LOCUS       sequence-202762-       18337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr8g060730 (Fbl1-like). 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: pSC34
FEATURES             Location/Qualifiers
     source          1..18337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    26..50
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             72..119
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             126..146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(171..1607,1797..4460)
                     /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          1608..1796
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4461..4508
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4519..4771
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4816..4836
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5191..5266
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5278..5302)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5780..5855
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5867..5891)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5910..5934)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      5998..6250
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6268..6284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6268..6284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6550..6574
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7039..7689
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7877..7894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8128..8147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8370..8479
                     /note="incP origin of transfer"
     CDS             8512..8883
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      9010..9725
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10348..11115
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11307..12101
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12400..13548
                     /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    15144..15168
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15415..15436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15627..15646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15786..15802
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15815..16067)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16211..16762)
                     /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(16783..17128)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16820..16840)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17664..17680
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccctcgaggc gcgccaagct atcaaacaag tttgtacaaa aaagcaggct ccgaattcgc
       61 ccttcaccat ggattacaag gatgatgatg ataaggatta caaggatgat gatgataaga
      121 tggctccaaa gaagaagaga aaggttggaa tccacggagt tccagctgct gataagaagt
      181 actctatcgg acttgacatc ggaaccaact ctgttggatg ggctgttatc accgatgagt
      241 acaaggttcc atctaagaag ttcaaggttc ttggaaacac cgatagacac tctatcaaga
      301 agaaccttat cggtgctctt cttttcgatt ctggagagac cgctgaggct accagattga
      361 agagaaccgc tagaagaaga tacaccagaa gaaagaacag aatctgctac cttcaggaaa
      421 tcttctctaa cgagatggct aaggttgatg attctttctt ccacagactt gaggagtctt
      481 tccttgttga ggaggataag aagcacgaga gacacccaat cttcggaaac atcgttgatg
      541 aggttgctta ccacgagaag tacccaacca tctaccacct tagaaagaag ttggttgatt
      601 ctaccgataa ggctgatctt agacttatct accttgctct tgctcacatg atcaagttca
      661 gaggacactt ccttatcgag ggagacctta acccagataa ctctgatgtt gataagttgt
      721 tcatccagct tgttcagacc tacaaccagc ttttcgagga gaacccaatc aacgcttctg
