LOCUS       sequence-202699-       18336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101120, Medtr2g101130 
            (Acre2). Also expresses Cas9 from rolD promoter from AtUBQ10 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18336)
  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 18336)
  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: pSC24
FEATURES             Location/Qualifiers
     source          1..18336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    75..99
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             121..168
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             175..195
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(220..1656,1846..4509)
                     /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          1657..1845
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4510..4557
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4568..4820
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4865..4885
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5240..5315
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5327..5351)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5828..5903
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5915..5939)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5958..5982)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6046..6298
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6316..6332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6316..6332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6598..6622
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7087..7737
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7925..7942)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8176..8195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8418..8527
                     /note="incP origin of transfer"
     CDS             8560..8931
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      9058..9773
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10396..11163
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11355..12149
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12448..13596
                     /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    15192..15216
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15463..15484)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15675..15694
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15834..15850
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15863..16115)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16259..16810)
                     /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(16831..17176)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16868..16888)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17712..17728
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aatttgtaga ttaatctgag tttttctgat taacagggta ccgggccccc cctcgaggcg
       61 cgccaagcta tcaaacaagt ttgtacaaaa aagcaggctc cgaattcgcc cttcaccatg
      121 gattacaagg atgatgatga taaggattac aaggatgatg atgataagat ggctccaaag
      181 aagaagagaa aggttggaat ccacggagtt ccagctgctg ataagaagta ctctatcgga
      241 cttgacatcg gaaccaactc tgttggatgg gctgttatca ccgatgagta caaggttcca
      301 tctaagaagt tcaaggttct tggaaacacc gatagacact ctatcaagaa gaaccttatc
      361 ggtgctcttc ttttcgattc tggagagacc gctgaggcta ccagattgaa gagaaccgct
      421 agaagaagat acaccagaag aaagaacaga atctgctacc ttcaggaaat cttctctaac
      481 gagatggcta aggttgatga ttctttcttc cacagacttg aggagtcttt ccttgttgag
      541 gaggataaga agcacgagag acacccaatc ttcggaaaca tcgttgatga ggttgcttac
