LOCUS       sequence-202693-       18337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g020630 (FmoI). Also 
            expresses Cas9 from AtUBQ10 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18337)
  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 18337)
  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: pSC20
FEATURES             Location/Qualifiers
     source          1..18337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             49..96
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             103..123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(148..1584,1774..4437)
                     /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          1585..1773
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4438..4485
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4496..4748
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4793..4813
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5168..5243
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5255..5279)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5757..5832
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5844..5868)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5887..5911)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      5975..6227
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6245..6261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6245..6261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6527..6551
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7016..7666
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7854..7871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8105..8124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8347..8456
                     /note="incP origin of transfer"
     CDS             8489..8860
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      8987..9702
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10325..11092
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11284..12078
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12377..13525
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    15121..15145
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15392..15413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15604..15623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15763..15779
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15792..16044)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16188..16739)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(16760..17105)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16797..16817)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17641..17657
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaacaagttt gtacaaaaaa gcaggctccg aattcgccct tcaccatgga ttacaaggat
       61 gatgatgata aggattacaa ggatgatgat gataagatgg ctccaaagaa gaagagaaag
      121 gttggaatcc acggagttcc agctgctgat aagaagtact ctatcggact tgacatcgga
      181 accaactctg ttggatgggc tgttatcacc gatgagtaca aggttccatc taagaagttc
      241 aaggttcttg gaaacaccga tagacactct atcaagaaga accttatcgg tgctcttctt
      301 ttcgattctg gagagaccgc tgaggctacc agattgaaga gaaccgctag aagaagatac
