LOCUS       sequence-202698-       18379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101120, Medtr2g101130 
            (Acre1). Also expresses Cas9 from Gmubi promoter from AtUBQ10 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18379)
  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 18379)
  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: pSC22
FEATURES             Location/Qualifiers
     source          1..18379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    63..87
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             109..156
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             163..183
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(208..1644,1834..4497)
                     /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          1645..1833
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4498..4545
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4556..4808
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4853..4873
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5228..5303
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5315..5339)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5816..5891
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5903..5927)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5946..5970)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5989..6013)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6077..6329
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6347..6363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6347..6363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6629..6653
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7118..7768
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7956..7973)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8207..8226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8449..8558
                     /note="incP origin of transfer"
     CDS             8591..8962
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      9089..9804
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10427..11194
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11386..12180
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12479..13627
                     /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    15223..15247
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15494..15515)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15706..15725
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15865..15881
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15894..16146)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16290..16841)
                     /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(16862..17207)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16899..16919)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17743..17759
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aatctgagtt tttctgatta acagggtacc gggccccccc tcgaggcgcg ccaagctatc
       61 aaacaagttt gtacaaaaaa gcaggctccg aattcgccct tcaccatgga ttacaaggat
      121 gatgatgata aggattacaa ggatgatgat gataagatgg ctccaaagaa gaagagaaag
      181 gttggaatcc acggagttcc agctgctgat aagaagtact ctatcggact tgacatcgga
      241 accaactctg ttggatgggc tgttatcacc gatgagtaca aggttccatc taagaagttc
      301 aaggttcttg gaaacaccga tagacactct atcaagaaga accttatcgg tgctcttctt
      361 ttcgattctg gagagaccgc tgaggctacc agattgaaga gaaccgctag aagaagatac
      421 accagaagaa agaacagaat ctgctacctt caggaaatct tctctaacga gatggctaag
      481 gttgatgatt ctttcttcca cagacttgag gagtctttcc ttgttgagga ggataagaag
      541 cacgagagac acccaatctt cggaaacatc gttgatgagg ttgcttacca cgagaagtac