      601 cacgagaagt acccaaccat ctaccacctt agaaagaagt tggttgattc taccgataag
      661 gctgatctta gacttatcta ccttgctctt gctcacatga tcaagttcag aggacacttc
      721 cttatcgagg gagaccttaa cccagataac tctgatgttg ataagttgtt catccagctt
      781 gttcagacct acaaccagct tttcgaggag aacccaatca acgcttctgg agttgatgct
      841 aaggctatcc tttctgctag actttctaag tctcgtagac ttgagaacct tatcgctcag
      901 cttccaggag agaagaagaa cggacttttc ggaaacctta tcgctctttc tcttggactt
      961 accccaaact tcaagtctaa cttcgatctt gctgaggatg ctaagttgca gctttctaag
     1021 gatacctacg atgatgatct tgataacctt cttgctcaga tcggagatca gtacgctgat
     1081 cttttccttg ctgctaagaa cctttctgat gctatccttc tttctgacat ccttagagtt
     1141 aacaccgaga tcaccaaggc tccactttct gcttctatga tcaagagata cgatgagcac
     1201 caccaggatc ttaccctttt gaaggctctt gttagacagc agcttccaga gaagtacaag
     1261 gaaatcttct tcgatcagtc taagaacgga tacgctggat acatcgatgg aggagcttct
     1321 caggaggagt tctacaagtt catcaagcca atccttgaga agatggatgg aaccgaggag
     1381 cttcttgtta agttgaacag agaggatctt cttagaaagc agagaacctt cgataacgga
     1441 tctatcccac accagatcca ccttggagag cttcacgcta tccttcgtag acaggaggat
     1501 ttctacccat tcttgaagga taacagagag aagatcgaga agatccttac cttcagaatc
     1561 ccatactacg ttggaccact tgctagagga aactctcgtt tcgcttggat gaccagaaag
     1621 tctgaggaga ccatcacccc ttggaacttc gaggaggtaa gtttctgctt ctacctttga
     1681 tatatatata ataattatca ttaattagta gtaatataat atttcaaata tttttttcaa
     1741 aataaaagaa tgtagtatat agcaattgct tttctgtagt ttataagtgt gtatatttta
     1801 atttataact tttctaatat atgaccaaaa tttgttgatg tgcaggttgt tgataaggga
     1861 gcttctgctc agtctttcat cgagagaatg accaacttcg ataagaacct tccaaacgag
     1921 aaggttcttc caaagcactc tcttctttac gagtacttca ccgtttacaa cgagcttacc
     1981 aaggttaagt acgttaccga gggaatgaga aagccagctt tcctttctgg agagcagaag
     2041 aaggctatcg ttgatcttct tttcaagacc aacagaaagg ttaccgttaa gcagttgaag
     2101 gaggattact tcaagaagat cgagtgcttc gattctgttg aaatctctgg agttgaggat
     2161 agattcaacg cttctcttgg aacctaccac gatcttttga agatcatcaa ggataaggat
     2221 ttccttgata acgaggagaa cgaggacatc cttgaggaca tcgttcttac ccttaccctt
     2281 ttcgaggata gagagatgat cgaggagaga ctcaagacct acgctcacct tttcgatgat
     2341 aaggttatga agcagttgaa gagaagaaga tacaccggat ggggtagact ttctcgtaag
     2401 ttgatcaacg gaatcagaga taagcagtct ggaaagacca tccttgattt cttgaagtct
     2461 gatggattcg ctaacagaaa cttcatgcag cttatccacg atgattctct taccttcaag
     2521 gaggacatcc agaaggctca ggtttctgga cagggagatt ctcttcacga gcacatcgct
     2581 aaccttgctg gatctccagc tatcaagaag ggaatccttc agaccgttaa ggttgttgat
     2641 gagcttgtta aggttatggg tagacacaag ccagagaaca tcgttatcga gatggctaga
     2701 gagaaccaga ccacccagaa gggacagaag aactctcgtg agagaatgaa gagaatcgag
     2761 gagggaatca aggagcttgg atctcaaatc ttgaaggagc acccagttga gaacacccag
     2821 cttcagaacg agaagttgta cctttactac cttcagaacg gaagagatat gtacgttgat
     2881 caggagcttg acatcaacag actttctgat tacgatgttg atcacatcgt tccacagtct
     2941 ttcttgaagg atgattctat cgataacaag gttcttaccc gttctgataa gaacagagga
     3001 aagtctgata acgttccatc tgaggaggtt gttaagaaga tgaagaacta ctggagacag
     3061 cttcttaacg ctaagttgat cacccagaga aagttcgata accttaccaa ggctgagaga
     3121 ggaggacttt ctgagcttga taaggctgga ttcatcaaga gacagcttgt tgagaccaga
     3181 cagatcacca agcacgttgc tcagatcctt gattctcgta tgaacaccaa gtacgatgag
     3241 aacgataagt tgatcagaga ggttaaggtt atcaccttga agtctaagtt ggtttctgat
     3301 ttcagaaagg atttccagtt ctacaaggtt agagagatca acaactacca ccacgctcac
     3361 gatgcttacc ttaacgctgt tgttggaacc gctcttatca agaagtaccc aaagttggag
     3421 tctgagttcg tttacggaga ttacaaggtt tacgatgtta gaaagatgat cgctaagtct