      361 accagaagaa agaacagaat ctgctacctt caggaaatct tctctaacga gatggctaag
      421 gttgatgatt ctttcttcca cagacttgag gagtctttcc ttgttgagga ggataagaag
      481 cacgagagac acccaatctt cggaaacatc gttgatgagg ttgcttacca cgagaagtac
      541 ccaaccatct accaccttag aaagaagttg gttgattcta ccgataaggc tgatcttaga
      601 cttatctacc ttgctcttgc tcacatgatc aagttcagag gacacttcct tatcgaggga
      661 gaccttaacc cagataactc tgatgttgat aagttgttca tccagcttgt tcagacctac
      721 aaccagcttt tcgaggagaa cccaatcaac gcttctggag ttgatgctaa ggctatcctt
      781 tctgctagac tttctaagtc tcgtagactt gagaacctta tcgctcagct tccaggagag
      841 aagaagaacg gacttttcgg aaaccttatc gctctttctc ttggacttac cccaaacttc
      901 aagtctaact tcgatcttgc tgaggatgct aagttgcagc tttctaagga tacctacgat
      961 gatgatcttg ataaccttct tgctcagatc ggagatcagt acgctgatct tttccttgct
     1021 gctaagaacc tttctgatgc tatccttctt tctgacatcc ttagagttaa caccgagatc
     1081 accaaggctc cactttctgc ttctatgatc aagagatacg atgagcacca ccaggatctt
     1141 acccttttga aggctcttgt tagacagcag cttccagaga agtacaagga aatcttcttc
     1201 gatcagtcta agaacggata cgctggatac atcgatggag gagcttctca ggaggagttc
     1261 tacaagttca tcaagccaat ccttgagaag atggatggaa ccgaggagct tcttgttaag
     1321 ttgaacagag aggatcttct tagaaagcag agaaccttcg ataacggatc tatcccacac
     1381 cagatccacc ttggagagct tcacgctatc cttcgtagac aggaggattt ctacccattc
     1441 ttgaaggata acagagagaa gatcgagaag atccttacct tcagaatccc atactacgtt
     1501 ggaccacttg ctagaggaaa ctctcgtttc gcttggatga ccagaaagtc tgaggagacc
     1561 atcacccctt ggaacttcga ggaggtaagt ttctgcttct acctttgata tatatataat
     1621 aattatcatt aattagtagt aatataatat ttcaaatatt tttttcaaaa taaaagaatg
     1681 tagtatatag caattgcttt tctgtagttt ataagtgtgt atattttaat ttataacttt
     1741 tctaatatat gaccaaaatt tgttgatgtg caggttgttg ataagggagc ttctgctcag
     1801 tctttcatcg agagaatgac caacttcgat aagaaccttc caaacgagaa ggttcttcca
     1861 aagcactctc ttctttacga gtacttcacc gtttacaacg agcttaccaa ggttaagtac
     1921 gttaccgagg gaatgagaaa gccagctttc ctttctggag agcagaagaa ggctatcgtt
     1981 gatcttcttt tcaagaccaa cagaaaggtt accgttaagc agttgaagga ggattacttc
     2041 aagaagatcg agtgcttcga ttctgttgaa atctctggag ttgaggatag attcaacgct
     2101 tctcttggaa cctaccacga tcttttgaag atcatcaagg ataaggattt ccttgataac
     2161 gaggagaacg aggacatcct tgaggacatc gttcttaccc ttaccctttt cgaggataga
     2221 gagatgatcg aggagagact caagacctac gctcaccttt tcgatgataa ggttatgaag
     2281 cagttgaaga gaagaagata caccggatgg ggtagacttt ctcgtaagtt gatcaacgga
     2341 atcagagata agcagtctgg aaagaccatc cttgatttct tgaagtctga tggattcgct
     2401 aacagaaact tcatgcagct tatccacgat gattctctta ccttcaagga ggacatccag
     2461 aaggctcagg tttctggaca gggagattct cttcacgagc acatcgctaa ccttgctgga
     2521 tctccagcta tcaagaaggg aatccttcag accgttaagg ttgttgatga gcttgttaag
     2581 gttatgggta gacacaagcc agagaacatc gttatcgaga tggctagaga gaaccagacc
     2641 acccagaagg gacagaagaa ctctcgtgag agaatgaaga gaatcgagga gggaatcaag
     2701 gagcttggat ctcaaatctt gaaggagcac ccagttgaga acacccagct tcagaacgag
     2761 aagttgtacc tttactacct tcagaacgga agagatatgt acgttgatca ggagcttgac
     2821 atcaacagac tttctgatta cgatgttgat cacatcgttc cacagtcttt cttgaaggat
