LOCUS       sequence-202766-       18989 bp    DNA     circular SYN 15-APR-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101180 (Cop-like). Also 
            expresses Cas9 from Gmubi promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18989)
  AUTHORS   Curtin SJ, Tiffin P, Guhlin J, Trujillo DI, Burghart LT, Atkins P, 
            Baltes NJ, Denny R, Voytas DF, Stupar RM, Young ND
  TITLE     Validating Genome-Wide Association Candidates Controlling 
            Quantitative Variation in Nodulation.
  JOURNAL   Plant Physiol. 2017 Feb;173(2):921-931. doi: 10.1104/pp.16.01923. 
            Epub 2017 Jan 5.
  PUBMED    28057894
REFERENCE   2  (bases 1 to 18989)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pSC39
FEATURES             Location/Qualifiers
     source          1..18989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(104..475)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            508..617
                     /note="incP origin of transfer"
     primer_bind     840..859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1093..1110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1298..1948)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2413..2437
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3645..3669
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3691..3738
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3745..3765
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3790..5226,5416..8079)
                     /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          5227..5415
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8080..8127
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8138..8390
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8435..8455
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8810..8885
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8897..8921)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9399..9474
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9486..9510)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9529..9553)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9591..10298
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        10848..11193
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     11136..11156
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11214..11765
                     /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      11909..12161
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12174..12191)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12174..12190)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12183..12205)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12330..12349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12540..12561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12808..12832
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14428..15576)
                     /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(15875..16669)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16861..17628
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      18251..18966
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gatgtacttc accagctccg cgaagtcgct cttcttgatg gagcgcatgg ggacgtgctt
       61 ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc tctgccctcg
      121 ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg atcttcgcca
      181 gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg gtgagccaga
      241 gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc tcgcggacgt
      301 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg
      361 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc
      421 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct
      481 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga
      541 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta
      601 cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct acacgaaccc
      661 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa
      721 tgaccccgaa gcagggttat gcagcggaaa agcgccacgc ttcccgaagg gagaaaggcg
      781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
      901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
      961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgccagaa ggccgccaga
     1141 gaggccgagc gcggccgtga ggcttggacg ctagggcagg gcatgaaaaa gcccgtagcg
     1201 ggctgctacg ggcgtctgac gcggtggaaa gggggagggg atgttgtcta catggctctg
     1261 ctgtagtgag tgggttgcgc tccggcagcg gtcctgatca atcgtcaccc tttctcggtc
     1321 cttcaacgtt cctgacaacg agcctccttt tcgccaatcc atcgacaatc accgcgagtc
     1381 cctgctcgaa cgctgcgtcc ggaccggctt cgtcgaaggc gtctatcgcg gcccgcaaca
     1441 gcggcgagag cggagcctgt tcaacggtgc cgccgcgctc gccggcatcg ctgtcgccgg
     1501 cctgctcctc aagcacggcc ccaacagtga agtagctgat tgtcatcagc gcattgacgg
     1561 cgtccccggc cgaaaaaccc gcctcgcaga ggaagcgaag ctgcgcgtcg gccgtttcca
     1621 tctgcggtgc gcccggtcgc gtgccggcat ggatgcgcgc gccatcgcgg taggcgagca
     1681 gcgcctgcct gaagctgcgg gcattcccga tcagaaatga gcgccagtcg tcgtcggctc
     1741 tcggcaccga atgcgtatga ttctccgcca gcatggcttc ggccagtgcg tcgagcagcg
     1801 cccgcttgtt cctgaagtgc cagtaaagcg ccggctgctg aacccccaac cgttccgcca
     1861 gtttgcgtgt cgtcagaccg tctacgccga cctcgttcaa caggtccagg gcggcacgga
     1921 tcactgtatt cggctgcaac tttgtcatgc ttgacacttt atcactgata aacataatat
     1981 gtccaccaac ttatcagtga taaagaatcc gcgcgttcaa tcggaccagc ggaggctggt
     2041 ccggaggcca gacgtgaaac ccaacatacc cctgatcgta attctgagca ctgtcgcgct
     2101 cgacgctgtc ggcatcggcc tgattatgcc ggtgctgccg ggcctcctgc gcgatctggt
     2161 tcactcgaac gacgtcaccg cccactatgg cattctgctg gcgctgtatg cgttggtgca
     2221 atttgcctgc gcacctgtgc tgggcgcgct gtcggatcgt ttcgggcggc ggccaatctt
     2281 gctcgtctcg ctggccggcg ccagatctgg ggaaccctgt ggttggcatg cacatacaaa
     2341 tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct aataaacgct
     2401 cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt taaactgaag
     2461 gcgggaaacg acaatctgat catgagcgga gaattaaggg agtcacgtta tgacccccgc
     2521 cgatgacgcg ggacaagccg ttttacgttt ggaactgaca gaaccgcaac gttgaaggag
     2581 ccactcagcc gcgggtttct ggagtttaat gagctaagca catacgtcag aaaccattat
     2641 tgcgcgttca aaagtcgcct aaggtcacta tcagctagac ggacgcggcc gcgggcccaa
     2701 tataacaacg acgtcgtaac agataaagcg aagcttgaag gtgcatgtga ctccgtcaag
     2761 attacgaaac cgccaactac cacgcaaatt gcaattctca atttcctaga aggactctcc
     2821 gaaaatgcat ccaataccaa atattacccg tgtcataggc accaagtgac accatacatg
     2881 aacacgcgtc acaatatgac tggagaaggg ttccacacct tatgctataa aacgccccac
     2941 acccctcctc cttccttcgc agttcaattc caatatattc cattctctct gtgtatttcc
     3001 ctacctctcc cttcaaggtt agtcgatttc ttctgttttt cttcttcgtt ctttccatga
     3061 attgtgtatg ttctttgatc aatacgatgt tgatttgatt gtgttttgtt tggtttcatc
     3121 gatcttcaat tttcataatc agattcagct tttattatct ttacaacaac gtccttaatt
     3181 tgatgattct ttaatcgtag atttgctcta attagagctt tttcatgtca gatcccttta
     3241 caacaagcct taattgttga ttcattaatc gtagattagg gcttttttca ttgattactt
     3301 cagatccgtt aaacgtaacc atagatcagg gctttttcat gaattacttc agatccgtta
     3361 aacaacagcc ttatttttta tacttctgtg gtttttcaag aaattgttca gatccgttga
     3421 caaaaagcct tattcgttga ttctatatcg tttttcgaga gatattgctc agatctgtta
     3481 gcaactgcct tgtttgttga ttctattgcc gtggattagg gttttttttc acgagattgc
     3541 ttcagatccg tacttaagat tacgtaatgg attttgattc tgatttatct gtgattgttg
     3601 actcgacagg taccttcaaa cctcgaggcg cgccaagcta tcaaacaagt ttgtacaaaa