     3481 gagcaggaga tcggaaaggc taccgctaag tacttcttct actctaacat catgaacttc
     3541 ttcaagaccg agatcaccct tgctaacgga gagatcagaa agagaccact tatcgagacc
     3601 aacggagaga ccggagagat cgtttgggat aagggaagag atttcgctac cgttagaaag
     3661 gttctttcta tgccacaggt taacatcgtt aagaaaaccg aggttcagac cggaggattc
     3721 tctaaggagt ctatccttcc aaagagaaac tctgataagt tgatcgctag aaagaaggat
     3781 tgggacccaa agaagtacgg aggattcgat tctccaaccg ttgcttactc tgttcttgtt
     3841 gttgctaagg ttgagaaggg aaagtctaag aagttgaagt ctgttaagga gcttcttgga
     3901 atcaccatca tggagcgttc ttctttcgag aagaacccaa tcgatttcct tgaggctaag
     3961 ggatacaagg aggttaagaa ggatcttatc atcaagttgc caaagtactc tcttttcgag
     4021 cttgagaacg gaagaaagag aatgcttgct tctgctggag agcttcagaa gggaaacgag
     4081 cttgctcttc catctaagta cgttaacttc ctttaccttg cttctcacta cgagaagttg
     4141 aagggatctc cagaggataa cgagcagaag cagcttttcg ttgagcagca caagcactac
     4201 cttgatgaga tcatcgagca aatctctgag ttctctaaga gagttatcct tgctgatgct
     4261 aaccttgata aggttctttc tgcttacaac aagcacagag ataagccaat cagagagcag
     4321 gctgagaaca tcatccacct tttcaccctt accaaccttg gtgctccagc tgctttcaag
     4381 tacttcgata ccaccatcga tagaaaaaga tacacctcta ccaaggaggt tcttgatgct
     4441 acccttatcc accagtctat caccggactt tacgagacca gaatcgatct ttctcagctt
     4501 ggaggagata agagaccagc tgctaccaag aaggctggac aggctaagaa gaagaagtga
     4561 gacgtccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     4621 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     4681 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
     4741 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
     4801 gtcatctatg ttactagatc gggaattgat cccccctcga cagcttccgg aaagggcgaa
     4861 ttcgcaactt tgtatacaaa agttgccgag ctcgcggccg catgttgttg ttaccagaaa
     4921 gtaaataaat gttcaatctc tgatgttctc aagtaagtga gttttattgg gaataatatt
     4981 aacttatgtt cttcttgcat ttgatttctt tgccgctctc ttcttctatc ttaaatctgt
     5041 gtatactatt tcactattgg gctttttatt agtctataat gggactcaaa ataaggcttt
     5101 ggcccacatc aaaaagataa gtcacaaatc aaaactaaat tcagagtctt ttctcccaca
     5161 tcggtcactg tactcatttt gtgtttgttt atatattaca cgaaccgatc tttggtacgc
     5221 ccaatggtga ggcaattgag ttttagagct agaaatagca agttaaaata aggctagtcc
     5281 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg
     5341 tacaaagttg gcattaccat ggcttcgttg aacaacggaa actcgacttg ccttccgcac
     5401 aatacatcat ttcttcttag ctttttttct tcttcttcgt tcatacagtt tttttttgtt
     5461 tatcagctta cattttcttg aaccgtagct ttcgttttct tctttttaac tttccattcg
     5521 gagtttttgt atcttgtttc atagtttgtc ccaggattag aatgattagg catcgaacct
     5581 tcaagaattt gattgaataa aacatcttca ttcttaagat atgaagataa tcttcaaaag
     5641 gcccctggga atctgaaaga agagaagcag gcccatttat atgggaaaga acaatagtat
     5701 ttcttatata ggcccattta agttgaaaac aatcttcaaa agtcccacat cgcttagata
     5761 agaaaacgaa gctgagttta tatacagcta gagtcgaagt agtgattgcc accaatttac
     5821 cgcatcagtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     5881 aaaaagtggc accgagtcgg tgcttttttt ctagacccag ctttcttgta caaagttggc
     5941 attactgcag gtcgacgacc cagctttctt gtacaaagtg gttcgataat tcttaattaa
     6001 ctagttctag agcggccgcc accgcggtgg agctcgaatt tccccgatcg ttcaaacatt
     6061 tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa
     6121 tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg
     6181 agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa
     6241 atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg
     6301 gaattcgtaa tcatggtcat agctgtttcc tgcctagtcg gagcggccgc gtccgtctag
     6361 ctgatagtga ccttaggcga cttttgaacg cgcaataatg gtttctgacg tatgtgctta