     2881 gattctatcg ataacaaggt tcttacccgt tctgataaga acagaggaaa gtctgataac
     2941 gttccatctg aggaggttgt taagaagatg aagaactact ggagacagct tcttaacgct
     3001 aagttgatca cccagagaaa gttcgataac cttaccaagg ctgagagagg aggactttct
     3061 gagcttgata aggctggatt catcaagaga cagcttgttg agaccagaca gatcaccaag
     3121 cacgttgctc agatccttga ttctcgtatg aacaccaagt acgatgagaa cgataagttg
     3181 atcagagagg ttaaggttat caccttgaag tctaagttgg tttctgattt cagaaaggat
     3241 ttccagttct acaaggttag agagatcaac aactaccacc acgctcacga tgcttacctt
     3301 aacgctgttg ttggaaccgc tcttatcaag aagtacccaa agttggagtc tgagttcgtt
     3361 tacggagatt acaaggttta cgatgttaga aagatgatcg ctaagtctga gcaggagatc
     3421 ggaaaggcta ccgctaagta cttcttctac tctaacatca tgaacttctt caagaccgag
     3481 atcacccttg ctaacggaga gatcagaaag agaccactta tcgagaccaa cggagagacc
     3541 ggagagatcg tttgggataa gggaagagat ttcgctaccg ttagaaaggt tctttctatg
     3601 ccacaggtta acatcgttaa gaaaaccgag gttcagaccg gaggattctc taaggagtct
     3661 atccttccaa agagaaactc tgataagttg atcgctagaa agaaggattg ggacccaaag
     3721 aagtacggag gattcgattc tccaaccgtt gcttactctg ttcttgttgt tgctaaggtt
     3781 gagaagggaa agtctaagaa gttgaagtct gttaaggagc ttcttggaat caccatcatg
     3841 gagcgttctt ctttcgagaa gaacccaatc gatttccttg aggctaaggg atacaaggag
     3901 gttaagaagg atcttatcat caagttgcca aagtactctc ttttcgagct tgagaacgga
     3961 agaaagagaa tgcttgcttc tgctggagag cttcagaagg gaaacgagct tgctcttcca
     4021 tctaagtacg ttaacttcct ttaccttgct tctcactacg agaagttgaa gggatctcca
     4081 gaggataacg agcagaagca gcttttcgtt gagcagcaca agcactacct tgatgagatc
     4141 atcgagcaaa tctctgagtt ctctaagaga gttatccttg ctgatgctaa ccttgataag
     4201 gttctttctg cttacaacaa gcacagagat aagccaatca gagagcaggc tgagaacatc
     4261 atccaccttt tcacccttac caaccttggt gctccagctg ctttcaagta cttcgatacc
     4321 accatcgata gaaaaagata cacctctacc aaggaggttc ttgatgctac ccttatccac
     4381 cagtctatca ccggacttta cgagaccaga atcgatcttt ctcagcttgg aggagataag
     4441 agaccagctg ctaccaagaa ggctggacag gctaagaaga agaagtgaga cgtccgatcg
     4501 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     4561 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     4621 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     4681 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     4741 actagatcgg gaattgatcc cccctcgaca gcttccggaa agggcgaatt cgcaactttg
     4801 tatacaaaag ttgccgagct cgcggccgca tgttgttgtt accagaaagt aaataaatgt
     4861 tcaatctctg atgttctcaa gtaagtgagt tttattggga ataatattaa cttatgttct
     4921 tcttgcattt gatttctttg ccgctctctt cttctatctt aaatctgtgt atactatttc
     4981 actattgggc tttttattag tctataatgg gactcaaaat aaggctttgg cccacatcaa
     5041 aaagataagt cacaaatcaa aactaaattc agagtctttt ctcccacatc ggtcactgta
     5101 ctcattttgt gtttgtttat atattacacg aaccgatctt tggtacggaa gccactgatt
     5161 caattgggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     5221 aaaaagtggc accgagtcgg tgcttttttt ctagacccag ctttcttgta caaagttggc
     5281 attaccatgg cttcgttgaa caacggaaac tcgacttgcc ttccgcacaa tacatcattt
     5341 cttcttagct ttttttcttc ttcttcgttc atacagtttt tttttgttta tcagcttaca
     5401 ttttcttgaa ccgtagcttt cgttttcttc tttttaactt tccattcgga gtttttgtat