     3661 aagcaggctc cgaattcgcc cttcaccatg gattacaagg atgatgatga taaggattac
     3721 aaggatgatg atgataagat ggctccaaag aagaagagaa aggttggaat ccacggagtt
     3781 ccagctgctg ataagaagta ctctatcgga cttgacatcg gaaccaactc tgttggatgg
     3841 gctgttatca ccgatgagta caaggttcca tctaagaagt tcaaggttct tggaaacacc
     3901 gatagacact ctatcaagaa gaaccttatc ggtgctcttc ttttcgattc tggagagacc
     3961 gctgaggcta ccagattgaa gagaaccgct agaagaagat acaccagaag aaagaacaga
     4021 atctgctacc ttcaggaaat cttctctaac gagatggcta aggttgatga ttctttcttc
     4081 cacagacttg aggagtcttt ccttgttgag gaggataaga agcacgagag acacccaatc
     4141 ttcggaaaca tcgttgatga ggttgcttac cacgagaagt acccaaccat ctaccacctt
     4201 agaaagaagt tggttgattc taccgataag gctgatctta gacttatcta ccttgctctt
     4261 gctcacatga tcaagttcag aggacacttc cttatcgagg gagaccttaa cccagataac
     4321 tctgatgttg ataagttgtt catccagctt gttcagacct acaaccagct tttcgaggag
     4381 aacccaatca acgcttctgg agttgatgct aaggctatcc tttctgctag actttctaag
     4441 tctcgtagac ttgagaacct tatcgctcag cttccaggag agaagaagaa cggacttttc
     4501 ggaaacctta tcgctctttc tcttggactt accccaaact tcaagtctaa cttcgatctt
     4561 gctgaggatg ctaagttgca gctttctaag gatacctacg atgatgatct tgataacctt
     4621 cttgctcaga tcggagatca gtacgctgat cttttccttg ctgctaagaa cctttctgat
     4681 gctatccttc tttctgacat ccttagagtt aacaccgaga tcaccaaggc tccactttct
     4741 gcttctatga tcaagagata cgatgagcac caccaggatc ttaccctttt gaaggctctt
     4801 gttagacagc agcttccaga gaagtacaag gaaatcttct tcgatcagtc taagaacgga
     4861 tacgctggat acatcgatgg aggagcttct caggaggagt tctacaagtt catcaagcca
     4921 atccttgaga agatggatgg aaccgaggag cttcttgtta agttgaacag agaggatctt
     4981 cttagaaagc agagaacctt cgataacgga tctatcccac accagatcca ccttggagag
     5041 cttcacgcta tccttcgtag acaggaggat ttctacccat tcttgaagga taacagagag
     5101 aagatcgaga agatccttac cttcagaatc ccatactacg ttggaccact tgctagagga
     5161 aactctcgtt tcgcttggat gaccagaaag tctgaggaga ccatcacccc ttggaacttc
     5221 gaggaggtaa gtttctgctt ctacctttga tatatatata ataattatca ttaattagta
     5281 gtaatataat atttcaaata tttttttcaa aataaaagaa tgtagtatat agcaattgct
     5341 tttctgtagt ttataagtgt gtatatttta atttataact tttctaatat atgaccaaaa
     5401 tttgttgatg tgcaggttgt tgataaggga gcttctgctc agtctttcat cgagagaatg
     5461 accaacttcg ataagaacct tccaaacgag aaggttcttc caaagcactc tcttctttac
     5521 gagtacttca ccgtttacaa cgagcttacc aaggttaagt acgttaccga gggaatgaga
     5581 aagccagctt tcctttctgg agagcagaag aaggctatcg ttgatcttct tttcaagacc
     5641 aacagaaagg ttaccgttaa gcagttgaag gaggattact tcaagaagat cgagtgcttc
     5701 gattctgttg aaatctctgg agttgaggat agattcaacg cttctcttgg aacctaccac
     5761 gatcttttga agatcatcaa ggataaggat ttccttgata acgaggagaa cgaggacatc
     5821 cttgaggaca tcgttcttac ccttaccctt ttcgaggata gagagatgat cgaggagaga
     5881 ctcaagacct acgctcacct tttcgatgat aaggttatga agcagttgaa gagaagaaga
     5941 tacaccggat ggggtagact ttctcgtaag ttgatcaacg gaatcagaga taagcagtct
     6001 ggaaagacca tccttgattt cttgaagtct gatggattcg ctaacagaaa cttcatgcag
     6061 cttatccacg atgattctct taccttcaag gaggacatcc agaaggctca ggtttctgga
     6121 cagggagatt ctcttcacga gcacatcgct aaccttgctg gatctccagc tatcaagaag
     6181 ggaatccttc agaccgttaa ggttgttgat gagcttgtta aggttatggg tagacacaag
     6241 ccagagaaca tcgttatcga gatggctaga gagaaccaga ccacccagaa gggacagaag
     6301 aactctcgtg agagaatgaa gagaatcgag gagggaatca aggagcttgg atctcaaatc
     6361 ttgaaggagc acccagttga gaacacccag cttcagaacg agaagttgta cctttactac
     6421 cttcagaacg gaagagatat gtacgttgat caggagcttg acatcaacag actttctgat
     6481 tacgatgttg atcacatcgt tccacagtct ttcttgaagg atgattctat cgataacaag
     6541 gttcttaccc gttctgataa gaacagagga aagtctgata acgttccatc tgaggaggtt
     6601 gttaagaaga tgaagaacta ctggagacag cttcttaacg ctaagttgat cacccagaga
