LOCUS       sequence-202709-       18336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g101040 (Mel1). Also 
            expresses Cas9 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: pSC26
FEATURES             Location/Qualifiers
     source          1..18336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    34..58
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             80..127
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             134..154
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(179..1615,1805..4468)
                     /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          1616..1804
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             4469..4516
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4527..4779
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4824..4844
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        5199..5274
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5286..5310)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        5787..5862
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(5874..5898)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5917..5941)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      6005..6257
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6275..6291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6275..6291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    6557..6581
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             7046..7696
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     complement(7884..7901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(8135..8154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            8377..8486
                     /note="incP origin of transfer"
     CDS             8519..8890
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      9017..9732
                     /label=oriV
                     /note="incP origin of replication"
     mobile_element  10355..11122
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     CDS             11314..12108
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             12407..13555
                     /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    15151..15175
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(15422..15443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15634..15653
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15793..15809
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(15822..16074)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(16218..16769)
                     /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(16790..17135)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(16827..16847)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     primer_bind     17671..17687
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgggcccccc ctcgaggcgc gccaagctat caaacaagtt tgtacaaaaa agcaggctcc
       61 gaattcgccc ttcaccatgg attacaagga tgatgatgat aaggattaca aggatgatga
      121 tgataagatg gctccaaaga agaagagaaa ggttggaatc cacggagttc cagctgctga
      181 taagaagtac tctatcggac ttgacatcgg aaccaactct gttggatggg ctgttatcac
      241 cgatgagtac aaggttccat ctaagaagtt caaggttctt ggaaacaccg atagacactc
      301 tatcaagaag aaccttatcg gtgctcttct tttcgattct ggagagaccg ctgaggctac
      361 cagattgaag agaaccgcta gaagaagata caccagaaga aagaacagaa tctgctacct
      421 tcaggaaatc ttctctaacg agatggctaa ggttgatgat tctttcttcc acagacttga
      481 ggagtctttc cttgttgagg aggataagaa gcacgagaga cacccaatct tcggaaacat
      541 cgttgatgag gttgcttacc acgagaagta cccaaccatc taccacctta gaaagaagtt