     5461 cttgtttcat agtttgtccc aggattagaa tgattaggca tcgaaccttc aagaatttga
     5521 ttgaataaaa catcttcatt cttaagatat gaagataatc ttcaaaaggc ccctgggaat
     5581 ctgaaagaag agaagcaggc ccatttatat gggaaagaac aatagtattt cttatatagg
     5641 cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg cttagataag aaaacgaagc
     5701 tgagtttata tacagctaga gtcgaagtag tgattgggat gcttacctcc tgaccagttt
     5761 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     5821 ccgagtcggt gctttttttc tagacccagc tttcttgtac aaagttggca ttactgcagg
     5881 tcgacgaccc agctttcttg tacaaagtgg ttcgataatt cttaattaac tagttctaga
     5941 gcggccgcca ccgcggtgga gctcgaattt ccccgatcgt tcaaacattt ggcaataaag
     6001 tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa
     6061 ttacgttaag catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt
     6121 tatgattaga gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc
     6181 aaactaggat aaattatcgc gcgcggtgtc atctatgtta ctagatcggg aattcgtaat
     6241 catggtcata gctgtttcct gcctagtcgg agcggccgcg tccgtctagc tgatagtgac
     6301 cttaggcgac ttttgaacgc gcaataatgg tttctgacgt atgtgcttag ctcattaaac
     6361 tccagaaacc cgcggctgag tggctccttc aacgttgcgg ttctgtcagt tccaaacgta
     6421 aaacggcttg tcccgcgtca tcggcggggg tcataacgtg actcccttaa ttctccgctc
     6481 atgatcagat tgtcgtttcc cgccttcagt ttaaactatc agtgtttgac aggatatatt
     6541 ggcgggtaaa cctaagagaa aagagcgttt attagaataa tcggatattt aaaagggcgt
     6601 gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt tccccagatc
     6661 tggcgccggc cagcgagacg agcaagattg gccgccgccc gaaacgatcc gacagcgcgc
     6721 ccagcacagg tgcgcaggca aattgcacca acgcatacag cgccagcaga atgccatagt
     6781 gggcggtgac gtcgttcgag tgaaccagat cgcgcaggag gcccggcagc accggcataa
     6841 tcaggccgat gccgacagcg tcgagcgcga cagtgctcag aattacgatc aggggtatgt
     6901 tgggtttcac gtctggcctc cggaccagcc tccgctggtc cgattgaacg cgcggattct
     6961 ttatcactga taagttggtg gacatattat gtttatcagt gataaagtgt caagcatgac
     7021 aaagttgcag ccgaatacag tgatccgtgc cgccctggac ctgttgaacg aggtcggcgt
     7081 agacggtctg acgacacgca aactggcgga acggttgggg gttcagcagc cggcgcttta
     7141 ctggcacttc aggaacaagc gggcgctgct cgacgcactg gccgaagcca tgctggcgga
     7201 gaatcatacg cattcggtgc cgagagccga cgacgactgg cgctcatttc tgatcgggaa
     7261 tgcccgcagc ttcaggcagg cgctgctcgc ctaccgcgat ggcgcgcgca tccatgccgg
     7321 cacgcgaccg ggcgcaccgc agatggaaac ggccgacgcg cagcttcgct tcctctgcga
     7381 ggcgggtttt tcggccgggg acgccgtcaa tgcgctgatg acaatcagct acttcactgt
     7441 tggggccgtg cttgaggagc aggccggcga cagcgatgcc ggcgagcgcg gcggcaccgt
     7501 tgaacaggct ccgctctcgc cgctgttgcg ggccgcgata gacgccttcg acgaagccgg
     7561 tccggacgca gcgttcgagc agggactcgc ggtgattgtc gatggattgg cgaaaaggag
     7621 gctcgttgtc aggaacgttg aaggaccgag aaagggtgac gattgatcag gaccgctgcc
     7681 ggagcgcaac ccactcacta cagcagagcc atgtagacaa catcccctcc ccctttccac
     7741 cgcgtcagac gcccgtagca gcccgctacg ggctttttca tgccctgccc tagcgtccaa
     7801 gcctcacggc cgcgctcggc ctctctggcg gccttctggc gctcttccgc ttcctcgctc
     7861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     8041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     8101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     8161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gcttttccgc