     6661 aagttcgata accttaccaa ggctgagaga ggaggacttt ctgagcttga taaggctgga
     6721 ttcatcaaga gacagcttgt tgagaccaga cagatcacca agcacgttgc tcagatcctt
     6781 gattctcgta tgaacaccaa gtacgatgag aacgataagt tgatcagaga ggttaaggtt
     6841 atcaccttga agtctaagtt ggtttctgat ttcagaaagg atttccagtt ctacaaggtt
     6901 agagagatca acaactacca ccacgctcac gatgcttacc ttaacgctgt tgttggaacc
     6961 gctcttatca agaagtaccc aaagttggag tctgagttcg tttacggaga ttacaaggtt
     7021 tacgatgtta gaaagatgat cgctaagtct gagcaggaga tcggaaaggc taccgctaag
     7081 tacttcttct actctaacat catgaacttc ttcaagaccg agatcaccct tgctaacgga
     7141 gagatcagaa agagaccact tatcgagacc aacggagaga ccggagagat cgtttgggat
     7201 aagggaagag atttcgctac cgttagaaag gttctttcta tgccacaggt taacatcgtt
     7261 aagaaaaccg aggttcagac cggaggattc tctaaggagt ctatccttcc aaagagaaac
     7321 tctgataagt tgatcgctag aaagaaggat tgggacccaa agaagtacgg aggattcgat
     7381 tctccaaccg ttgcttactc tgttcttgtt gttgctaagg ttgagaaggg aaagtctaag
     7441 aagttgaagt ctgttaagga gcttcttgga atcaccatca tggagcgttc ttctttcgag
     7501 aagaacccaa tcgatttcct tgaggctaag ggatacaagg aggttaagaa ggatcttatc
     7561 atcaagttgc caaagtactc tcttttcgag cttgagaacg gaagaaagag aatgcttgct
     7621 tctgctggag agcttcagaa gggaaacgag cttgctcttc catctaagta cgttaacttc
     7681 ctttaccttg cttctcacta cgagaagttg aagggatctc cagaggataa cgagcagaag
     7741 cagcttttcg ttgagcagca caagcactac cttgatgaga tcatcgagca aatctctgag
     7801 ttctctaaga gagttatcct tgctgatgct aaccttgata aggttctttc tgcttacaac
     7861 aagcacagag ataagccaat cagagagcag gctgagaaca tcatccacct tttcaccctt
     7921 accaaccttg gtgctccagc tgctttcaag tacttcgata ccaccatcga tagaaaaaga
     7981 tacacctcta ccaaggaggt tcttgatgct acccttatcc accagtctat caccggactt
     8041 tacgagacca gaatcgatct ttctcagctt ggaggagata agagaccagc tgctaccaag
     8101 aaggctggac aggctaagaa gaagaagtga gacgtccgat cgttcaaaca tttggcaata
     8161 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     8221 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     8281 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     8341 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattgat
     8401 cccccctcga cagcttccgg aaagggcgaa ttcgcaactt tgtatacaaa agttgccgag
     8461 ctcgcggccg catgttgttg ttaccagaaa gtaaataaat gttcaatctc tgatgttctc
     8521 aagtaagtga gttttattgg gaataatatt aacttatgtt cttcttgcat ttgatttctt
     8581 tgccgctctc ttcttctatc ttaaatctgt gtatactatt tcactattgg gctttttatt
     8641 agtctataat gggactcaaa ataaggcttt ggcccacatc aaaaagataa gtcacaaatc
     8701 aaaactaaat tcagagtctt ttctcccaca tcggtcactg tactcatttt gtgtttgttt
     8761 atatattaca cgaaccgatc tttggtacgc tttggtcctt tgacgtcaag ttttagagct
     8821 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     8881 ggtgcttttt ttctagaccc agctttcttg tacaaagttg gcattaccat ggcttcgttg
     8941 aacaacggaa actcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
     9001 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
     9061 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
     9121 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
     9181 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
     9241 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
     9301 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
     9361 gagtcgaagt agtgattgca gcatttccca aattaatagt tttagagcta gaaatagcaa
     9421 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     9481 tctagaccca gctttcttgt acaaagttgg cattactgca ggtcgacgac ccagctttct
     9541 tgtacaaagt ggttcgataa ttcttaatta actagttcta atctagagtc ctgctttaat
     9601 gagatatgcg agacgcctat gatcgcatga tatttgcttt caattctgtt gtgcacgttg
     9661 taaaaaacct gagcatgtgt agctcagatc cttaccgccg gtttcggttc attctaatga