      601 ggttgattct accgataagg ctgatcttag acttatctac cttgctcttg ctcacatgat
      661 caagttcaga ggacacttcc ttatcgaggg agaccttaac ccagataact ctgatgttga
      721 taagttgttc atccagcttg ttcagaccta caaccagctt ttcgaggaga acccaatcaa
      781 cgcttctgga gttgatgcta aggctatcct ttctgctaga ctttctaagt ctcgtagact
      841 tgagaacctt atcgctcagc ttccaggaga gaagaagaac ggacttttcg gaaaccttat
      901 cgctctttct cttggactta ccccaaactt caagtctaac ttcgatcttg ctgaggatgc
      961 taagttgcag ctttctaagg atacctacga tgatgatctt gataaccttc ttgctcagat
     1021 cggagatcag tacgctgatc ttttccttgc tgctaagaac ctttctgatg ctatccttct
     1081 ttctgacatc cttagagtta acaccgagat caccaaggct ccactttctg cttctatgat
     1141 caagagatac gatgagcacc accaggatct tacccttttg aaggctcttg ttagacagca
     1201 gcttccagag aagtacaagg aaatcttctt cgatcagtct aagaacggat acgctggata
     1261 catcgatgga ggagcttctc aggaggagtt ctacaagttc atcaagccaa tccttgagaa
     1321 gatggatgga accgaggagc ttcttgttaa gttgaacaga gaggatcttc ttagaaagca
     1381 gagaaccttc gataacggat ctatcccaca ccagatccac cttggagagc ttcacgctat
     1441 ccttcgtaga caggaggatt tctacccatt cttgaaggat aacagagaga agatcgagaa
     1501 gatccttacc ttcagaatcc catactacgt tggaccactt gctagaggaa actctcgttt
     1561 cgcttggatg accagaaagt ctgaggagac catcacccct tggaacttcg aggaggtaag
     1621 tttctgcttc tacctttgat atatatataa taattatcat taattagtag taatataata
     1681 tttcaaatat ttttttcaaa ataaaagaat gtagtatata gcaattgctt ttctgtagtt
     1741 tataagtgtg tatattttaa tttataactt ttctaatata tgaccaaaat ttgttgatgt
     1801 gcaggttgtt gataagggag cttctgctca gtctttcatc gagagaatga ccaacttcga
     1861 taagaacctt ccaaacgaga aggttcttcc aaagcactct cttctttacg agtacttcac
     1921 cgtttacaac gagcttacca aggttaagta cgttaccgag ggaatgagaa agccagcttt
     1981 cctttctgga gagcagaaga aggctatcgt tgatcttctt ttcaagacca acagaaaggt
     2041 taccgttaag cagttgaagg aggattactt caagaagatc gagtgcttcg attctgttga
     2101 aatctctgga gttgaggata gattcaacgc ttctcttgga acctaccacg atcttttgaa
     2161 gatcatcaag gataaggatt tccttgataa cgaggagaac gaggacatcc ttgaggacat
     2221 cgttcttacc cttacccttt tcgaggatag agagatgatc gaggagagac tcaagaccta
     2281 cgctcacctt ttcgatgata aggttatgaa gcagttgaag agaagaagat acaccggatg
     2341 gggtagactt tctcgtaagt tgatcaacgg aatcagagat aagcagtctg gaaagaccat
     2401 ccttgatttc ttgaagtctg atggattcgc taacagaaac ttcatgcagc ttatccacga
     2461 tgattctctt accttcaagg aggacatcca gaaggctcag gtttctggac agggagattc
     2521 tcttcacgag cacatcgcta accttgctgg atctccagct atcaagaagg gaatccttca
     2581 gaccgttaag gttgttgatg agcttgttaa ggttatgggt agacacaagc cagagaacat
     2641 cgttatcgag atggctagag agaaccagac cacccagaag ggacagaaga actctcgtga
     2701 gagaatgaag agaatcgagg agggaatcaa ggagcttgga tctcaaatct tgaaggagca
     2761 cccagttgag aacacccagc ttcagaacga gaagttgtac ctttactacc ttcagaacgg
     2821 aagagatatg tacgttgatc aggagcttga catcaacaga ctttctgatt acgatgttga
     2881 tcacatcgtt ccacagtctt tcttgaagga tgattctatc gataacaagg ttcttacccg
     2941 ttctgataag aacagaggaa agtctgataa cgttccatct gaggaggttg ttaagaagat
     3001 gaagaactac tggagacagc ttcttaacgc taagttgatc acccagagaa agttcgataa
     3061 ccttaccaag gctgagagag gaggactttc tgagcttgat aaggctggat tcatcaagag
     3121 acagcttgtt gagaccagac agatcaccaa gcacgttgct cagatccttg attctcgtat
     3181 gaacaccaag tacgatgaga acgataagtt gatcagagag gttaaggtta tcaccttgaa
     3241 gtctaagttg gtttctgatt tcagaaagga tttccagttc tacaaggtta gagagatcaa
     3301 caactaccac cacgctcacg atgcttacct taacgctgtt gttggaaccg ctcttatcaa
     3361 gaagtaccca aagttggagt ctgagttcgt ttacggagat tacaaggttt acgatgttag
     3421 aaagatgatc gctaagtctg agcaggagat cggaaaggct accgctaagt acttcttcta