     8221 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc
     8281 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg
     8341 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt
     8401 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccggctac
     8461 cgccggcgta acagatgagg gcaagcggat ggctgatgaa accaagccaa ccaggaaggg
     8521 cagcccacct atcaaggtgt actgccttcc agacgaacga agagcgattg aggaaaaggc
     8581 ggcggcggcc ggcatgagcc tgtcggccta cctgctggcc gtcggccagg gctacaaaat
     8641 cacgggcgtc gtggactatg agcacgtccg cgagctggcc cgcatcaatg gcgacctggg
     8701 ccgcctgggc ggcctgctga aactctggct caccgacgac ccgcgcacgg cgcggttcgg
     8761 tgatgccacg atcctcgccc tgctggcgaa gatcgaagag aagcaggacg agcttggcaa
     8821 ggtcatgatg ggcgtggtcc gcccgagggc agagccatga cttttttagc cgctaaaacg
     8881 gccggggggt gcgcgtgatt gccaagcacg tccccatgcg ctccatcaag aagagcgact
     8941 tcgcggagct ggtgaagtac atcaccgacg agcaaggcaa gaccgagcgc ctttccgacg
     9001 ctcaccgggc tggttgccct cgccgctggg ctggcggccg tctatggccc tgcaaacgcg
     9061 ccagaaacgc cgtcgaagcc gtgtgcgaga caccgcggcc ggccgccggc gttgtggata
     9121 cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca cttgaggggc
     9181 cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc cggcgacgtg
     9241 gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat
     9301 gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg gcgcgactac
     9361 tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca gatgaggggc
     9421 gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt gcaagggttt
     9481 ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac caatatttat
     9541 aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg
     9601 tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc ccccaggggc
     9661 tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agcgctggca gtccttgcca
     9721 ttgccgggat cggggcagta acgggatggg cgatcagccc gagcgcgacg cccggaagca
     9781 ttgacgtgcc gcaggtgctg gcatcgacat tcagcgacca ggtgccgggc agtgagggcg
     9841 gcggcctggg tggcggcctg cccttcactt cggccgtcgg ggcattcacg gacttcatgg
     9901 cggggccggc aatttttacc ttgggcattc ttggcatagt ggtcgcgggt gccgtgctcg
     9961 tgttcggggg tgcgataaac ccagcgaacc atttgaggtg ataggtaaga ttataccgag
    10021 gtatgaaaac gagaattgga cctttacaga attactctat gaagcgccat atttaaaaag
    10081 ctaccaagac gaagaggatg aagaggatga ggaggcagat tgccttgaat atattgacaa
    10141 tactgataag ataatatatc ttttatatag aagatatcgc cgtatgtaag gatttcaggg
    10201 ggcaaggcat aggcagcgcg cttatcaata tatctataga atgggcaaag cataaaaact
    10261 tgcatggact aatgcttgaa acccaggaca ataaccttat agcttgtaaa ttctatcata
    10321 attgggtaat gactccaact tattgatagt gttttatgtt cagataatgc ccgatgactt
    10381 tgtcatgcag ctccaccgat tttgagaacg acagcgactt ccgtcccagc cgtgccaggt
    10441 gctgcctcag attcaggtta tgccgctcaa ttcgctgcgt atatcgcttg ctgattacgt
    10501 gcagctttcc cttcaggcgg gattcataca gcggccagcc atccgtcatc catatcacca
    10561 cgtcaaaggg tgacagcagg ctcataagac gccccagcgt cgccatagtg cgttcaccga
    10621 atacgtgcgc aacaaccgtc ttccggagac tgtcatacgc gtaaaacagc cagcgctggc