      601 ccaaccatct accaccttag aaagaagttg gttgattcta ccgataaggc tgatcttaga
      661 cttatctacc ttgctcttgc tcacatgatc aagttcagag gacacttcct tatcgaggga
      721 gaccttaacc cagataactc tgatgttgat aagttgttca tccagcttgt tcagacctac
      781 aaccagcttt tcgaggagaa cccaatcaac gcttctggag ttgatgctaa ggctatcctt
      841 tctgctagac tttctaagtc tcgtagactt gagaacctta tcgctcagct tccaggagag
      901 aagaagaacg gacttttcgg aaaccttatc gctctttctc ttggacttac cccaaacttc
      961 aagtctaact tcgatcttgc tgaggatgct aagttgcagc tttctaagga tacctacgat
     1021 gatgatcttg ataaccttct tgctcagatc ggagatcagt acgctgatct tttccttgct
     1081 gctaagaacc tttctgatgc tatccttctt tctgacatcc ttagagttaa caccgagatc
     1141 accaaggctc cactttctgc ttctatgatc aagagatacg atgagcacca ccaggatctt
     1201 acccttttga aggctcttgt tagacagcag cttccagaga agtacaagga aatcttcttc
     1261 gatcagtcta agaacggata cgctggatac atcgatggag gagcttctca ggaggagttc
     1321 tacaagttca tcaagccaat ccttgagaag atggatggaa ccgaggagct tcttgttaag
     1381 ttgaacagag aggatcttct tagaaagcag agaaccttcg ataacggatc tatcccacac
     1441 cagatccacc ttggagagct tcacgctatc cttcgtagac aggaggattt ctacccattc
     1501 ttgaaggata acagagagaa gatcgagaag atccttacct tcagaatccc atactacgtt
     1561 ggaccacttg ctagaggaaa ctctcgtttc gcttggatga ccagaaagtc tgaggagacc
     1621 atcacccctt ggaacttcga ggaggtaagt ttctgcttct acctttgata tatatataat
     1681 aattatcatt aattagtagt aatataatat ttcaaatatt tttttcaaaa taaaagaatg
     1741 tagtatatag caattgcttt tctgtagttt ataagtgtgt atattttaat ttataacttt
     1801 tctaatatat gaccaaaatt tgttgatgtg caggttgttg ataagggagc ttctgctcag
     1861 tctttcatcg agagaatgac caacttcgat aagaaccttc caaacgagaa ggttcttcca
     1921 aagcactctc ttctttacga gtacttcacc gtttacaacg agcttaccaa ggttaagtac
     1981 gttaccgagg gaatgagaaa gccagctttc ctttctggag agcagaagaa ggctatcgtt
     2041 gatcttcttt tcaagaccaa cagaaaggtt accgttaagc agttgaagga ggattacttc
     2101 aagaagatcg agtgcttcga ttctgttgaa atctctggag ttgaggatag attcaacgct
     2161 tctcttggaa cctaccacga tcttttgaag atcatcaagg ataaggattt ccttgataac
     2221 gaggagaacg aggacatcct tgaggacatc gttcttaccc ttaccctttt cgaggataga
     2281 gagatgatcg aggagagact caagacctac gctcaccttt tcgatgataa ggttatgaag
     2341 cagttgaaga gaagaagata caccggatgg ggtagacttt ctcgtaagtt gatcaacgga
     2401 atcagagata agcagtctgg aaagaccatc cttgatttct tgaagtctga tggattcgct
     2461 aacagaaact tcatgcagct tatccacgat gattctctta ccttcaagga ggacatccag
     2521 aaggctcagg tttctggaca gggagattct cttcacgagc acatcgctaa ccttgctgga
     2581 tctccagcta tcaagaaggg aatccttcag accgttaagg ttgttgatga gcttgttaag
     2641 gttatgggta gacacaagcc agagaacatc gttatcgaga tggctagaga gaaccagacc
     2701 acccagaagg gacagaagaa ctctcgtgag agaatgaaga gaatcgagga gggaatcaag
     2761 gagcttggat ctcaaatctt gaaggagcac ccagttgaga acacccagct tcagaacgag
     2821 aagttgtacc tttactacct tcagaacgga agagatatgt acgttgatca ggagcttgac
     2881 atcaacagac tttctgatta cgatgttgat cacatcgttc cacagtcttt cttgaaggat
     2941 gattctatcg ataacaaggt tcttacccgt tctgataaga acagaggaaa gtctgataac
     3001 gttccatctg aggaggttgt taagaagatg aagaactact ggagacagct tcttaacgct
     3061 aagttgatca cccagagaaa gttcgataac cttaccaagg ctgagagagg aggactttct
     3121 gagcttgata aggctggatt catcaagaga cagcttgttg agaccagaca gatcaccaag
     3181 cacgttgctc agatccttga ttctcgtatg aacaccaagt acgatgagaa cgataagttg
     3241 atcagagagg ttaaggttat caccttgaag tctaagttgg tttctgattt cagaaaggat
     3301 ttccagttct acaaggttag agagatcaac aactaccacc acgctcacga tgcttacctt
     3361 aacgctgttg ttggaaccgc tcttatcaag aagtacccaa agttggagtc tgagttcgtt
     3421 tacggagatt acaaggttta cgatgttaga aagatgatcg ctaagtctga gcaggagatc
     3481 ggaaaggcta ccgctaagta cttcttctac tctaacatca tgaacttctt caagaccgag