     3481 ctctaacatc atgaacttct tcaagaccga gatcaccctt gctaacggag agatcagaaa
     3541 gagaccactt atcgagacca acggagagac cggagagatc gtttgggata agggaagaga
     3601 tttcgctacc gttagaaagg ttctttctat gccacaggtt aacatcgtta agaaaaccga
     3661 ggttcagacc ggaggattct ctaaggagtc tatccttcca aagagaaact ctgataagtt
     3721 gatcgctaga aagaaggatt gggacccaaa gaagtacgga ggattcgatt ctccaaccgt
     3781 tgcttactct gttcttgttg ttgctaaggt tgagaaggga aagtctaaga agttgaagtc
     3841 tgttaaggag cttcttggaa tcaccatcat ggagcgttct tctttcgaga agaacccaat
     3901 cgatttcctt gaggctaagg gatacaagga ggttaagaag gatcttatca tcaagttgcc
     3961 aaagtactct cttttcgagc ttgagaacgg aagaaagaga atgcttgctt ctgctggaga
     4021 gcttcagaag ggaaacgagc ttgctcttcc atctaagtac gttaacttcc tttaccttgc
     4081 ttctcactac gagaagttga agggatctcc agaggataac gagcagaagc agcttttcgt
     4141 tgagcagcac aagcactacc ttgatgagat catcgagcaa atctctgagt tctctaagag
     4201 agttatcctt gctgatgcta accttgataa ggttctttct gcttacaaca agcacagaga
     4261 taagccaatc agagagcagg ctgagaacat catccacctt ttcaccctta ccaaccttgg
     4321 tgctccagct gctttcaagt acttcgatac caccatcgat agaaaaagat acacctctac
     4381 caaggaggtt cttgatgcta cccttatcca ccagtctatc accggacttt acgagaccag
     4441 aatcgatctt tctcagcttg gaggagataa gagaccagct gctaccaaga aggctggaca
     4501 ggctaagaag aagaagtgag acgtccgatc gttcaaacat ttggcaataa agtttcttaa
     4561 gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta
     4621 agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta
     4681 gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg
     4741 ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattgatc ccccctcgac
     4801 agcttccgga aagggcgaat tcgcaacttt gtatacaaaa gttgccgagc tcgcggccgc
     4861 atgttgttgt taccagaaag taaataaatg ttcaatctct gatgttctca agtaagtgag
     4921 ttttattggg aataatatta acttatgttc ttcttgcatt tgatttcttt gccgctctct
     4981 tcttctatct taaatctgtg tatactattt cactattggg ctttttatta gtctataatg
     5041 ggactcaaaa taaggctttg gcccacatca aaaagataag tcacaaatca aaactaaatt
     5101 cagagtcttt tctcccacat cggtcactgt actcattttg tgtttgttta tatattacac
     5161 gaaccgatct ttggtacggg acatttttat gcgtccacgt tttagagcta gaaatagcaa
     5221 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     5281 tctagaccca gctttcttgt acaaagttgg cattaccatg gcttcgttga acaacggaaa
     5341 ctcgacttgc cttccgcaca atacatcatt tcttcttagc tttttttctt cttcttcgtt
     5401 catacagttt ttttttgttt atcagcttac attttcttga accgtagctt tcgttttctt
     5461 ctttttaact ttccattcgg agtttttgta tcttgtttca tagtttgtcc caggattaga
     5521 atgattaggc atcgaacctt caagaatttg attgaataaa acatcttcat tcttaagata
     5581 tgaagataat cttcaaaagg cccctgggaa tctgaaagaa gagaagcagg cccatttata
     5641 tgggaaagaa caatagtatt tcttatatag gcccatttaa gttgaaaaca atcttcaaaa
     5701 gtcccacatc gcttagataa gaaaacgaag ctgagtttat atacagctag agtcgaagta
     5761 gtgattgtgg atgataatcc gatctcgttt tagagctaga aatagcaagt taaaataagg
     5821 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc tagacccagc
     5881 tttcttgtac aaagttggca ttactgcagg tcgacgaccc agctttcttg tacaaagtgg
     5941 ttcgataatt cttaattaac tagttctaga gcggccgcca ccgcggtgga gctcgaattt
     6001 ccccgatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct
     6061 tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta
     6121 atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta
     6181 atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc
     6241 atctatgtta ctagatcggg aattcgtaat catggtcata gctgtttcct gcctagtcgg
     6301 agcggccgcg tccgtctagc tgatagtgac cttaggcgac ttttgaacgc gcaataatgg
     6361 tttctgacgt atgtgcttag ctcattaaac tccagaaacc cgcggctgag tggctccttc