    10681 gcgatttagc cccgacatag ccccactgtt cgtccatttc cgcgcagacg atgacgtcac
    10741 tgcccggctg tatgcgcgag gttaccgact gcggcctgag ttttttaagt gacgtaaaat
    10801 cgtgttgagg ccaacgccca taatgcgggc tgttgcccgg catccaacgc cattcatggc
    10861 catatcaatg attttctggt gcgtaccggg ttgagaagcg gtgtaagtga actgcagttg
    10921 ccatgtttta cggcagtgag agcagagata gcgctgatgt ccggcggtgc ttttgccgtt
    10981 acgcaccacc ccgtcagtag ctgaacagga gggacagctg atagaaacag aagccactgg
    11041 agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca cccataattg
    11101 tggtttcaaa atcggctccg tcgatactat gttatacgcc aactttgaaa acaactttga
    11161 aaaagctgtt ttctggtatt taaggtttta gaatgcaagg aacagtgaat tggagttcgt
    11221 cttgttataa ttagcttctt ggggtatctt taaatactgt agaaaagagg aaggaaataa
    11281 taaatggcta aaatgagaat atcaccggaa ttgaaaaaac tgatcgaaaa ataccgctgc
    11341 gtaaaagata cggaaggaat gtctcctgct aaggtatata agctggtggg agaaaatgaa
    11401 aacctatatt taaaaatgac ggacagccgg tataaaggga ccacctatga tgtggaacgg
    11461 gaaaaggaca tgatgctatg gctggaagga aagctgcctg ttccaaaggt cctgcacttt
    11521 gaacggcatg atggctggag caatctgctc atgagtgagg ccgatggcgt cctttgctcg
    11581 gaagagtatg aagatgaaca aagccctgaa aagattatcg agctgtatgc ggagtgcatc
    11641 aggctctttc actccatcga catatcggat tgtccctata cgaatagctt agacagccgc
    11701 ttagccgaat tggattactt actgaataac gatctggccg atgtggattg cgaaaactgg
    11761 gaagaagaca ctccatttaa agatccgcgc gagctgtatg attttttaaa gacggaaaag
    11821 cccgaagagg aacttgtctt ttcccacggc gacctgggag acagcaacat ctttgtgaaa
    11881 gatggcaaag taagtggctt tattgatctt gggagaagcg gcagggcgga caagtggtat
    11941 gacattgcct tctgcgtccg gtcgatcagg gaggatatcg gggaagaaca gtatgtcgag
    12001 ctattttttg acttactggg gatcaagcct gattgggaga aaataaaata ttatatttta
    12061 ctggatgaat tgttttagta cctagatgtg gcgcaacgat gccggcgaca agcaggagcg
    12121 caccgacttc ttccgcatca agtgttttgg ctctcaggcc gaggcccacg gcaagtattt
    12181 gggcaagggg tcgctggtat tcgtgcaggg caagattcgg aataccaagt acgagaagga
    12241 cggccagacg gtctacggga ccgacttcat tgccgataag gtggattatc tggacaccaa
    12301 ggcaccaggc gggtcaaatc aggaataagg gcacattgcc ccggcgtgag tcggggcaat
    12361 cccgcaagga gggtgaatga atcggacgtt tgaccggaag gcatacaggc aagaactgat
    12421 cgacgcgggg ttttccgccg aggatgccga aaccatcgca agccgcaccg tcatgcgtgc
    12481 gccccgcgaa accttccagt ccgtcggctc gatggtccag caagctacgg ccaagatcga
    12541 gcgcgacagc gtgcaactgg ctccccctgc cctgcccgcg ccatcggccg ccgtggagcg
    12601 ttcgcgtcgt ctcgaacagg aggcggcagg tttggcgaag tcgatgacca tcgacacgcg
    12661 aggaactatg acgaccaaga agcgaaaaac cgccggcgag gacctggcaa aacaggtcag
    12721 cgaggccaag caggccgcgt tgctgaaaca cacgaagcag cagatcaagg aaatgcagct
    12781 ttccttgttc gatattgcgc cgtggccgga cacgatgcga gcgatgccaa acgacacggc
    12841 ccgctctgcc ctgttcacca cgcgcaacaa gaaaatcccg cgcgaggcgc tgcaaaacaa
    12901 ggtcattttc cacgtcaaca aggacgtgaa gatcacctac accggcgtcg agctgcgggc
    12961 cgacgatgac gaactggtgt ggcagcaggt gttggagtac gcgaagcgca cccctatcgg
    13021 cgagccgatc accttcacgt tctacgagct ttgccaggac ctgggctggt cgatcaatgg
    13081 ccggtattac acgaaggccg aggaatgcct gtcgcgccta caggcgacgg cgatgggctt
    13141 cacgtccgac cgcgttgggc acctggaatc ggtgtcgctg ctgcaccgct tccgcgtcct