     3541 atcacccttg ctaacggaga gatcagaaag agaccactta tcgagaccaa cggagagacc
     3601 ggagagatcg tttgggataa gggaagagat ttcgctaccg ttagaaaggt tctttctatg
     3661 ccacaggtta acatcgttaa gaaaaccgag gttcagaccg gaggattctc taaggagtct
     3721 atccttccaa agagaaactc tgataagttg atcgctagaa agaaggattg ggacccaaag
     3781 aagtacggag gattcgattc tccaaccgtt gcttactctg ttcttgttgt tgctaaggtt
     3841 gagaagggaa agtctaagaa gttgaagtct gttaaggagc ttcttggaat caccatcatg
     3901 gagcgttctt ctttcgagaa gaacccaatc gatttccttg aggctaaggg atacaaggag
     3961 gttaagaagg atcttatcat caagttgcca aagtactctc ttttcgagct tgagaacgga
     4021 agaaagagaa tgcttgcttc tgctggagag cttcagaagg gaaacgagct tgctcttcca
     4081 tctaagtacg ttaacttcct ttaccttgct tctcactacg agaagttgaa gggatctcca
     4141 gaggataacg agcagaagca gcttttcgtt gagcagcaca agcactacct tgatgagatc
     4201 atcgagcaaa tctctgagtt ctctaagaga gttatccttg ctgatgctaa ccttgataag
     4261 gttctttctg cttacaacaa gcacagagat aagccaatca gagagcaggc tgagaacatc
     4321 atccaccttt tcacccttac caaccttggt gctccagctg ctttcaagta cttcgatacc
     4381 accatcgata gaaaaagata cacctctacc aaggaggttc ttgatgctac ccttatccac
     4441 cagtctatca ccggacttta cgagaccaga atcgatcttt ctcagcttgg aggagataag
     4501 agaccagctg ctaccaagaa ggctggacag gctaagaaga agaagtgaga cgtccgatcg
     4561 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
     4621 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
     4681 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
     4741 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
     4801 actagatcgg gaattgatcc cccctcgaca gcttccggaa agggcgaatt cgcaactttg
     4861 tatacaaaag ttgccgagct cgcggccgca tgttgttgtt accagaaagt aaataaatgt
     4921 tcaatctctg atgttctcaa gtaagtgagt tttattggga ataatattaa cttatgttct
     4981 tcttgcattt gatttctttg ccgctctctt cttctatctt aaatctgtgt atactatttc
     5041 actattgggc tttttattag tctataatgg gactcaaaat aaggctttgg cccacatcaa
     5101 aaagataagt cacaaatcaa aactaaattc agagtctttt ctcccacatc ggtcactgta
     5161 ctcattttgt gtttgtttat atattacacg aaccgatctt tggtacgttc caccatcacc
     5221 acaaccagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     5281 aaaaagtggc accgagtcgg tgcttttttt ctagacccag ctttcttgta caaagttggc
     5341 attaccatgg cttcgttgaa caacggaaac tcgacttgcc ttccgcacaa tacatcattt
     5401 cttcttagct ttttttcttc ttcttcgttc atacagtttt tttttgttta tcagcttaca
     5461 ttttcttgaa ccgtagcttt cgttttcttc tttttaactt tccattcgga gtttttgtat
     5521 cttgtttcat agtttgtccc aggattagaa tgattaggca tcgaaccttc aagaatttga
     5581 ttgaataaaa catcttcatt cttaagatat gaagataatc ttcaaaaggc ccctgggaat
     5641 ctgaaagaag agaagcaggc ccatttatat gggaaagaac aatagtattt cttatatagg
     5701 cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg cttagataag aaaacgaagc
     5761 tgagtttata tacagctaga gtcgaagtag tgattggatt tgtactacga gtttggtttt
     5821 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     5881 cgagtcggtg ctttttttct agacccagct ttcttgtaca aagttggcat tactgcaggt
     5941 cgacgaccca gctttcttgt acaaagttgg cattactgca ggtcgacgac ccagctttct
     6001 tgtacaaagt ggttcgataa ttcttaatta actagttcta gagcggccgc caccgcggtg
     6061 gagctcgaat ttccccgatc gttcaaacat ttggcaataa agtttcttaa gattgaatcc
     6121 tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta agcatgtaat
     6181 aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta gagtcccgca
     6241 attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg ataaattatc
     6301 gcgcgcggtg tcatctatgt tactagatcg ggaattcgta atcatggtca tagctgtttc
     6361 ctgcctagtc ggagcggccg cgtccgtcta gctgatagtg accttaggcg acttttgaac
     6421 gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa cccgcggctg
     6481 agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct tgtcccgcgt