     6421 aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca tcggcggggg
     6481 tcataacgtg actcccttaa ttctccgctc atgatcagat tgtcgtttcc cgccttcagt
     6541 ttaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt
     6601 attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat
     6661 gtgcatgcca accacagggt tccccagatc tggcgccggc cagcgagacg agcaagattg
     6721 gccgccgccc gaaacgatcc gacagcgcgc ccagcacagg tgcgcaggca aattgcacca
     6781 acgcatacag cgccagcaga atgccatagt gggcggtgac gtcgttcgag tgaaccagat
     6841 cgcgcaggag gcccggcagc accggcataa tcaggccgat gccgacagcg tcgagcgcga
     6901 cagtgctcag aattacgatc aggggtatgt tgggtttcac gtctggcctc cggaccagcc
     6961 tccgctggtc cgattgaacg cgcggattct ttatcactga taagttggtg gacatattat
     7021 gtttatcagt gataaagtgt caagcatgac aaagttgcag ccgaatacag tgatccgtgc
     7081 cgccctggac ctgttgaacg aggtcggcgt agacggtctg acgacacgca aactggcgga
     7141 acggttgggg gttcagcagc cggcgcttta ctggcacttc aggaacaagc gggcgctgct
     7201 cgacgcactg gccgaagcca tgctggcgga gaatcatacg cattcggtgc cgagagccga
     7261 cgacgactgg cgctcatttc tgatcgggaa tgcccgcagc ttcaggcagg cgctgctcgc
     7321 ctaccgcgat ggcgcgcgca tccatgccgg cacgcgaccg ggcgcaccgc agatggaaac
     7381 ggccgacgcg cagcttcgct tcctctgcga ggcgggtttt tcggccgggg acgccgtcaa
     7441 tgcgctgatg acaatcagct acttcactgt tggggccgtg cttgaggagc aggccggcga
     7501 cagcgatgcc ggcgagcgcg gcggcaccgt tgaacaggct ccgctctcgc cgctgttgcg
     7561 ggccgcgata gacgccttcg acgaagccgg tccggacgca gcgttcgagc agggactcgc
     7621 ggtgattgtc gatggattgg cgaaaaggag gctcgttgtc aggaacgttg aaggaccgag
     7681 aaagggtgac gattgatcag gaccgctgcc ggagcgcaac ccactcacta cagcagagcc
     7741 atgtagacaa catcccctcc ccctttccac cgcgtcagac gcccgtagca gcccgctacg
     7801 ggctttttca tgccctgccc tagcgtccaa gcctcacggc cgcgctcggc ctctctggcg
     7861 gccttctggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     8041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     8101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     8161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     8221 ctcccttcgg gaagcgtggc gcttttccgc tgcataaccc tgcttcgggg tcattatagc
     8281 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     8341 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     8401 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     8461 gaatcctgct ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat
     8521 ggctgatgaa accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc
     8581 agacgaacga agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta
     8641 cctgctggcc gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg
     8701 cgagctggcc cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct
     8761 caccgacgac ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa
     8821 gatcgaagag aagcaggacg agcttggcaa ggtcatgatg ggcgtggtcc gcccgagggc
     8881 agagccatga cttttttagc cgctaaaacg gccggggggt gcgcgtgatt gccaagcacg
     8941 tccccatgcg ctccatcaag aagagcgact tcgcggagct ggtgaagtac atcaccgacg
     9001 agcaaggcaa gaccgagcgc ctttccgacg ctcaccgggc tggttgccct cgccgctggg
     9061 ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga
     9121 caccgcggcc ggccgccggc gttgtggata cctcgcggaa aacttggccc tcactgacag
     9181 atgaggggcg gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt tgacagatga
     9241 ggggcaggct cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa
     9301 cgcctgattt tacgcgagtt tcccacagat gatgtggaca agcctgggga taagtgccct
     9361 gcggtattga cacttgaggg gcgcgactac tgacagatga ggggcgcgat ccttgacact
     9421 tgaggggcag agtgctgaca gatgaggggc gcacctattg acatttgagg ggctgtccac