     6421 gctcattaaa ctccagaaac ccgcggctga gtggctcctt caacgttgcg gttctgtcag
     6481 ttccaaacgt aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt gactccctta
     6541 attctccgct catgatcaga ttgtcgtttc ccgccttcag tttaaactat cagtgtttga
     6601 caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt
     6661 taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg
     6721 ttccccagat ctggcgccgg ccagcgagac gagcaagatt ggccgccgcc cgaaacgatc
     6781 cgacagcgcg cccagcacag gtgcgcaggc aaattgcacc aacgcataca gcgccagcag
     6841 aatgccatag tgggcggtga cgtcgttcga gtgaaccaga tcgcgcagga ggcccggcag
     6901 caccggcata atcaggccga tgccgacagc gtcgagcgcg acagtgctca gaattacgat
     6961 caggggtatg ttgggtttca cgtctggcct ccggaccagc ctccgctggt ccgattgaac
     7021 gcgcggattc tttatcactg ataagttggt ggacatatta tgtttatcag tgataaagtg
     7081 tcaagcatga caaagttgca gccgaataca gtgatccgtg ccgccctgga cctgttgaac
     7141 gaggtcggcg tagacggtct gacgacacgc aaactggcgg aacggttggg ggttcagcag
     7201 ccggcgcttt actggcactt caggaacaag cgggcgctgc tcgacgcact ggccgaagcc
     7261 atgctggcgg agaatcatac gcattcggtg ccgagagccg acgacgactg gcgctcattt
     7321 ctgatcggga atgcccgcag cttcaggcag gcgctgctcg cctaccgcga tggcgcgcgc
     7381 atccatgccg gcacgcgacc gggcgcaccg cagatggaaa cggccgacgc gcagcttcgc
     7441 ttcctctgcg aggcgggttt ttcggccggg gacgccgtca atgcgctgat gacaatcagc
     7501 tacttcactg ttggggccgt gcttgaggag caggccggcg acagcgatgc cggcgagcgc
     7561 ggcggcaccg ttgaacaggc tccgctctcg ccgctgttgc gggccgcgat agacgccttc
     7621 gacgaagccg gtccggacgc agcgttcgag cagggactcg cggtgattgt cgatggattg
     7681 gcgaaaagga ggctcgttgt caggaacgtt gaaggaccga gaaagggtga cgattgatca
     7741 ggaccgctgc cggagcgcaa cccactcact acagcagagc catgtagaca acatcccctc
     7801 cccctttcca ccgcgtcaga cgcccgtagc agcccgctac gggctttttc atgccctgcc
     7861 ctagcgtcca agcctcacgg ccgcgctcgg cctctctggc ggccttctgg cgctcttccg
     7921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8101 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8281 cgcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     8341 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     8401 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     8461 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     8521 tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga aaccaagcca
     8581 accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg aagagcgatt
     8641 gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc cgtcggccag
     8701 ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc ccgcatcaat
     8761 ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga cccgcgcacg
     8821 gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga gaagcaggac
     8881 gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg acttttttag
     8941 ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc gctccatcaa
     9001 gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca agaccgagcg
     9061 cctttccgac gctcaccggg ctggttgccc tcgccgctgg gctggcggcc gtctatggcc
     9121 ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag acaccgcggc cggccgccgg
     9181 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac
     9241 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg
     9301 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt
     9361 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg
     9421 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac
     9481 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt
     9541 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa
     9601 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg
     9661 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc
     9721 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc
     9781 agtccttgcc attgccggga tcggggcagt aacgggatgg gcgatcagcc cgagcgcgac
     9841 gcccggaagc attgacgtgc cgcaggtgct ggcatcgaca ttcagcgacc aggtgccggg
     9901 cagtgagggc ggcggcctgg gtggcggcct gcccttcact tcggccgtcg gggcattcac
     9961 ggacttcatg gcggggccgg caatttttac cttgggcatt cttggcatag tggtcgcggg
    10021 tgccgtgctc gtgttcgggg gtgcgataaa cccagcgaac catttgaggt gataggtaag
    10081 attataccga ggtatgaaaa cgagaattgg acctttacag aattactcta tgaagcgcca
    10141 tatttaaaaa gctaccaaga cgaagaggat gaagaggatg aggaggcaga ttgccttgaa
    10201 tatattgaca atactgataa gataatatat cttttatata gaagatatcg ccgtatgtaa
    10261 ggatttcagg gggcaaggca taggcagcgc gcttatcaat atatctatag aatgggcaaa
    10321 gcataaaaac ttgcatggac taatgcttga aacccaggac aataacctta tagcttgtaa
    10381 attctatcat aattgggtaa tgactccaac ttattgatag tgttttatgt tcagataatg
    10441 cccgatgact ttgtcatgca gctccaccga ttttgagaac gacagcgact tccgtcccag
    10501 ccgtgccagg tgctgcctca gattcaggtt atgccgctca attcgctgcg tatatcgctt
    10561 gctgattacg tgcagctttc ccttcaggcg ggattcatac agcggccagc catccgtcat
    10621 ccatatcacc acgtcaaagg gtgacagcag gctcataaga cgccccagcg tcgccatagt
    10681 gcgttcaccg aatacgtgcg caacaaccgt cttccggaga ctgtcatacg cgtaaaacag
    10741 ccagcgctgg cgcgatttag ccccgacata gccccactgt tcgtccattt ccgcgcagac
    10801 gatgacgtca ctgcccggct gtatgcgcga ggttaccgac tgcggcctga gttttttaag
    10861 tgacgtaaaa tcgtgttgag gccaacgccc ataatgcggg ctgttgcccg gcatccaacg
    10921 ccattcatgg ccatatcaat gattttctgg tgcgtaccgg gttgagaagc ggtgtaagtg
    10981 aactgcagtt gccatgtttt acggcagtga gagcagagat agcgctgatg tccggcggtg
    11041 cttttgccgt tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca
    11101 gaagccactg gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc
    11161 acccataatt gtggtttcaa aatcggctcc gtcgatacta tgttatacgc caactttgaa
    11221 aacaactttg aaaaagctgt tttctggtat ttaaggtttt agaatgcaag gaacagtgaa
    11281 ttggagttcg tcttgttata attagcttct tggggtatct ttaaatactg tagaaaagag
    11341 gaaggaaata ataaatggct aaaatgagaa tatcaccgga attgaaaaaa ctgatcgaaa
    11401 aataccgctg cgtaaaagat acggaaggaa tgtctcctgc taaggtatat aagctggtgg
    11461 gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg accacctatg
    11521 atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct gttccaaagg
    11581 tcctgcactt tgaacggcat gatggctgga gcaatctgct catgagtgag gccgatggcg
    11641 tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc gagctgtatg
    11701 cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat acgaatagct
    11761 tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc gatgtggatt
    11821 gcgaaaactg ggaagaagac actccattta aagatccgcg cgagctgtat gattttttaa
    11881 agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga gacagcaaca
    11941 tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc ggcagggcgg
    12001 acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc ggggaagaac
    12061 agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag aaaataaaat
    12121 attatatttt actggatgaa ttgttttagt acctagatgt ggcgcaacga tgccggcgac
    12181 aagcaggagc gcaccgactt cttccgcatc aagtgttttg gctctcaggc cgaggcccac
    12241 ggcaagtatt tgggcaaggg gtcgctggta ttcgtgcagg gcaagattcg gaataccaag
    12301 tacgagaagg acggccagac ggtctacggg accgacttca ttgccgataa ggtggattat
    12361 ctggacacca aggcaccagg cgggtcaaat caggaataag ggcacattgc cccggcgtga
    12421 gtcggggcaa tcccgcaagg agggtgaatg aatcggacgt ttgaccggaa ggcatacagg
    12481 caagaactga tcgacgcggg gttttccgcc gaggatgccg aaaccatcgc aagccgcacc
    12541 gtcatgcgtg cgccccgcga aaccttccag tccgtcggct cgatggtcca gcaagctacg