     9721 atatatcacc cgttactatc gtatttttat gaataatatt ctccgttcaa tttactgatt
     9781 gtaccctact acttatatgt acaatattaa aatgaaaaca atatattgtg ctgaataggt
     9841 ttatagcgac atctatgata gagcgccaca ataacaaaca attgcgtttt attattacaa
     9901 atccaatttt aaaaaaagcg gcagaaccgg tcaaacctaa aagactgatt acataaatct
     9961 tattcaaatt tcaaaagtgc cccaggggct agtatctacg acacaccgag cggcgaacta
    10021 ataacgctca ctgaagggaa ctccggttcc ccgccggcgc gcatgggtga gattccttga
    10081 agttgagtat tggccgtccg ctctaccgaa agttacgggc accattcaac ccggtccagc
    10141 acggcggccg ggtaaccgac ttgctgcccc gagaattatg cagcattttt ttggtgtatg
    10201 tgggccccaa atgaagtgca ggtcaaacct tgacagtgac gacaaatcgt tgggcgggtc
    10261 cagggcgaat tttgcgacaa catgtcgagg ctcagcagga cctgcaggca tgcaagctag
    10321 tgaagaatcg atttgctacg tattgcccct gcaggtcccc agattagcct tttcaatttc
    10381 agaaagaatg ctaacccaca gatggttaga gaggcttacg cagcaggtct catcaagacg
    10441 atctacccga gcaataatct ccaggaaatc aaataccttc ccaagaaggt taaagatgca
    10501 gtcaaaagat tcaggactaa ctgcatcaag aacacagaga aagatatatt tctcaagatc
    10561 agaagtacta ttccagtatg gacgattcaa ggcttgcttc acaaaccaag gcaagtaata
    10621 gagattggag tctctaaaaa ggtagttccc actgaatcaa aggccatgga gtcaaagatt
    10681 caaatagagg acctaacaga actcgccgta aagactggcg aacagttcat acagagtctc
    10741 ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga cacacttgtc
    10801 tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga gacttttcaa
    10861 caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg tcactttatt
    10921 gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
    10981 gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg
    11041 agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat
    11101 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
    11161 atataaggaa gttcatttca tttggagaga acacggggga ctctagagga tccatgagcc
    11221 cagaacgacg cccggccgac atccgccgtg ccaccgaggc ggacatgccg gcggtctgca
    11281 ccatcgtcaa ccactacatc gagacaagca cggtcaactt ccgtaccgag ccgcaggaac
    11341 cgcaggagtg gacggacgac ctcgtccgtc tgcgggagcg ctatccctgg ctcgtcgccg
    11401 aggtggacgg cgaggtcgcc ggcatcgcct acgcgggccc ctggaaggca cgcaacgcct
    11461 acgactggac ggccgagtcg accgtgtacg tctccccccg ccaccagcgg acgggactgg
    11521 gctccacgct ctacacccac ctgctgaagt ccctggaggc acagggcttc aagagcgtgg
    11581 tcgctgtcat cgggctgccc aacgacccga gcgtgcgcat gcacgaggcg ctcggatatg
    11641 ccccccgcgg catgctgcgg gcggccggct tcaagcacgg gaactggcat gacgtgggtt
    11701 tctggcagct ggacttcagc ctgccggtac cgccccgtcc ggtcctgccc gtcaccgaga
    11761 tctgaacgga gtgcgcgtgg catcgcccga gttggagctg gtacgggaac tcatcgaact
    11821 caactggcat acccgcaatg gtgaggtgga accgcggcgg atcgcgtacg accgtgccca
    11881 ggagggggta ccgagctcga atttccccga tcgttcaaac atttggcaat aaagtttctt
    11941 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt
    12001 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat
    12061 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta
    12121 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattaa ttcactggcc
    12181 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
    12241 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
    12301 caacagttgc gcagcctgaa tggcgcccgc tcctttcgct ttcttccctt cctttctcgc
    12361 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
    12421 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
    12481 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
    12541 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
    12601 ataagggatt ttgccgattt cggaaccacc atcaaacagg attttcgcct gctggggcaa
    12661 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
    12721 ttgcccgtct cactggtgaa aagaaaaacc accccagtac attaaaaacg tccgcaatgt
    12781 gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag
    12841 ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc
    12901 agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc
    12961 gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg
    13021 agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca aatatcatct
    13081 ccctcgcaga gatccgaatt atcagccttc ttattcattt ctcgcttaac cgtgacaggc
    13141 tgtcgatctt gagaactatg ccgacataat aggaaatcgc tggataaagc cgctgaggaa
    13201 gctgagtggc gctatttctt tagaagtgaa cgttgacgat atcaactccc ctatccattg
    13261 ctcaccgaat ggtacaggtc ggggacccga agttccgact gtcggcctga tgcatccccg
    13321 gctgatcgac cccagatctg gggctgagaa agcccagtaa ggaaacaact gtaggttcga
    13381 gtcgcgagat cccccggaac caaaggaagt aggttaaacc cgctccgatc aggccgagcc
    13441 acgccaggcc gagaacattg gttcctgtag gcatcgggat tggcggatca aacactaaag
    13501 ctactggaac gagcagaagt cctccggccg ccagttgcca ggcggtaaag gtgagcagag
    13561 gcacgggagg ttgccacttg cgggtcagca cggttccgaa cgccatggaa accgcccccg
    13621 ccaggcccgc tgcgacgccg acaggatcta gcgctgcgtt tggtgtcaac accaacagcg
    13681 ccacgcccgc agttccgcaa atagccccca ggaccgccat caatcgtatc gggctaccta
    13741 gcagagcggc agagatgaac acgaccatca gcggctgcac agcgcctacc gtcgccgcga
    13801 ccccgcccgg caggcggtag accgaaataa acaacaagct ccagaatagc gaaatattaa
    13861 gtgcgccgag gatgaagatg cgcatccacc agattcccgt tggaatctgt cggacgatca
    13921 tcacgagcaa taaacccgcc ggcaacgccc gcagcagcat accggcgacc cctcggcctc
    13981 gctgttcggg ctccacgaaa acgccggaca gatgcgcctt gtgagcgtcc ttggggccgt
    14041 cctcctgttt gaagaccgac agcccaatga tctcgccgtc gatgtaggcg ccgaatgcca
    14101 cggcatctcg caaccgttca gcgaacgcct ccatgggctt tttctcctcg tgctcgtaaa
    14161 cggacccgaa catctctgga gctttcttca gggccgacaa tcggatctcg cggaaatcct
    14221 gcacgtcggc cgctccaagc cgtcgaatct gagccttaat cacaattgtc aattttaatc
    14281 ctctgtttat cggcagttcg tagagcgcgc cgtgcgtccc gagcgatact gagcgaagca
    14341 agtgcgtcga gcagtgcccg cttgttcctg aaatgccagt aaagcgctgg ctgctgaacc
    14401 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
    14461 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
    14521 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
    14581 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
    14641 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
    14701 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
    14761 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
    14821 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
    14881 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
    14941 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
    15001 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
    15061 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
    15121 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
    15181 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct cgctgacctg
    15241 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
    15301 gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agacgacgcg aacgctccac
    15361 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
    15421 ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
    15481 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
    15541 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
    15601 gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg ccttggtgtc
    15661 cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc cgtccttctc
    15721 gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc ccaaatactt
    15781 gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg tgcgctcctg
    15841 cttgtcgccg gcatcgttgc gccacatcta ggtactaaaa caattcatcc agtaaaatat
    15901 aatattttat tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat
    15961 actgttcttc cccgatatcc tccctgatcg accggacgca gaaggcaatg tcataccact
    16021 tgtccgccct gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa
    16081 agatgttgct gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg
    16141 tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg agtgtcttct tcccagtttt
    16201 cgcaatccac atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt
    16261 ctaagctatt cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact
    16321 ccgcatacag ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa
    16381 ggacgccatc ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca
    16441 ggacctttgg aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca
    16501 catcataggt ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt
    16561 ctcccaccag cttatatacc ttagcaggag acattccttc cgtatctttt acgcagcggt
    16621 atttttcgat cagttttttc aattccggtg atattctcat tttagccatt tattatttcc
    16681 ttcctctttt ctacagtatt taaagatacc ccaagaagct aattataaca agacgaactc
    16741 caattcactg ttccttgcat tctaaaacct taaataccag aaaacagctt tttcaaagtt
    16801 gttttcaaag ttggcgtata acatagtatc gacggagccg attttgaaac cacaattatg
    16861 ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg ctccagtggc
    16921 ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc gtaacggcaa
    16981 aagcaccgcc ggacatcagc gctatctctg ctctcactgc cgtaaaacat ggcaactgca
    17041 gttcacttac accgcttctc aacccggtac gcaccagaaa atcattgata tggccatgaa
    17101 tggcgttgga tgccgggcaa cagcccgcat tatgggcgtt ggcctcaaca cgattttacg
    17161 tcacttaaaa aactcaggcc gcagtcggta acctcgcgca tacagccggg cagtgacgtc
    17221 atcgtctgcg cggaaatgga cgaacagtgg ggctatgtcg gggctaaatc gcgccagcgc
    17281 tggctgtttt acgcgtatga cagtctccgg aagacggttg ttgcgcacgt attcggtgaa
    17341 cgcactatgg cgacgctggg gcgtcttatg agcctgctgt caccctttga cgtggtgata
    17401 tggatgacgg atggctggcc gctgtatgaa tcccgcctga agggaaagct gcacgtaatc
    17461 agcaagcgat atacgcagcg aattgagcgg cataacctga atctgaggca gcacctggca
    17521 cggctgggac ggaagtcgct gtcgttctca aaatcggtgg agctgcatga caaagtcatc
    17581 gggcattatc tgaacataaa acactatcaa taagttggag tcattaccca attatgatag
    17641 aatttacaag ctataaggtt attgtcctgg gtttcaagca ttagtccatg caagttttta
    17701 tgctttgccc attctataga tatattgata agcgcgctgc ctatgccttg ccccctgaaa
    17761 tccttacata cggcgatatc ttctatataa aagatatatt atcttatcag tattgtcaat
    17821 atattcaagg caatctgcct cctcatcctc ttcatcctct tcgtcttggt agctttttaa
    17881 atatggcgct tcatagagta attctgtaaa ggtccaattc tcgttttcat acctcggtat
    17941 aatcttacct atcacctcaa atggttcgct gggtttatcg cacccccgaa cacgagcacg
    18001 gcacccgcga ccactatgcc aagaatgccc aaggtaaaaa ttgccggccc cgccatgaag
    18061 tccgtgaatg ccccgacggc cgaagtgaag ggcaggccgc cacccaggcc gccgccctca
    18121 ctgcccggca cctggtcgct gaatgtcgat gccagcacct gcggcacgtc aatgcttccg
    18181 ggcgtcgcgc tcgggctgat cgcccatccc gttactgccc cgatcccggc aatggcaagg
    18241 actgccagcg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg
    18301 ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt
    18361 cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat
    18421 tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt
    18481 gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca
    18541 tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc
    18601 gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg
    18661 gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc
    18721 cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca
    18781 agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca
    18841 acgccggcgg ccggccgcgg tgtctcgcac acggcttcga cggcgtttct ggcgcgtttg
    18901 cagggccata gacggccgcc agcccagcgg cgagggcaac cagcccggtg agcgtcggaa
    18961 aggcgctcgg tcttgccttg ctcgtcggt
//