     9481 aggcagaaaa tccagcattt gcaagggttt ccgcccgttt ttcggccacc gctaacctgt
     9541 cttttaacct gcttttaaac caatatttat aaaccttgtt tttaaccagg gctgcgccct
     9601 gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc tcccggcccg
     9661 ctaacgcggg cctcccatcc ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc
     9721 caaaaatggc agcgctggca gtccttgcca ttgccgggat cggggcagta acgggatggg
     9781 cgatcagccc gagcgcgacg cccggaagca ttgacgtgcc gcaggtgctg gcatcgacat
     9841 tcagcgacca ggtgccgggc agtgagggcg gcggcctggg tggcggcctg cccttcactt
     9901 cggccgtcgg ggcattcacg gacttcatgg cggggccggc aatttttacc ttgggcattc
     9961 ttggcatagt ggtcgcgggt gccgtgctcg tgttcggggg tgcgataaac ccagcgaacc
    10021 atttgaggtg ataggtaaga ttataccgag gtatgaaaac gagaattgga cctttacaga
    10081 attactctat gaagcgccat atttaaaaag ctaccaagac gaagaggatg aagaggatga
    10141 ggaggcagat tgccttgaat atattgacaa tactgataag ataatatatc ttttatatag
    10201 aagatatcgc cgtatgtaag gatttcaggg ggcaaggcat aggcagcgcg cttatcaata
    10261 tatctataga atgggcaaag cataaaaact tgcatggact aatgcttgaa acccaggaca
    10321 ataaccttat agcttgtaaa ttctatcata attgggtaat gactccaact tattgatagt
    10381 gttttatgtt cagataatgc ccgatgactt tgtcatgcag ctccaccgat tttgagaacg
    10441 acagcgactt ccgtcccagc cgtgccaggt gctgcctcag attcaggtta tgccgctcaa
    10501 ttcgctgcgt atatcgcttg ctgattacgt gcagctttcc cttcaggcgg gattcataca
    10561 gcggccagcc atccgtcatc catatcacca cgtcaaaggg tgacagcagg ctcataagac
    10621 gccccagcgt cgccatagtg cgttcaccga atacgtgcgc aacaaccgtc ttccggagac
    10681 tgtcatacgc gtaaaacagc cagcgctggc gcgatttagc cccgacatag ccccactgtt
    10741 cgtccatttc cgcgcagacg atgacgtcac tgcccggctg tatgcgcgag gttaccgact
    10801 gcggcctgag ttttttaagt gacgtaaaat cgtgttgagg ccaacgccca taatgcgggc
    10861 tgttgcccgg catccaacgc cattcatggc catatcaatg attttctggt gcgtaccggg
    10921 ttgagaagcg gtgtaagtga actgcagttg ccatgtttta cggcagtgag agcagagata
    10981 gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga
    11041 gggacagctg atagaaacag aagccactgg agcacctcaa aaacaccatc atacactaaa
    11101 tcagtaagtt ggcagcatca cccataattg tggtttcaaa atcggctccg tcgatactat
    11161 gttatacgcc aactttgaaa acaactttga aaaagctgtt ttctggtatt taaggtttta
    11221 gaatgcaagg aacagtgaat tggagttcgt cttgttataa ttagcttctt ggggtatctt
    11281 taaatactgt agaaaagagg aaggaaataa taaatggcta aaatgagaat atcaccggaa
    11341 ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat gtctcctgct
    11401 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
    11461 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
    11521 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
    11581 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa
    11641 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
    11701 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
    11761 gatctggccg atgtggattg cgaaaactgg gaagaagaca ctccatttaa agatccgcgc
    11821 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
    11881 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
    11941 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
    12001 gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
    12061 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagta cctagatgtg
    12121 gcgcaacgat gccggcgaca agcaggagcg caccgacttc ttccgcatca agtgttttgg
    12181 ctctcaggcc gaggcccacg gcaagtattt gggcaagggg tcgctggtat tcgtgcaggg
    12241 caagattcgg aataccaagt acgagaagga cggccagacg gtctacggga ccgacttcat
    12301 tgccgataag gtggattatc tggacaccaa ggcaccaggc gggtcaaatc aggaataagg
    12361 gcacattgcc ccggcgtgag tcggggcaat cccgcaagga gggtgaatga atcggacgtt
    12421 tgaccggaag gcatacaggc aagaactgat cgacgcgggg ttttccgccg aggatgccga