     6541 catcggcggg ggtcataacg tgactccctt aattctccgc tcatgatcag attgtcgttt
     6601 cccgccttca gtttaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag
     6661 aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc
     6721 gtccatttgt atgtgcatgc caaccacagg gttccccaga tctggcgccg gccagcgaga
     6781 cgagcaagat tggccgccgc ccgaaacgat ccgacagcgc gcccagcaca ggtgcgcagg
     6841 caaattgcac caacgcatac agcgccagca gaatgccata gtgggcggtg acgtcgttcg
     6901 agtgaaccag atcgcgcagg aggcccggca gcaccggcat aatcaggccg atgccgacag
     6961 cgtcgagcgc gacagtgctc agaattacga tcaggggtat gttgggtttc acgtctggcc
     7021 tccggaccag cctccgctgg tccgattgaa cgcgcggatt ctttatcact gataagttgg
     7081 tggacatatt atgtttatca gtgataaagt gtcaagcatg acaaagttgc agccgaatac
     7141 agtgatccgt gccgccctgg acctgttgaa cgaggtcggc gtagacggtc tgacgacacg
     7201 caaactggcg gaacggttgg gggttcagca gccggcgctt tactggcact tcaggaacaa
     7261 gcgggcgctg ctcgacgcac tggccgaagc catgctggcg gagaatcata cgcattcggt
     7321 gccgagagcc gacgacgact ggcgctcatt tctgatcggg aatgcccgca gcttcaggca
     7381 ggcgctgctc gcctaccgcg atggcgcgcg catccatgcc ggcacgcgac cgggcgcacc
     7441 gcagatggaa acggccgacg cgcagcttcg cttcctctgc gaggcgggtt tttcggccgg
     7501 ggacgccgtc aatgcgctga tgacaatcag ctacttcact gttggggccg tgcttgagga
     7561 gcaggccggc gacagcgatg ccggcgagcg cggcggcacc gttgaacagg ctccgctctc
     7621 gccgctgttg cgggccgcga tagacgcctt cgacgaagcc ggtccggacg cagcgttcga
     7681 gcagggactc gcggtgattg tcgatggatt ggcgaaaagg aggctcgttg tcaggaacgt
     7741 tgaaggaccg agaaagggtg acgattgatc aggaccgctg ccggagcgca acccactcac
     7801 tacagcagag ccatgtagac aacatcccct ccccctttcc accgcgtcag acgcccgtag
     7861 cagcccgcta cgggcttttt catgccctgc cctagcgtcc aagcctcacg gccgcgctcg
     7921 gcctctctgg cggccttctg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7981 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     8041 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     8101 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     8161 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     8221 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8281 ctgtccgcct ttctcccttc gggaagcgtg gcgcttttcc gctgcataac cctgcttcgg
     8341 ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg
     8401 ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg
     8461 tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc
     8521 ggtgctcaac gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga
     8581 gggcaagcgg atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt
     8641 gtactgcctt ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag
     8701 cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa atcacgggcg tcgtggacta
     8761 tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct
     8821 gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc
     8881 cctgctggcg aagatcgaag agaagcagga cgagcttggc aaggtcatga tgggcgtggt
     8941 ccgcccgagg gcagagccat gactttttta gccgctaaaa cggccggggg gtgcgcgtga
     9001 ttgccaagca cgtccccatg cgctccatca agaagagcga cttcgcggag ctggtgaagt
     9061 acatcaccga cgagcaaggc aagaccgagc gcctttccga cgctcaccgg gctggttgcc
     9121 ctcgccgctg ggctggcggc cgtctatggc cctgcaaacg cgccagaaac gccgtcgaag
     9181 ccgtgtgcga gacaccgcgg ccggccgccg gcgttgtgga tacctcgcgg aaaacttggc
     9241 cctcactgac agatgagggg cggacgttga cacttgaggg gccgactcac ccggcgcggc
     9301 gttgacagat gaggggcagg ctcgatttcg gccggcgacg tggagctggc cagcctcgca
     9361 aatcggcgaa aacgcctgat tttacgcgag tttcccacag atgatgtgga caagcctggg
     9421 gataagtgcc ctgcggtatt gacacttgag gggcgcgact actgacagat gaggggcgcg
     9481 atccttgaca cttgaggggc agagtgctga cagatgaggg gcgcacctat tgacatttga