    12601 gccaagatcg agcgcgacag cgtgcaactg gctccccctg ccctgcccgc gccatcggcc
    12661 gccgtggagc gttcgcgtcg tctcgaacag gaggcggcag gtttggcgaa gtcgatgacc
    12721 atcgacacgc gaggaactat gacgaccaag aagcgaaaaa ccgccggcga ggacctggca
    12781 aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac acacgaagca gcagatcaag
    12841 gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg acacgatgcg agcgatgcca
    12901 aacgacacgg cccgctctgc cctgttcacc acgcgcaaca agaaaatccc gcgcgaggcg
    12961 ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga agatcaccta caccggcgtc
    13021 gagctgcggg ccgacgatga cgaactggtg tggcagcagg tgttggagta cgcgaagcgc
    13081 acccctatcg gcgagccgat caccttcacg ttctacgagc tttgccagga cctgggctgg
    13141 tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc tgtcgcgcct acaggcgacg
    13201 gcgatgggct tcacgtccga ccgcgttggg cacctggaat cggtgtcgct gctgcaccgc
    13261 ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc aggtcctgat cgacgaggaa
    13321 atcgtcgtgc tgtttgctgg cgaccactac acgaaattca tatgggagaa gtaccgcaag
    13381 ctgtcgccga cggcccgacg gatgttcgac tatttcagct cgcaccggga gccgtacccg
    13441 ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt ccacccgcgt gaagaagtgg
    13501 cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca gcggcctggt ggaacacgcc
    13561 tgggtcaatg atgacctggt gcattgcaaa cgctagggcc ttgtggggtc agttccggct
    13621 gggggttcag cagccagcgc tttactggca tttcaggaac aagcgggcac tgctcgacgc
    13681 acttgcttcg ctcagtatcg ctcgggacgc acggcgcgct ctacgaactg ccgataaaca
    13741 gaggattaaa attgacaatt gtgattaagg ctcagattcg acggcttgga gcggccgacg
    13801 tgcaggattt ccgcgagatc cgattgtcgg ccctgaagaa agctccagag atgttcgggt
    13861 ccgtttacga gcacgaggag aaaaagccca tggaggcgtt cgctgaacgg ttgcgagatg
    13921 ccgtggcatt cggcgcctac atcgacggcg agatcattgg gctgtcggtc ttcaaacagg
    13981 aggacggccc caaggacgct cacaaggcgc atctgtccgg cgttttcgtg gagcccgaac
    14041 agcgaggccg aggggtcgcc ggtatgctgc tgcgggcgtt gccggcgggt ttattgctcg
    14101 tgatgatcgt ccgacagatt ccaacgggaa tctggtggat gcgcatcttc atcctcggcg
    14161 cacttaatat ttcgctattc tggagcttgt tgtttatttc ggtctaccgc ctgccgggcg
    14221 gggtcgcggc gacggtaggc gctgtgcagc cgctgatggt cgtgttcatc tctgccgctc
    14281 tgctaggtag cccgatacga ttgatggcgg tcctgggggc tatttgcgga actgcgggcg
    14341 tggcgctgtt ggtgttgaca ccaaacgcag cgctagatcc tgtcggcgtc gcagcgggcc
    14401 tggcgggggc ggtttccatg gcgttcggaa ccgtgctgac ccgcaagtgg caacctcccg
    14461 tgcctctgct cacctttacc gcctggcaac tggcggccgg aggacttctg ctcgttccag
    14521 tagctttagt gtttgatccg ccaatcccga tgcctacagg aaccaatgtt ctcggcctgg
    14581 cgtggctcgg cctgatcgga gcgggtttaa cctacttcct ttggttccgg gggatctcgc
    14641 gactcgaacc tacagttgtt tccttactgg gctttctcag ccccagatct ggggtcgatc
    14701 agccggggat gcatcaggcc gacagtcgga acttcgggtc cccgacctgt accattcggt
    14761 gagcaatgga taggggagtt gatatcgtca acgttcactt ctaaagaaat agcgccactc
    14821 agcttcctca gcggctttat ccagcgattt cctattatgt cggcatagtt ctcaagatcg
    14881 acagcctgtc acggttaagc gagaaatgaa taagaaggct gataattcgg atctctgcga
    14941 gggagatgat atttgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac
    15001 cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc
    15061 atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg
    15121 actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt
    15181 tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat
    15241 aacacattgc ggacgttttt aatgtactgg ggtggttttt cttttcacca gtgagacggg
    15301 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
    15361 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt ccgaaatcgg caaaatccct