      781 gagttgatgc taaggctatc ctttctgcta gactttctaa gtctcgtaga cttgagaacc
      841 ttatcgctca gcttccagga gagaagaaga acggactttt cggaaacctt atcgctcttt
      901 ctcttggact taccccaaac ttcaagtcta acttcgatct tgctgaggat gctaagttgc
      961 agctttctaa ggatacctac gatgatgatc ttgataacct tcttgctcag atcggagatc
     1021 agtacgctga tcttttcctt gctgctaaga acctttctga tgctatcctt ctttctgaca
     1081 tccttagagt taacaccgag atcaccaagg ctccactttc tgcttctatg atcaagagat
     1141 acgatgagca ccaccaggat cttacccttt tgaaggctct tgttagacag cagcttccag
     1201 agaagtacaa ggaaatcttc ttcgatcagt ctaagaacgg atacgctgga tacatcgatg
     1261 gaggagcttc tcaggaggag ttctacaagt tcatcaagcc aatccttgag aagatggatg
     1321 gaaccgagga gcttcttgtt aagttgaaca gagaggatct tcttagaaag cagagaacct
     1381 tcgataacgg atctatccca caccagatcc accttggaga gcttcacgct atccttcgta
     1441 gacaggagga tttctaccca ttcttgaagg ataacagaga gaagatcgag aagatcctta
     1501 ccttcagaat cccatactac gttggaccac ttgctagagg aaactctcgt ttcgcttgga
     1561 tgaccagaaa gtctgaggag accatcaccc cttggaactt cgaggaggta agtttctgct
     1621 tctacctttg atatatatat aataattatc attaattagt agtaatataa tatttcaaat
     1681 atttttttca aaataaaaga atgtagtata tagcaattgc ttttctgtag tttataagtg
     1741 tgtatatttt aatttataac ttttctaata tatgaccaaa atttgttgat gtgcaggttg
     1801 ttgataaggg agcttctgct cagtctttca tcgagagaat gaccaacttc gataagaacc
     1861 ttccaaacga gaaggttctt ccaaagcact ctcttcttta cgagtacttc accgtttaca
     1921 acgagcttac caaggttaag tacgttaccg agggaatgag aaagccagct ttcctttctg
     1981 gagagcagaa gaaggctatc gttgatcttc ttttcaagac caacagaaag gttaccgtta
     2041 agcagttgaa ggaggattac ttcaagaaga tcgagtgctt cgattctgtt gaaatctctg
     2101 gagttgagga tagattcaac gcttctcttg gaacctacca cgatcttttg aagatcatca
     2161 aggataagga tttccttgat aacgaggaga acgaggacat ccttgaggac atcgttctta
     2221 cccttaccct tttcgaggat agagagatga tcgaggagag actcaagacc tacgctcacc
     2281 ttttcgatga taaggttatg aagcagttga agagaagaag atacaccgga tggggtagac
     2341 tttctcgtaa gttgatcaac ggaatcagag ataagcagtc tggaaagacc atccttgatt
     2401 tcttgaagtc tgatggattc gctaacagaa acttcatgca gcttatccac gatgattctc
     2461 ttaccttcaa ggaggacatc cagaaggctc aggtttctgg acagggagat tctcttcacg
     2521 agcacatcgc taaccttgct ggatctccag ctatcaagaa gggaatcctt cagaccgtta
     2581 aggttgttga tgagcttgtt aaggttatgg gtagacacaa gccagagaac atcgttatcg
     2641 agatggctag agagaaccag accacccaga agggacagaa gaactctcgt gagagaatga
     2701 agagaatcga ggagggaatc aaggagcttg gatctcaaat cttgaaggag cacccagttg
     2761 agaacaccca gcttcagaac gagaagttgt acctttacta ccttcagaac ggaagagata
     2821 tgtacgttga tcaggagctt gacatcaaca gactttctga ttacgatgtt gatcacatcg
     2881 ttccacagtc tttcttgaag gatgattcta tcgataacaa ggttcttacc cgttctgata
     2941 agaacagagg aaagtctgat aacgttccat ctgaggaggt tgttaagaag atgaagaact
     3001 actggagaca gcttcttaac gctaagttga tcacccagag aaagttcgat aaccttacca
     3061 aggctgagag aggaggactt tctgagcttg ataaggctgg attcatcaag agacagcttg
     3121 ttgagaccag acagatcacc aagcacgttg ctcagatcct tgattctcgt atgaacacca
     3181 agtacgatga gaacgataag ttgatcagag aggttaaggt tatcaccttg aagtctaagt
     3241 tggtttctga tttcagaaag gatttccagt tctacaaggt tagagagatc aacaactacc
     3301 accacgctca cgatgcttac cttaacgctg ttgttggaac cgctcttatc aagaagtacc
     3361 caaagttgga gtctgagttc gtttacggag attacaaggt ttacgatgtt agaaagatga
     3421 tcgctaagtc tgagcaggag atcggaaagg ctaccgctaa gtacttcttc tactctaaca
     3481 tcatgaactt cttcaagacc gagatcaccc ttgctaacgg agagatcaga aagagaccac
     3541 ttatcgagac caacggagag accggagaga tcgtttggga taagggaaga gatttcgcta
     3601 ccgttagaaa ggttctttct atgccacagg ttaacatcgt taagaaaacc gaggttcaga