     9541 ggggctgtcc acaggcagaa aatccagcat ttgcaagggt ttccgcccgt ttttcggcca
     9601 ccgctaacct gtcttttaac ctgcttttaa accaatattt ataaaccttg tttttaacca
     9661 gggctgcgcc ctgtgcgcgt gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac
     9721 cctcccggcc cgctaacgcg ggcctcccat ccccccaggg gctgcgcccc tcggccgcga
     9781 acggcctcac cccaaaaatg gcagcgctgg cagtccttgc cattgccggg atcggggcag
     9841 taacgggatg ggcgatcagc ccgagcgcga cgcccggaag cattgacgtg ccgcaggtgc
     9901 tggcatcgac attcagcgac caggtgccgg gcagtgaggg cggcggcctg ggtggcggcc
     9961 tgcccttcac ttcggccgtc ggggcattca cggacttcat ggcggggccg gcaattttta
    10021 ccttgggcat tcttggcata gtggtcgcgg gtgccgtgct cgtgttcggg ggtgcgataa
    10081 acccagcgaa ccatttgagg tgataggtaa gattataccg aggtatgaaa acgagaattg
    10141 gacctttaca gaattactct atgaagcgcc atatttaaaa agctaccaag acgaagagga
    10201 tgaagaggat gaggaggcag attgccttga atatattgac aatactgata agataatata
    10261 tcttttatat agaagatatc gccgtatgta aggatttcag ggggcaaggc ataggcagcg
    10321 cgcttatcaa tatatctata gaatgggcaa agcataaaaa cttgcatgga ctaatgcttg
    10381 aaacccagga caataacctt atagcttgta aattctatca taattgggta atgactccaa
    10441 cttattgata gtgttttatg ttcagataat gcccgatgac tttgtcatgc agctccaccg
    10501 attttgagaa cgacagcgac ttccgtccca gccgtgccag gtgctgcctc agattcaggt
    10561 tatgccgctc aattcgctgc gtatatcgct tgctgattac gtgcagcttt cccttcaggc
    10621 gggattcata cagcggccag ccatccgtca tccatatcac cacgtcaaag ggtgacagca
    10681 ggctcataag acgccccagc gtcgccatag tgcgttcacc gaatacgtgc gcaacaaccg
    10741 tcttccggag actgtcatac gcgtaaaaca gccagcgctg gcgcgattta gccccgacat
    10801 agccccactg ttcgtccatt tccgcgcaga cgatgacgtc actgcccggc tgtatgcgcg
    10861 aggttaccga ctgcggcctg agttttttaa gtgacgtaaa atcgtgttga ggccaacgcc
    10921 cataatgcgg gctgttgccc ggcatccaac gccattcatg gccatatcaa tgattttctg
    10981 gtgcgtaccg ggttgagaag cggtgtaagt gaactgcagt tgccatgttt tacggcagtg
    11041 agagcagaga tagcgctgat gtccggcggt gcttttgccg ttacgcacca ccccgtcagt
    11101 agctgaacag gagggacagc tgatagaaac agaagccact ggagcacctc aaaaacacca
    11161 tcatacacta aatcagtaag ttggcagcat cacccataat tgtggtttca aaatcggctc
    11221 cgtcgatact atgttatacg ccaactttga aaacaacttt gaaaaagctg ttttctggta
    11281 tttaaggttt tagaatgcaa ggaacagtga attggagttc gtcttgttat aattagcttc
    11341 ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc taaaatgaga
    11401 atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga tacggaagga
    11461 atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata tttaaaaatg
    11521 acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga catgatgcta
    11581 tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca tgatggctgg
    11641 agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta tgaagatgaa
    11701 caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt tcactccatc
    11761 gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga attggattac
    11821 ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga cactccattt
    11881 aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga ggaacttgtc
    11941 ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa agtaagtggc
    12001 tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc cttctgcgtc
    12061 cggtcgatca gggaggatat cggggaagaa cagtatgtcg agctattttt tgacttactg
    12121 gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga attgttttag
    12181 tacctagatg tggcgcaacg atgccggcga caagcaggag cgcaccgact tcttccgcat
    12241 caagtgtttt ggctctcagg ccgaggccca cggcaagtat ttgggcaagg ggtcgctggt
    12301 attcgtgcag ggcaagattc ggaataccaa gtacgagaag gacggccaga cggtctacgg
    12361 gaccgacttc attgccgata aggtggatta tctggacacc aaggcaccag gcgggtcaaa
    12421 tcaggaataa gggcacattg ccccggcgtg agtcggggca atcccgcaag gagggtgaat