    12481 aaccatcgca agccgcaccg tcatgcgtgc gccccgcgaa accttccagt ccgtcggctc
    12541 gatggtccag caagctacgg ccaagatcga gcgcgacagc gtgcaactgg ctccccctgc
    12601 cctgcccgcg ccatcggccg ccgtggagcg ttcgcgtcgt ctcgaacagg aggcggcagg
    12661 tttggcgaag tcgatgacca tcgacacgcg aggaactatg acgaccaaga agcgaaaaac
    12721 cgccggcgag gacctggcaa aacaggtcag cgaggccaag caggccgcgt tgctgaaaca
    12781 cacgaagcag cagatcaagg aaatgcagct ttccttgttc gatattgcgc cgtggccgga
    12841 cacgatgcga gcgatgccaa acgacacggc ccgctctgcc ctgttcacca cgcgcaacaa
    12901 gaaaatcccg cgcgaggcgc tgcaaaacaa ggtcattttc cacgtcaaca aggacgtgaa
    12961 gatcacctac accggcgtcg agctgcgggc cgacgatgac gaactggtgt ggcagcaggt
    13021 gttggagtac gcgaagcgca cccctatcgg cgagccgatc accttcacgt tctacgagct
    13081 ttgccaggac ctgggctggt cgatcaatgg ccggtattac acgaaggccg aggaatgcct
    13141 gtcgcgccta caggcgacgg cgatgggctt cacgtccgac cgcgttgggc acctggaatc
    13201 ggtgtcgctg ctgcaccgct tccgcgtcct ggaccgtggc aagaaaacgt cccgttgcca
    13261 ggtcctgatc gacgaggaaa tcgtcgtgct gtttgctggc gaccactaca cgaaattcat
    13321 atgggagaag taccgcaagc tgtcgccgac ggcccgacgg atgttcgact atttcagctc
    13381 gcaccgggag ccgtacccgc tcaagctgga aaccttccgc ctcatgtgcg gatcggattc
    13441 cacccgcgtg aagaagtggc gcgagcaggt cggcgaagcc tgcgaagagt tgcgaggcag
    13501 cggcctggtg gaacacgcct gggtcaatga tgacctggtg cattgcaaac gctagggcct
    13561 tgtggggtca gttccggctg ggggttcagc agccagcgct ttactggcat ttcaggaaca
    13621 agcgggcact gctcgacgca cttgcttcgc tcagtatcgc tcgggacgca cggcgcgctc
    13681 tacgaactgc cgataaacag aggattaaaa ttgacaattg tgattaaggc tcagattcga
    13741 cggcttggag cggccgacgt gcaggatttc cgcgagatcc gattgtcggc cctgaagaaa
    13801 gctccagaga tgttcgggtc cgtttacgag cacgaggaga aaaagcccat ggaggcgttc
    13861 gctgaacggt tgcgagatgc cgtggcattc ggcgcctaca tcgacggcga gatcattggg
    13921 ctgtcggtct tcaaacagga ggacggcccc aaggacgctc acaaggcgca tctgtccggc
    13981 gttttcgtgg agcccgaaca gcgaggccga ggggtcgccg gtatgctgct gcgggcgttg
    14041 ccggcgggtt tattgctcgt gatgatcgtc cgacagattc caacgggaat ctggtggatg
    14101 cgcatcttca tcctcggcgc acttaatatt tcgctattct ggagcttgtt gtttatttcg
    14161 gtctaccgcc tgccgggcgg ggtcgcggcg acggtaggcg ctgtgcagcc gctgatggtc
    14221 gtgttcatct ctgccgctct gctaggtagc ccgatacgat tgatggcggt cctgggggct
    14281 atttgcggaa ctgcgggcgt ggcgctgttg gtgttgacac caaacgcagc gctagatcct
    14341 gtcggcgtcg cagcgggcct ggcgggggcg gtttccatgg cgttcggaac cgtgctgacc
    14401 cgcaagtggc aacctcccgt gcctctgctc acctttaccg cctggcaact ggcggccgga
    14461 ggacttctgc tcgttccagt agctttagtg tttgatccgc caatcccgat gcctacagga
    14521 accaatgttc tcggcctggc gtggctcggc ctgatcggag cgggtttaac ctacttcctt
    14581 tggttccggg ggatctcgcg actcgaacct acagttgttt ccttactggg ctttctcagc
    14641 cccagatctg gggtcgatca gccggggatg catcaggccg acagtcggaa cttcgggtcc
    14701 ccgacctgta ccattcggtg agcaatggat aggggagttg atatcgtcaa cgttcacttc
    14761 taaagaaata gcgccactca gcttcctcag cggctttatc cagcgatttc ctattatgtc
    14821 ggcatagttc tcaagatcga cagcctgtca cggttaagcg agaaatgaat aagaaggctg
    14881 ataattcgga tctctgcgag ggagatgata tttgatcaca ggcagcaacg ctctgtcatc
    14941 gttacaatca acatgctacc ctccgcgaga tcatccgtgt ttcaaacccg gcagcttagt
    15001 tgccgttctt ccgaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct
    15061 cccgctgacg ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
    15121 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
    15181 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggtttttc
    15241 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
    15301 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggttc
    15361 cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt
    15421 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
    15481 aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg
    15541 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
    15601 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
    15661 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
    15721 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
    15781 agtcacgacg ttgtaaaacg acggccagtg aattaattcc cgatctagta acatagatga
    15841 caccgcgcgc gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta
    15901 aatgtataat tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta
    15961 catgttaatt attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag
    16021 accggcaaca ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatcggggaa
    16081 attcgagctc ggtaccccct cctgggcacg gtcgtacgcg atccgccgcg gttccacctc
    16141 accattgcgg gtatgccagt tgagttcgat gagttcccgt accagctcca actcgggcga
    16201 tgccacgcgc actccgttca gatctcggtg acgggcagga ccggacgggg cggtaccggc
    16261 aggctgaagt ccagctgcca gaaacccacg tcatgccagt tcccgtgctt gaagccggcc
    16321 gcccgcagca tgccgcgggg ggcatatccg agcgcctcgt gcatgcgcac gctcgggtcg
    16381 ttgggcagcc cgatgacagc gaccacgctc ttgaagccct gtgcctccag ggacttcagc
    16441 aggtgggtgt agagcgtgga gcccagtccc gtccgctggt ggcgggggga gacgtacacg
    16501 gtcgactcgg ccgtccagtc gtaggcgttg cgtgccttcc aggggcccgc gtaggcgatg
    16561 ccggcgacct cgccgtccac ctcggcgacg agccagggat agcgctcccg cagacggacg
    16621 aggtcgtccg tccactcctg cggttcctgc ggctcggtac ggaagttgac cgtgcttgtc
    16681 tcgatgtagt ggttgacgat ggtgcagacc gccggcatgt ccgcctcggt ggcacggcgg
    16741 atgtcggccg ggcgtcgttc tgggctcatg gatcctctag agtcccccgt gttctctcca
    16801 aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg gattgtgcgt
    16861 catcccttac gtcagtggag atatcacatc aatccacttg ctttgaagac gtggttggaa
    16921 cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg
    16981 cagaggcatc ttcaacgatg gcctttcctt tatcgcaatg atggcatttg taggagccac
    17041 cttccttttc cactatcttc acaataaagt gacagatagc tgggcaatgg aatccgagga
    17101 ggtttccgga tattaccctt tgttgaaaag tctcaattgc cctttggtct tctgagactg
    17161 tatctttgat atttttggag tagacaagtg tgtcgtgctc caccatgttg acgaagattt
    17221 tcttcttgtc attgagtcgt aagagactct gtatgaactg ttcgccagtc tttacggcga
    17281 gttctgttag gtcctctatt tgaatctttg actccatggc ctttgattca gtgggaacta
    17341 cctttttaga gactccaatc tctattactt gccttggttt gtgaagcaag ccttgaatcg
    17401 tccatactgg aatagtactt ctgatcttga gaaatatatc tttctctgtg ttcttgatgc
    17461 agttagtcct gaatcttttg actgcatctt taaccttctt gggaaggtat ttgatttcct
    17521 ggagattatt gctcgggtag atcgtcttga tgagacctgc tgcgtaagcc tctctaacca
    17581 tctgtgggtt agcattcttt ctgaaattga aaaggctaat ctggggacct gcaggggcaa
    17641 taccgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctttcga
    17701 cgagtcagta ataaacggcg tcaaagtggt tgcagccggc acacacgagt cgtgtttatc
    17761 aactcaaagc acaaatactt ttcctcaacc taaaaataag gcaattagcc aaaaacaact
    17821 ttgcgtgtaa acaacgctca atacacgtgt cattttatta ttagctattg cttcaccgcc
    17881 ttagctttct cgtgacctag tcgtcctcgt cttttcttct tcttcttcta taaaacaata
    17941 cccaaagagc tcttcttctt cacaattcag atttcaattt ctcaaaatct taaaaacttt
    18001 ctctcaattc tctctaccgt gatcaaggta aatttctgtg ttccttattc tctcaaaatc
    18061 ttcgattttg ttttcgttcg atcccaattt cgtatatgtt ctttggttta gattctgtta
    18121 atcttagatc gaagacgatt ttctgggttt gatcgttaga tatcatctta attctcgatt
    18181 agggtttcat agatatcatc cgatttgttc aaataatttg agttttgtcg aataattact
    18241 cttcgatttg tgatttctat ctagatctgg tgttagtttc tagtttgtgc gatcgaattt
    18301 gtagattaat ctgagttttt ctgattaaca gggtac
//