     3661 ccggaggatt ctctaaggag tctatccttc caaagagaaa ctctgataag ttgatcgcta
     3721 gaaagaagga ttgggaccca aagaagtacg gaggattcga ttctccaacc gttgcttact
     3781 ctgttcttgt tgttgctaag gttgagaagg gaaagtctaa gaagttgaag tctgttaagg
     3841 agcttcttgg aatcaccatc atggagcgtt cttctttcga gaagaaccca atcgatttcc
     3901 ttgaggctaa gggatacaag gaggttaaga aggatcttat catcaagttg ccaaagtact
     3961 ctcttttcga gcttgagaac ggaagaaaga gaatgcttgc ttctgctgga gagcttcaga
     4021 agggaaacga gcttgctctt ccatctaagt acgttaactt cctttacctt gcttctcact
     4081 acgagaagtt gaagggatct ccagaggata acgagcagaa gcagcttttc gttgagcagc
     4141 acaagcacta ccttgatgag atcatcgagc aaatctctga gttctctaag agagttatcc
     4201 ttgctgatgc taaccttgat aaggttcttt ctgcttacaa caagcacaga gataagccaa
     4261 tcagagagca ggctgagaac atcatccacc ttttcaccct taccaacctt ggtgctccag
     4321 ctgctttcaa gtacttcgat accaccatcg atagaaaaag atacacctct accaaggagg
     4381 ttcttgatgc tacccttatc caccagtcta tcaccggact ttacgagacc agaatcgatc
     4441 tttctcagct tggaggagat aagagaccag ctgctaccaa gaaggctgga caggctaaga
     4501 agaagaagtg agacgtccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     4561 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     4621 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     4681 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     4741 tcgcgcgcgg tgtcatctat gttactagat cgggaattga tcccccctcg acagcttccg
     4801 gaaagggcga attcgcaact ttgtatacaa aagttgccga gctcgcggcc gcatgttgtt
     4861 gttaccagaa agtaaataaa tgttcaatct ctgatgttct caagtaagtg agttttattg
     4921 ggaataatat taacttatgt tcttcttgca tttgatttct ttgccgctct cttcttctat
     4981 cttaaatctg tgtatactat ttcactattg ggctttttat tagtctataa tgggactcaa
     5041 aataaggctt tggcccacat caaaaagata agtcacaaat caaaactaaa ttcagagtct
     5101 tttctcccac atcggtcact gtactcattt tgtgtttgtt tatatattac acgaaccgat
     5161 ctttggtacg caactggaag cttctcgaag gttttagagc tagaaatagc aagttaaaat
     5221 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctagacc
     5281 cagctttctt gtacaaagtt ggcattacca tggcttcgtt gaacaacgga aactcgactt
     5341 gccttccgca caatacatca tttcttctta gctttttttc ttcttcttcg ttcatacagt
     5401 ttttttttgt ttatcagctt acattttctt gaaccgtagc tttcgttttc ttctttttaa
     5461 ctttccattc ggagtttttg tatcttgttt catagtttgt cccaggatta gaatgattag
     5521 gcatcgaacc ttcaagaatt tgattgaata aaacatcttc attcttaaga tatgaagata
     5581 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag
     5641 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca
     5701 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattga
     5761 gtaaacctgc aatattccag ttttagagct agaaatagca agttaaaata aggctagtcc
     5821 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg
     5881 tacaaagttg gcattactgc aggtcgacga cccagctttc ttgtacaaag tggttcgata
     5941 attcttaatt aactagttct agagcggccg ccaccgcggt ggagctcgaa tttccccgat
     6001 cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg
     6061 attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg
     6121 acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg
     6181 atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg
     6241 ttactagatc gggaattcgt aatcatggtc atagctgttt cctgcctagt cggagcggcc
     6301 gcgtccgtct agctgatagt gaccttaggc gacttttgaa cgcgcaataa tggtttctga
     6361 cgtatgtgct tagctcatta aactccagaa acccgcggct gagtggctcc ttcaacgttg
     6421 cggttctgtc agttccaaac gtaaaacggc ttgtcccgcg tcatcggcgg gggtcataac
     6481 gtgactccct taattctccg ctcatgatca gattgtcgtt tcccgccttc agtttaaact
     6541 atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     6601 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