    12481 gaatcggacg tttgaccgga aggcatacag gcaagaactg atcgacgcgg ggttttccgc
    12541 cgaggatgcc gaaaccatcg caagccgcac cgtcatgcgt gcgccccgcg aaaccttcca
    12601 gtccgtcggc tcgatggtcc agcaagctac ggccaagatc gagcgcgaca gcgtgcaact
    12661 ggctccccct gccctgcccg cgccatcggc cgccgtggag cgttcgcgtc gtctcgaaca
    12721 ggaggcggca ggtttggcga agtcgatgac catcgacacg cgaggaacta tgacgaccaa
    12781 gaagcgaaaa accgccggcg aggacctggc aaaacaggtc agcgaggcca agcaggccgc
    12841 gttgctgaaa cacacgaagc agcagatcaa ggaaatgcag ctttccttgt tcgatattgc
    12901 gccgtggccg gacacgatgc gagcgatgcc aaacgacacg gcccgctctg ccctgttcac
    12961 cacgcgcaac aagaaaatcc cgcgcgaggc gctgcaaaac aaggtcattt tccacgtcaa
    13021 caaggacgtg aagatcacct acaccggcgt cgagctgcgg gccgacgatg acgaactggt
    13081 gtggcagcag gtgttggagt acgcgaagcg cacccctatc ggcgagccga tcaccttcac
    13141 gttctacgag ctttgccagg acctgggctg gtcgatcaat ggccggtatt acacgaaggc
    13201 cgaggaatgc ctgtcgcgcc tacaggcgac ggcgatgggc ttcacgtccg accgcgttgg
    13261 gcacctggaa tcggtgtcgc tgctgcaccg cttccgcgtc ctggaccgtg gcaagaaaac
    13321 gtcccgttgc caggtcctga tcgacgagga aatcgtcgtg ctgtttgctg gcgaccacta
    13381 cacgaaattc atatgggaga agtaccgcaa gctgtcgccg acggcccgac ggatgttcga
    13441 ctatttcagc tcgcaccggg agccgtaccc gctcaagctg gaaaccttcc gcctcatgtg
    13501 cggatcggat tccacccgcg tgaagaagtg gcgcgagcag gtcggcgaag cctgcgaaga
    13561 gttgcgaggc agcggcctgg tggaacacgc ctgggtcaat gatgacctgg tgcattgcaa
    13621 acgctagggc cttgtggggt cagttccggc tgggggttca gcagccagcg ctttactggc
    13681 atttcaggaa caagcgggca ctgctcgacg cacttgcttc gctcagtatc gctcgggacg
    13741 cacggcgcgc tctacgaact gccgataaac agaggattaa aattgacaat tgtgattaag
    13801 gctcagattc gacggcttgg agcggccgac gtgcaggatt tccgcgagat ccgattgtcg
    13861 gccctgaaga aagctccaga gatgttcggg tccgtttacg agcacgagga gaaaaagccc
    13921 atggaggcgt tcgctgaacg gttgcgagat gccgtggcat tcggcgccta catcgacggc
    13981 gagatcattg ggctgtcggt cttcaaacag gaggacggcc ccaaggacgc tcacaaggcg
    14041 catctgtccg gcgttttcgt ggagcccgaa cagcgaggcc gaggggtcgc cggtatgctg
    14101 ctgcgggcgt tgccggcggg tttattgctc gtgatgatcg tccgacagat tccaacggga
    14161 atctggtgga tgcgcatctt catcctcggc gcacttaata tttcgctatt ctggagcttg
    14221 ttgtttattt cggtctaccg cctgccgggc ggggtcgcgg cgacggtagg cgctgtgcag
    14281 ccgctgatgg tcgtgttcat ctctgccgct ctgctaggta gcccgatacg attgatggcg
    14341 gtcctggggg ctatttgcgg aactgcgggc gtggcgctgt tggtgttgac accaaacgca
    14401 gcgctagatc ctgtcggcgt cgcagcgggc ctggcggggg cggtttccat ggcgttcgga
    14461 accgtgctga cccgcaagtg gcaacctccc gtgcctctgc tcacctttac cgcctggcaa
    14521 ctggcggccg gaggacttct gctcgttcca gtagctttag tgtttgatcc gccaatcccg
    14581 atgcctacag gaaccaatgt tctcggcctg gcgtggctcg gcctgatcgg agcgggttta
    14641 acctacttcc tttggttccg ggggatctcg cgactcgaac ctacagttgt ttccttactg
    14701 ggctttctca gccccagatc tggggtcgat cagccgggga tgcatcaggc cgacagtcgg
    14761 aacttcgggt ccccgacctg taccattcgg tgagcaatgg ataggggagt tgatatcgtc
    14821 aacgttcact tctaaagaaa tagcgccact cagcttcctc agcggcttta tccagcgatt
    14881 tcctattatg tcggcatagt tctcaagatc gacagcctgt cacggttaag cgagaaatga
    14941 ataagaaggc tgataattcg gatctctgcg agggagatga tatttgatca caggcagcaa
    15001 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc
    15061 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct
    15121 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga
    15181 ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg
    15241 tgtaaacaaa ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg
    15301 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
    15361 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
    15421 tgatggtggt tccgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagatagg
    15481 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
    15541 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
    15601 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
    15661 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
    15721 aggagcgggc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
    15781 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
    15841 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattaatt cccgatctag
    15901 taacatagat gacaccgcgc gcgataattt atcctagttt gcgcgctata ttttgttttc
    15961 tatcgcgtat taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa
    16021 cgtcatgcat tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat
    16081 aatcatcgca agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga
    16141 acgatcgggg aaattcgagc tcggtacccc ctcctgggca cggtcgtacg cgatccgccg
    16201 cggttccacc tcaccattgc gggtatgcca gttgagttcg atgagttccc gtaccagctc
    16261 caactcgggc gatgccacgc gcactccgtt cagatctcgg tgacgggcag gaccggacgg
    16321 ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca cgtcatgcca gttcccgtgc
    16381 ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc cgagcgcctc gtgcatgcgc
    16441 acgctcgggt cgttgggcag cccgatgaca gcgaccacgc tcttgaagcc ctgtgcctcc
    16501 agggacttca gcaggtgggt gtagagcgtg gagcccagtc ccgtccgctg gtggcggggg
    16561 gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt tgcgtgcctt ccaggggccc
    16621 gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga cgagccaggg atagcgctcc
    16681 cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct gcggctcggt acggaagttg
    16741 accgtgcttg tctcgatgta gtggttgacg atggtgcaga ccgccggcat gtccgcctcg
    16801 gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca tggatcctct agagtccccc
    16861 gtgttctctc caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt
    16921 gggattgtgc gtcatccctt acgtcagtgg agatatcaca tcaatccact tgctttgaag
    16981 acgtggttgg aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg
    17041 gaccactgtc ggcagaggca tcttcaacga tggcctttcc tttatcgcaa tgatggcatt
    17101 tgtaggagcc accttccttt tccactatct tcacaataaa gtgacagata gctgggcaat
    17161 ggaatccgag gaggtttccg gatattaccc tttgttgaaa agtctcaatt gccctttggt
    17221 cttctgagac tgtatctttg atatttttgg agtagacaag tgtgtcgtgc tccaccatgt
    17281 tgacgaagat tttcttcttg tcattgagtc gtaagagact ctgtatgaac tgttcgccag
    17341 tctttacggc gagttctgtt aggtcctcta tttgaatctt tgactccatg gcctttgatt
    17401 cagtgggaac taccttttta gagactccaa tctctattac ttgccttggt ttgtgaagca
    17461 agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatata tctttctctg
    17521 tgttcttgat gcagttagtc ctgaatcttt tgactgcatc tttaaccttc ttgggaaggt
    17581 atttgatttc ctggagatta ttgctcgggt agatcgtctt gatgagacct gctgcgtaag
    17641 cctctctaac catctgtggg ttagcattct ttctgaaatt gaaaaggcta atctggggac
    17701 ctgcaggggc aataccgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
    17761 ccaagctttc gacgagtcag taataaacgg cgtcaaagtg gttgcagccg gcacacacga
    17821 gtcgtgttta tcaactcaaa gcacaaatac ttttcctcaa cctaaaaata aggcaattag
    17881 ccaaaaacaa ctttgcgtgt aaacaacgct caatacacgt gtcattttat tattagctat
    17941 tgcttcaccg ccttagcttt ctcgtgacct agtcgtcctc gtcttttctt cttcttcttc
    18001 tataaaacaa tacccaaaga gctcttcttc ttcacaattc agatttcaat ttctcaaaat
    18061 cttaaaaact ttctctcaat tctctctacc gtgatcaagg taaatttctg tgttccttat
    18121 tctctcaaaa tcttcgattt tgttttcgtt cgatcccaat ttcgtatatg ttctttggtt
    18181 tagattctgt taatcttaga tcgaagacga ttttctgggt ttgatcgtta gatatcatct
    18241 taattctcga ttagggtttc atagatatca tccgatttgt tcaaataatt tgagttttgt
    18301 cgaataatta ctcttcgatt tgtgatttct atctagatct ggtgttagtt tctagtttgt
    18361 gcgatcgaat ttgtagatt
//
