcgg
    13681 atctcgcgga aatcctgcac gtcggccgct ccaagccgtc gaatctgagc cttaatcaca
    13741 attgtcaatt ttaatcctct gtttatcggc agttcgtaga gcgcgccgtg cgtcccgagc
    13801 gatactgagc gaagcaagtg cgtcgagcag tgcccgcttg ttcctgaaat gccagtaaag
    13861 cgctggctgc tgaaccccca gccggaactg accccacaag gccctagcgt ttgcaatgca
    13921 ccaggtcatc attgacccag gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg
    13981 cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga atccgatccg cacatgaggc
    14041 ggaaggtttc cagcttgagc gggtacggct cccggtgcga gctgaaatag tcgaacatcc
    14101 gtcgggccgt cggcgacagc ttgcggtact tctcccatat gaatttcgtg tagtggtcgc
    14161 cagcaaacag cacgacgatt tcctcgtcga tcaggacctg gcaacgggac gttttcttgc
    14221 cacggtccag gacgcggaag cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt
    14281 cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt
    14341 aataccggcc attgatcgac cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga
    14401 tcggctcgcc gataggggtg cgcttcgcgt actccaacac ctgctgccac accagttcgt
    14461 catcgtcggc ccgcagctcg acgccggtgt aggtgatctt cacgtccttg ttgacgtgga
    14521 aaatgacctt gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg
    14581 cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga
    14641 acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct
    14701 tggcctcgct gacctgtttt gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca
    14761 tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc tgttcgagac
    14821 gacgcgaacg ctccacggcg gccgatggcg cgggcagggc agggggagcc agttgcacgc
    14881 tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga gccgacggac tggaaggttt
    14941 cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc
    15001 ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc attcaccctc
    15061 cttgcgggat tgccccgact cacgccgggg caatgtgccc ttattcctga tttgacccgc
    15121 ctggtgcctt ggtgtccaga taatccacct tatcggcaat gaagtcggtc ccgtagaccg
    15181 tctggccgtc cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat accagcgacc
    15241 ccttgcccaa atacttgccg tgggcctcgg cctgagagcc aaaacacttg atgcggaaga
    15301 agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca catctaggta ctaaaacaat
    15361 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
    15421 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag
    15481 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
    15541 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
    15601 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
    15661 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
    15721 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
    15781 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
    15841 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
    15901 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
    15961 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
    16021 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
    16081 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
    16141 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
    16201 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
    16261 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
    16321 tgaaaccaca attatgggtg atgctgccaa cttactgatt tagtgtatga tggtgttttt
    16381 gaggtgctcc agtggcttct gtttctatca gctgtccctc ctgttcagct actgacgggg
    16441 tggtgcgtaa cggcaaaagc accgccggac atcagcgcta tctctgctct cactgccgta
    16501 aaacatggca actgcagttc acttacaccg cttctcaacc cggtacgcac cagaaaatca
    16561 ttgatatggc catgaatggc gttggatgcc gggcaacagc ccgcattatg ggcgttggcc
    16621 tcaacacgat tttacgtcac ttaaaaaact caggccgcag tcggtaacct cgcgcataca
    16681 gccgggcagt gacgtcatcg tctgcgcgga aatggacgaa cagtggggct atgtcggggc
    16741 taaatcgcgc cagcgctggc tgttttacgc gtatgacagt ctccggaaga cggttgttgc
    16801 gcacgtattc ggtgaacgca ctatggcgac gctggggcgt cttatgagcc tgctgtcacc
    16861 ctttgacgtg gtgatatgga tgacggatgg ctggccgctg tatgaatccc gcctgaaggg
    16921 aaagctgcac gtaatcagca agcgatatac gcagcgaatt gagcggcata acctgaatct
    16981 gaggcagcac ctggcacggc tgggacggaa gtcgctgtcg ttctcaaaat cggtggagct
    17041 gcatgacaaa gtcatcgggc attatctgaa cataaaacac tatcaataag ttggagtcat
    17101 tacccaatta tgatagaatt tacaagctat aaggttattg tcctgggttt caagcattag
    17161 tccatgcaag tttttatgct ttgcccattc tatagatata ttgataagcg cgctgcctat
    17221 gccttgcccc ctgaaatcct tacatacggc gatatcttct atataaaaga tatattatct
    17281 tatcagtatt gtcaatatat tcaaggcaat ctgcctcctc atcctcttca tcctcttcgt
    17341 cttggtagct ttttaaatat ggcgcttcat agagtaattc tgtaaaggtc caattctcgt
    17401 tttcatacct cggtataatc ttacctatca cctcaaatgg ttcgctgggt ttatcgcacc
    17461 cccgaacacg agcacggcac ccgcgaccac tatgccaaga atgcccaagg taaaaattgc
    17521 cggccccgcc atgaagtccg tgaatgcccc gacggccgaa gtgaagggca ggccgccacc
    17581 caggccgccg ccctcactgc ccggcacctg gtcgctgaat gtcgatgcca gcacctgcgg
    17641 cacgtcaatg cttccgggcg tcgcgctcgg gctgatcgcc catcccgtta ctgccccgat
    17701 cccggcaatg gcaaggactg ccagcgctgc catttttggg gtgaggccgt tcgcggccga
    17761 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag
    17821 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa
    17881 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa
    17941 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca
    18001 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc
    18061 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg
    18121 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg
    18181 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg
    18241 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt
    18301 ccgcgaggta tccacaacgc cggcggccgg ccgcggtgtc tcgcacacgg cttcgacggc
    18361 gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc
    18421 ccgg
//