     6661 ccaaccacag ggttccccag atctggcgcc ggccagcgag acgagcaaga ttggccgccg
     6721 cccgaaacga tccgacagcg cgcccagcac aggtgcgcag gcaaattgca ccaacgcata
     6781 cagcgccagc agaatgccat agtgggcggt gacgtcgttc gagtgaacca gatcgcgcag
     6841 gaggcccggc agcaccggca taatcaggcc gatgccgaca gcgtcgagcg cgacagtgct
     6901 cagaattacg atcaggggta tgttgggttt cacgtctggc ctccggacca gcctccgctg
     6961 gtccgattga acgcgcggat tctttatcac tgataagttg gtggacatat tatgtttatc
     7021 agtgataaag tgtcaagcat gacaaagttg cagccgaata cagtgatccg tgccgccctg
     7081 gacctgttga acgaggtcgg cgtagacggt ctgacgacac gcaaactggc ggaacggttg
     7141 ggggttcagc agccggcgct ttactggcac ttcaggaaca agcgggcgct gctcgacgca
     7201 ctggccgaag ccatgctggc ggagaatcat acgcattcgg tgccgagagc cgacgacgac
     7261 tggcgctcat ttctgatcgg gaatgcccgc agcttcaggc aggcgctgct cgcctaccgc
     7321 gatggcgcgc gcatccatgc cggcacgcga ccgggcgcac cgcagatgga aacggccgac
     7381 gcgcagcttc gcttcctctg cgaggcgggt ttttcggccg gggacgccgt caatgcgctg
     7441 atgacaatca gctacttcac tgttggggcc gtgcttgagg agcaggccgg cgacagcgat
     7501 gccggcgagc gcggcggcac cgttgaacag gctccgctct cgccgctgtt gcgggccgcg
     7561 atagacgcct tcgacgaagc cggtccggac gcagcgttcg agcagggact cgcggtgatt
     7621 gtcgatggat tggcgaaaag gaggctcgtt gtcaggaacg ttgaaggacc gagaaagggt
     7681 gacgattgat caggaccgct gccggagcgc aacccactca ctacagcaga gccatgtaga
     7741 caacatcccc tccccctttc caccgcgtca gacgcccgta gcagcccgct acgggctttt
     7801 tcatgccctg ccctagcgtc caagcctcac ggccgcgctc ggcctctctg gcggccttct
     7861 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7921 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7981 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8041 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8101 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8161 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8221 cgggaagcgt ggcgcttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     8281 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     8341 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     8401 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     8461 gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgat
     8521 gaaaccaagc caaccaggaa gggcagccca cctatcaagg tgtactgcct tccagacgaa
     8581 cgaagagcga ttgaggaaaa ggcggcggcg gccggcatga gcctgtcggc ctacctgctg
     8641 gccgtcggcc agggctacaa aatcacgggc gtcgtggact atgagcacgt ccgcgagctg
     8701 gcccgcatca atggcgacct gggccgcctg ggcggcctgc tgaaactctg gctcaccgac
     8761 gacccgcgca cggcgcggtt cggtgatgcc acgatcctcg ccctgctggc gaagatcgaa
     8821 gagaagcagg acgagcttgg caaggtcatg atgggcgtgg tccgcccgag ggcagagcca
     8881 tgactttttt agccgctaaa acggccgggg ggtgcgcgtg attgccaagc acgtccccat
     8941 gcgctccatc aagaagagcg acttcgcgga gctggtgaag tacatcaccg acgagcaagg
     9001 caagaccgag cgcctttccg acgctcaccg ggctggttgc cctcgccgct gggctggcgg
     9061 ccgtctatgg ccctgcaaac gcgccagaaa cgccgtcgaa gccgtgtgcg agacaccgcg
     9121 gccggccgcc ggcgttgtgg atacctcgcg gaaaacttgg ccctcactga cagatgaggg
     9181 gcggacgttg acacttgagg ggccgactca cccggcgcgg cgttgacaga tgaggggcag
     9241 gctcgatttc ggccggcgac gtggagctgg ccagcctcgc aaatcggcga aaacgcctga
     9301 ttttacgcga gtttcccaca gatgatgtgg acaagcctgg ggataagtgc cctgcggtat
     9361 tgacacttga ggggcgcgac tactgacaga tgaggggcgc gatccttgac acttgagggg
     9421 cagagtgctg acagatgagg ggcgcaccta ttgacatttg aggggctgtc cacaggcaga
     9481 aaatccagca tttgcaaggg tttccgcccg tttttcggcc accgctaacc tgtcttttaa
     9541 cctgctttta aaccaatatt tataaacctt gtttttaacc agggctgcgc cctgtgcgcg
     9601 tgaccgcgca cgccgaaggg gggtgccccc ccttctcgaa ccctcccggc ccgctaacgc