    13201 ggaccgtggc aagaaaacgt cccgttgcca ggtcctgatc gacgaggaaa tcgtcgtgct
    13261 gtttgctggc gaccactaca cgaaattcat atgggagaag taccgcaagc tgtcgccgac
    13321 ggcccgacgg atgttcgact atttcagctc gcaccgggag ccgtacccgc tcaagctgga
    13381 aaccttccgc ctcatgtgcg gatcggattc cacccgcgtg aagaagtggc gcgagcaggt
    13441 cggcgaagcc tgcgaagagt tgcgaggcag cggcctggtg gaacacgcct gggtcaatga
    13501 tgacctggtg cattgcaaac gctagggcct tgtggggtca gttccggctg ggggttcagc
    13561 agccagcgct ttactggcat ttcaggaaca agcgggcact gctcgacgca cttgcttcgc
    13621 tcagtatcgc tcgggacgca cggcgcgctc tacgaactgc cgataaacag aggattaaaa
    13681 ttgacaattg tgattaaggc tcagattcga cggcttggag cggccgacgt gcaggatttc
    13741 cgcgagatcc gattgtcggc cctgaagaaa gctccagaga tgttcgggtc cgtttacgag
    13801 cacgaggaga aaaagcccat ggaggcgttc gctgaacggt tgcgagatgc cgtggcattc
    13861 ggcgcctaca tcgacggcga gatcattggg ctgtcggtct tcaaacagga ggacggcccc
    13921 aaggacgctc acaaggcgca tctgtccggc gttttcgtgg agcccgaaca gcgaggccga
    13981 ggggtcgccg gtatgctgct gcgggcgttg ccggcgggtt tattgctcgt gatgatcgtc
    14041 cgacagattc caacgggaat ctggtggatg cgcatcttca tcctcggcgc acttaatatt
    14101 tcgctattct ggagcttgtt gtttatttcg gtctaccgcc tgccgggcgg ggtcgcggcg
    14161 acggtaggcg ctgtgcagcc gctgatggtc gtgttcatct ctgccgctct gctaggtagc
    14221 ccgatacgat tgatggcggt cctgggggct atttgcggaa ctgcgggcgt ggcgctgttg
    14281 gtgttgacac caaacgcagc gctagatcct gtcggcgtcg cagcgggcct ggcgggggcg
    14341 gtttccatgg cgttcggaac cgtgctgacc cgcaagtggc aacctcccgt gcctctgctc
    14401 acctttaccg cctggcaact ggcggccgga ggacttctgc tcgttccagt agctttagtg
    14461 tttgatccgc caatcccgat gcctacagga accaatgttc tcggcctggc gtggctcggc
    14521 ctgatcggag cgggtttaac ctacttcctt tggttccggg ggatctcgcg actcgaacct
    14581 acagttgttt ccttactggg ctttctcagc cccagatctg gggtcgatca gccggggatg
    14641 catcaggccg acagtcggaa cttcgggtcc ccgacctgta ccattcggtg agcaatggat
    14701 aggggagttg atatcgtcaa cgttcacttc taaagaaata gcgccactca gcttcctcag
    14761 cggctttatc cagcgatttc ctattatgtc ggcatagttc tcaagatcga cagcctgtca
    14821 cggttaagcg agaaatgaat aagaaggctg ataattcgga tctctgcgag ggagatgata
    14881 tttgatcaca ggcagcaacg ctctgtcatc gttacaatca acatgctacc ctccgcgaga
    14941 tcatccgtgt ttcaaacccg gcagcttagt tgccgttctt ccgaatagca tcggtaacat
    15001 gagcaaagtc tgccgcctta caacggctct cccgctgacg ccgtcccgga ctgatgggct
    15061 gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg
    15121 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
    15181 gacgttttta atgtactggg gtggtttttc ttttcaccag tgagacgggc aacagctgat
    15241 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
    15301 gcaggcgaaa atcctgtttg atggtggttc cgaaatcggc aaaatccctt ataaatcaaa
    15361 agaatagccc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
    15421 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
    15481 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
    15541 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
    15601 ggaagggaag aaagcgaaag gagcgggcgc cattcaggct gcgcaactgt tgggaagggc
    15661 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
    15721 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
    15781 aattaattcc cgatctagta acatagatga caccgcgcgc gataatttat cctagtttgc