    15421 tataaatcaa aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt
    15481 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
    15541 ggcccactac gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca
    15601 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
    15661 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ccattcaggc tgcgcaactg
    15721 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
    15781 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
    15841 gacggccagt gaattaattc ccgatctagt aacatagatg acaccgcgcg cgataattta
    15901 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
    15961 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
    16021 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
    16081 cttaagaaac tttattgcca aatgtttgaa cgatcgggga aattcgagct cggtaccccc
    16141 tcctgggcac ggtcgtacgc gatccgccgc ggttccacct caccattgcg ggtatgccag
    16201 ttgagttcga tgagttcccg taccagctcc aactcgggcg atgccacgcg cactccgttc
    16261 agatctcggt gacgggcagg accggacggg gcggtaccgg caggctgaag tccagctgcc
    16321 agaaacccac gtcatgccag ttcccgtgct tgaagccggc cgcccgcagc atgccgcggg
    16381 gggcatatcc gagcgcctcg tgcatgcgca cgctcgggtc gttgggcagc ccgatgacag
    16441 cgaccacgct cttgaagccc tgtgcctcca gggacttcag caggtgggtg tagagcgtgg
    16501 agcccagtcc cgtccgctgg tggcgggggg agacgtacac ggtcgactcg gccgtccagt
    16561 cgtaggcgtt gcgtgccttc caggggcccg cgtaggcgat gccggcgacc tcgccgtcca
    16621 cctcggcgac gagccaggga tagcgctccc gcagacggac gaggtcgtcc gtccactcct
    16681 gcggttcctg cggctcggta cggaagttga ccgtgcttgt ctcgatgtag tggttgacga
    16741 tggtgcagac cgccggcatg tccgcctcgg tggcacggcg gatgtcggcc gggcgtcgtt
    16801 ctgggctcat ggatcctcta gagtcccccg tgttctctcc aaatgaaatg aacttcctta
    16861 tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg tcatccctta cgtcagtgga
    16921 gatatcacat caatccactt gctttgaaga cgtggttgga acgtcttctt tttccacgat
    16981 gctcctcgtg ggtgggggtc catctttggg accactgtcg gcagaggcat cttcaacgat
    17041 ggcctttcct ttatcgcaat gatggcattt gtaggagcca ccttcctttt ccactatctt
    17101 cacaataaag tgacagatag ctgggcaatg gaatccgagg aggtttccgg atattaccct
    17161 ttgttgaaaa gtctcaattg ccctttggtc ttctgagact gtatctttga tatttttgga
    17221 gtagacaagt gtgtcgtgct ccaccatgtt gacgaagatt ttcttcttgt cattgagtcg
    17281 taagagactc tgtatgaact gttcgccagt ctttacggcg agttctgtta ggtcctctat
    17341 ttgaatcttt gactccatgg cctttgattc agtgggaact acctttttag agactccaat
    17401 ctctattact tgccttggtt tgtgaagcaa gccttgaatc gtccatactg gaatagtact
    17461 tctgatcttg agaaatatat ctttctctgt gttcttgatg cagttagtcc tgaatctttt
    17521 gactgcatct ttaaccttct tgggaaggta tttgatttcc tggagattat tgctcgggta
    17581 gatcgtcttg atgagacctg ctgcgtaagc ctctctaacc atctgtgggt tagcattctt
    17641 tctgaaattg aaaaggctaa tctggggacc tgcaggggca ataccgccag ggttttccca
    17701 gtcacgacgt tgtaaaacga cggccagtgc caagctttcg acgagtcagt aataaacggc
    17761 gtcaaagtgg ttgcagccgg cacacacgag tcgtgtttat caactcaaag cacaaatact
    17821 tttcctcaac ctaaaaataa ggcaattagc caaaaacaac tttgcgtgta aacaacgctc
    17881 aatacacgtg tcattttatt attagctatt gcttcaccgc cttagctttc tcgtgaccta
    17941 gtcgtcctcg tcttttcttc ttcttcttct ataaaacaat acccaaagag ctcttcttct
    18001 tcacaattca gatttcaatt tctcaaaatc ttaaaaactt tctctcaatt ctctctaccg
    18061 tgatcaaggt aaatttctgt gttccttatt ctctcaaaat cttcgatttt gttttcgttc
    18121 gatcccaatt tcgtatatgt tctttggttt agattctgtt aatcttagat cgaagacgat
    18181 tttctgggtt tgatcgttag atatcatctt aattctcgat tagggtttca tagatatcat
    18241 ccgatttgtt caaataattt gagttttgtc gaataattac tcttcgattt gtgatttcta
    18301 tctagatctg gtgttagttt ctagtttgtg cgatcg
//