     9661 gggcctccca tccccccagg ggctgcgccc ctcggccgcg aacggcctca ccccaaaaat
     9721 ggcagcgctg gcagtccttg ccattgccgg gatcggggca gtaacgggat gggcgatcag
     9781 cccgagcgcg acgcccggaa gcattgacgt gccgcaggtg ctggcatcga cattcagcga
     9841 ccaggtgccg ggcagtgagg gcggcggcct gggtggcggc ctgcccttca cttcggccgt
     9901 cggggcattc acggacttca tggcggggcc ggcaattttt accttgggca ttcttggcat
     9961 agtggtcgcg ggtgccgtgc tcgtgttcgg gggtgcgata aacccagcga accatttgag
    10021 gtgataggta agattatacc gaggtatgaa aacgagaatt ggacctttac agaattactc
    10081 tatgaagcgc catatttaaa aagctaccaa gacgaagagg atgaagagga tgaggaggca
    10141 gattgccttg aatatattga caatactgat aagataatat atcttttata tagaagatat
    10201 cgccgtatgt aaggatttca gggggcaagg cataggcagc gcgcttatca atatatctat
    10261 agaatgggca aagcataaaa acttgcatgg actaatgctt gaaacccagg acaataacct
    10321 tatagcttgt aaattctatc ataattgggt aatgactcca acttattgat agtgttttat
    10381 gttcagataa tgcccgatga ctttgtcatg cagctccacc gattttgaga acgacagcga
    10441 cttccgtccc agccgtgcca ggtgctgcct cagattcagg ttatgccgct caattcgctg
    10501 cgtatatcgc ttgctgatta cgtgcagctt tcccttcagg cgggattcat acagcggcca
    10561 gccatccgtc atccatatca ccacgtcaaa gggtgacagc aggctcataa gacgccccag
    10621 cgtcgccata gtgcgttcac cgaatacgtg cgcaacaacc gtcttccgga gactgtcata
    10681 cgcgtaaaac agccagcgct ggcgcgattt agccccgaca tagccccact gttcgtccat
    10741 ttccgcgcag acgatgacgt cactgcccgg ctgtatgcgc gaggttaccg actgcggcct
    10801 gagtttttta agtgacgtaa aatcgtgttg aggccaacgc ccataatgcg ggctgttgcc
    10861 cggcatccaa cgccattcat ggccatatca atgattttct ggtgcgtacc gggttgagaa
    10921 gcggtgtaag tgaactgcag ttgccatgtt ttacggcagt gagagcagag atagcgctga
    10981 tgtccggcgg tgcttttgcc gttacgcacc accccgtcag tagctgaaca ggagggacag
    11041 ctgatagaaa cagaagccac tggagcacct caaaaacacc atcatacact aaatcagtaa
    11101 gttggcagca tcacccataa ttgtggtttc aaaatcggct ccgtcgatac tatgttatac
    11161 gccaactttg aaaacaactt tgaaaaagct gttttctggt atttaaggtt ttagaatgca
    11221 aggaacagtg aattggagtt cgtcttgtta taattagctt cttggggtat ctttaaatac
    11281 tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag aatatcaccg gaattgaaaa
    11341 aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct gctaaggtat
    11401 ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc cggtataaag
    11461 ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa ggaaagctgc
    11521 ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg ctcatgagtg
    11581 aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct gaaaagatta
    11641 tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg gattgtccct
    11701 atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat aacgatctgg
    11761 ccgatgtgga ttgcgaaaac tgggaagaag acactccatt taaagatccg cgcgagctgt
    11821 atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac ggcgacctgg
    11881 gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat cttgggagaa
    11941 gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgatc agggaggata
    12001 tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag cctgattggg
    12061 agaaaataaa atattatatt ttactggatg aattgtttta gtacctagat gtggcgcaac
    12121 gatgccggcg acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag
    12181 gccgaggccc acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt
    12241 cggaatacca agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat
    12301 aaggtggatt atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt
    12361 gccccggcgt gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg
    12421 aaggcataca ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc
    12481 gcaagccgca ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc
    12541 cagcaagcta cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc
    12601 gcgccatcgg ccgccgtgga gcgttcgcgt cgtctcgaac aggaggcggc aggtttggcg
    12661 aagtcgatga ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc
    12721 gaggacctgg caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag
    12781 cagcagatca aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg
    12841 cgagcgatgc caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc
    12901 ccgcgcgagg cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc
    12961 tacaccggcg tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag
    13021 tacgcgaagc gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag
    13081 gacctgggct ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc
    13141 ctacaggcga cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg
    13201 ctgctgcacc gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg
    13261 atcgacgagg aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag
    13321 aagtaccgca agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg
    13381 gagccgtacc cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc
    13441 gtgaagaagt ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg
    13501 gtggaacacg cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg
    13561 tcagttccgg ctgggggttc agcagccagc gctttactgg catttcagga acaagcgggc
    13621 actgctcgac gcacttgctt cgctcagtat cgctcgggac gcacggcgcg ctctacgaac
    13681 tgccgataaa cagaggatta aaattgacaa ttgtgattaa ggctcagatt cgacggcttg
    13741 gagcggccga cgtgcaggat ttccgcgaga tccgattgtc ggccctgaag aaagctccag
    13801 agatgttcgg gtccgtttac gagcacgagg agaaaaagcc catggaggcg ttcgctgaac
    13861 ggttgcgaga tgccgtggca ttcggcgcct acatcgacgg cgagatcatt gggctgtcgg
    13921 tcttcaaaca ggaggacggc cccaaggacg ctcacaaggc gcatctgtcc ggcgttttcg
    13981 tggagcccga acagcgaggc cgaggggtcg ccggtatgct gctgcgggcg ttgccggcgg
    14041 gtttattgct cgtgatgatc gtccgacaga ttccaacggg aatctggtgg atgcgcatct
    14101 tcatcctcgg cgcacttaat atttcgctat tctggagctt gttgtttatt tcggtctacc
    14161 gcctgccggg cggggtcgcg gcgacggtag gcgctgtgca gccgctgatg gtcgtgttca
    14221 tctctgccgc tctgctaggt agcccgatac gattgatggc ggtcctgggg gctatttgcg
    14281 gaactgcggg cgtggcgctg ttggtgttga caccaaacgc agcgctagat cctgtcggcg
    14341 tcgcagcggg cctggcgggg gcggtttcca tggcgttcgg aaccgtgctg acccgcaagt
    14401 ggcaacctcc cgtgcctctg ctcaccttta ccgcctggca actggcggcc ggaggacttc
    14461 tgctcgttcc agtagcttta gtgtttgatc cgccaatccc gatgcctaca ggaaccaatg
    14521 ttctcggcct ggcgtggctc ggcctgatcg gagcgggttt aacctacttc ctttggttcc
    14581 gggggatctc gcgactcgaa cctacagttg tttccttact gggctttctc agccccagat
    14641 ctggggtcga tcagccgggg atgcatcagg ccgacagtcg gaacttcggg tccccgacct
    14701 gtaccattcg gtgagcaatg gataggggag ttgatatcgt caacgttcac ttctaaagaa
    14761 atagcgccac tcagcttcct cagcggcttt atccagcgat ttcctattat gtcggcatag
    14821 ttctcaagat cgacagcctg tcacggttaa gcgagaaatg aataagaagg ctgataattc
    14881 ggatctctgc gagggagatg atatttgatc acaggcagca acgctctgtc atcgttacaa
    14941 tcaacatgct accctccgcg agatcatccg tgtttcaaac ccggcagctt agttgccgtt
    15001 cttccgaata gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc tctcccgctg
    15061 acgccgtccc ggactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg
    15121 tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt
    15181 agacaactta ataacacatt gcggacgttt ttaatgtact ggggtggttt ttcttttcac
    15241 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
    15301 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttccgaaatc
    15361 ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt
    15421 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
    15481 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
    15541 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
    15601 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgccattcag
    15661 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
    15721 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
    15781 acgttgtaaa acgacggcca gtgaattaat tcccgatcta gtaacataga tgacaccgcg
    15841 cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat
    15901 aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta
    15961 attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca
    16021 acaggattca atcttaagaa actttattgc caaatgtttg aacgatcggg gaaattcgag
    16081 ctcggtaccc cctcctgggc acggtcgtac gcgatccgcc gcggttccac ctcaccattg
    16141 cgggtatgcc agttgagttc gatgagttcc cgtaccagct ccaactcggg cgatgccacg
    16201 cgcactccgt tcagatctcg gtgacgggca ggaccggacg gggcggtacc ggcaggctga
    16261 agtccagctg ccagaaaccc acgtcatgcc agttcccgtg cttgaagccg gccgcccgca
    16321 gcatgccgcg gggggcatat ccgagcgcct cgtgcatgcg cacgctcggg tcgttgggca
    16381 gcccgatgac agcgaccacg ctcttgaagc cctgtgcctc cagggacttc agcaggtggg
    16441 tgtagagcgt ggagcccagt cccgtccgct ggtggcgggg ggagacgtac acggtcgact
    16501 cggccgtcca gtcgtaggcg ttgcgtgcct tccaggggcc cgcgtaggcg atgccggcga
    16561 cctcgccgtc cacctcggcg acgagccagg gatagcgctc ccgcagacgg acgaggtcgt
    16621 ccgtccactc ctgcggttcc tgcggctcgg tacggaagtt gaccgtgctt gtctcgatgt
    16681 agtggttgac gatggtgcag accgccggca tgtccgcctc ggtggcacgg cggatgtcgg
    16741 ccgggcgtcg ttctgggctc atggatcctc tagagtcccc cgtgttctct ccaaatgaaa
    16801 tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct
    16861 tacgtcagtg gagatatcac atcaatccac ttgctttgaa gacgtggttg gaacgtcttc
    16921 tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagaggc
    16981 atcttcaacg atggcctttc ctttatcgca atgatggcat ttgtaggagc caccttcctt
    17041 ttccactatc ttcacaataa agtgacagat agctgggcaa tggaatccga ggaggtttcc
    17101 ggatattacc ctttgttgaa aagtctcaat tgccctttgg tcttctgaga ctgtatcttt
    17161 gatatttttg gagtagacaa gtgtgtcgtg ctccaccatg ttgacgaaga ttttcttctt
    17221 gtcattgagt cgtaagagac tctgtatgaa ctgttcgcca gtctttacgg cgagttctgt
    17281 taggtcctct atttgaatct ttgactccat ggcctttgat tcagtgggaa ctaccttttt
    17341 agagactcca atctctatta cttgccttgg tttgtgaagc aagccttgaa tcgtccatac
    17401 tggaatagta cttctgatct tgagaaatat atctttctct gtgttcttga tgcagttagt
    17461 cctgaatctt ttgactgcat ctttaacctt cttgggaagg tatttgattt cctggagatt
    17521 attgctcggg tagatcgtct tgatgagacc tgctgcgtaa gcctctctaa ccatctgtgg
    17581 gttagcattc tttctgaaat tgaaaaggct aatctgggga cctgcagggg caataccgcc
    17641 agggttttcc cagtcacgac gttgtaaaac gacggccagt gccaagcttt cgacgagtca
    17701 gtaataaacg gcgtcaaagt ggttgcagcc ggcacacacg agtcgtgttt atcaactcaa
    17761 agcacaaata cttttcctca acctaaaaat aaggcaatta gccaaaaaca actttgcgtg
    17821 taaacaacgc tcaatacacg tgtcatttta ttattagcta ttgcttcacc gccttagctt
    17881 tctcgtgacc tagtcgtcct cgtcttttct tcttcttctt ctataaaaca atacccaaag
    17941 agctcttctt cttcacaatt cagatttcaa tttctcaaaa tcttaaaaac tttctctcaa
    18001 ttctctctac cgtgatcaag gtaaatttct gtgttcctta ttctctcaaa atcttcgatt
    18061 ttgttttcgt tcgatcccaa tttcgtatat gttctttggt ttagattctg ttaatcttag
    18121 atcgaagacg attttctggg tttgatcgtt agatatcatc ttaattctcg attagggttt
    18181 catagatatc atccgatttg ttcaaataat ttgagttttg tcgaataatt actcttcgat
    18241 ttgtgatttc tatctagatc tggtgttagt ttctagtttg tgcgatcgaa tttgtagatt
    18301 aatctgagtt tttctgatta acagggtacc gggcccc
//