    15841 gcgctatatt ttgttttcta tcgcgtatta aatgtataat tgcgggactc taatcataaa
    15901 aacccatctc ataaataacg tcatgcatta catgttaatt attacatgct taacgtaatt
    15961 caacagaaat tatatgataa tcatcgcaag accggcaaca ggattcaatc ttaagaaact
    16021 ttattgccaa atgtttgaac gatcggggaa attcgagctc ggtaccccct cctgggcacg
    16081 gtcgtacgcg atccgccgcg gttccacctc accattgcgg gtatgccagt tgagttcgat
    16141 gagttcccgt accagctcca actcgggcga tgccacgcgc actccgttca gatctcggtg
    16201 acgggcagga ccggacgggg cggtaccggc aggctgaagt ccagctgcca gaaacccacg
    16261 tcatgccagt tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg ggcatatccg
    16321 agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc gaccacgctc
    16381 ttgaagccct gtgcctccag ggacttcagc aggtgggtgt agagcgtgga gcccagtccc
    16441 gtccgctggt ggcgggggga gacgtacacg gtcgactcgg ccgtccagtc gtaggcgttg
    16501 cgtgccttcc aggggcccgc gtaggcgatg ccggcgacct cgccgtccac ctcggcgacg
    16561 agccagggat agcgctcccg cagacggacg aggtcgtccg tccactcctg cggttcctgc
    16621 ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat ggtgcagacc
    16681 gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc tgggctcatg
    16741 gatcctctag agtcccccgt gttctctcca aatgaaatga acttccttat atagaggaag
    16801 ggtcttgcga aggatagtgg gattgtgcgt catcccttac gtcagtggag atatcacatc
    16861 aatccacttg ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg
    16921 gtgggggtcc atctttggga ccactgtcgg cagaggcatc ttcaacgatg gcctttcctt
    16981 tatcgcaatg atggcatttg taggagccac cttccttttc cactatcttc acaataaagt
    17041 gacagatagc tgggcaatgg aatccgagga ggtttccgga tattaccctt tgttgaaaag
    17101 tctcaattgc cctttggtct tctgagactg tatctttgat atttttggag tagacaagtg
    17161 tgtcgtgctc caccatgttg acgaagattt tcttcttgtc attgagtcgt aagagactct
    17221 gtatgaactg ttcgccagtc tttacggcga gttctgttag gtcctctatt tgaatctttg
    17281 actccatggc ctttgattca gtgggaacta cctttttaga gactccaatc tctattactt
    17341 gccttggttt gtgaagcaag ccttgaatcg tccatactgg aatagtactt ctgatcttga
    17401 gaaatatatc tttctctgtg ttcttgatgc agttagtcct gaatcttttg actgcatctt
    17461 taaccttctt gggaaggtat ttgatttcct ggagattatt gctcgggtag atcgtcttga
    17521 tgagacctgc tgcgtaagcc tctctaacca tctgtgggtt agcattcttt ctgaaattga
    17581 aaaggctaat ctggggacct gcaggggcaa taccgccagg gttttcccag tcacgacgtt
    17641 gtaaaacgac ggccagtgcc aagctttcga cgagtcagta ataaacggcg tcaaagtggt
    17701 tgcagccggc acacacgagt cgtgtttatc aactcaaagc acaaatactt ttcctcaacc
    17761 taaaaataag gcaattagcc aaaaacaact ttgcgtgtaa acaacgctca atacacgtgt
    17821 cattttatta ttagctattg cttcaccgcc ttagctttct cgtgacctag tcgtcctcgt
    17881 cttttcttct tcttcttcta taaaacaata cccaaagagc tcttcttctt cacaattcag
    17941 atttcaattt ctcaaaatct taaaaacttt ctctcaattc tctctaccgt gatcaaggta
    18001 aatttctgtg ttccttattc tctcaaaatc ttcgattttg ttttcgttcg atcccaattt
    18061 cgtatatgtt ctttggttta gattctgtta atcttagatc gaagacgatt ttctgggttt
    18121 gatcgttaga tatcatctta attctcgatt agggtttcat agatatcatc cgatttgttc
    18181 aaataatttg agttttgtcg aataattact cttcgatttg tgatttctat ctagatctgg
    18241 tgttagtttc tagtttgtgc gatcgaattt gtagattaat ctgagttttt ctgattaaca
    18301 gggtaccggg ccccccctcg aggcgcgcca agctatc
//
