LOCUS       sequence-202609-       18890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Cas9 2xplex gRNA targeting Medtr2g044580 (ERDJ2). Also 
            expresses Cas9 from Gmubi promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18890)
  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 18890)
  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: pSC13
FEATURES             Location/Qualifiers
     source          1..18890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(145..516)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            549..658
                     /note="incP origin of transfer"
     primer_bind     881..900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1134..1151
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1339..1989)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    2454..2478
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    3686..3710
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3732..3779
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             3786..3806
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3831..5267,5457..8120)
                     /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          5268..5456
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             8121..8168
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8179..8431
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    8476..8496
                     /gene="mutant version of attB"
                     /label=attB5
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        8851..8926
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(8938..8962)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        9440..9515
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(9527..9551)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(9570..9594)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      9632..10339
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     11177..11197
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             11274..11825
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     terminator      11851..12103
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(12116..12133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12116..12132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12125..12147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(12272..12291)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12482..12503
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    12750..12774
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             complement(14370..15518)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(15817..16611)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     mobile_element  16803..17570
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     rep_origin      join(18263..18890,1..8)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tgagcgtcgg aaaggcgctc ggtcttgcct tgctcgtcgg tgatgtactt caccagctcc
       61 gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac gcgcaccccc
      121 cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca cgcccatcat
      181 gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga ggatcgtggc
      241 atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca ggccgcccag
      301 gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt ccacgacgcc
      361 cgtgattttg tagccctggc cgacggccag caggtaggcc gacaggctca tgccggccgc
      421 cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct tgataggtgg
      481 gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat ctgttacgcc
      541 ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca ggtgcgaata
      601 agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta tcctgcccgg
      661 ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa atcctgtata
      721 tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga agcagggtta
      781 tgcagcggaa aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
      841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1021 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1081 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1141 cagtgagcga ggaagcggaa gagcgccaga aggccgccag agaggccgag cgcggccgtg
     1201 aggcttggac gctagggcag ggcatgaaaa agcccgtagc gggctgctac gggcgtctga
     1261 cgcggtggaa agggggaggg gatgttgtct acatggctct gctgtagtga gtgggttgcg
     1321 ctccggcagc ggtcctgatc aatcgtcacc ctttctcggt ccttcaacgt tcctgacaac
     1381 gagcctcctt ttcgccaatc catcgacaat caccgcgagt ccctgctcga acgctgcgtc
     1441 cggaccggct tcgtcgaagg cgtctatcgc ggcccgcaac agcggcgaga gcggagcctg
     1501 ttcaacggtg ccgccgcgct cgccggcatc gctgtcgccg gcctgctcct caagcacggc
     1561 cccaacagtg aagtagctga ttgtcatcag cgcattgacg gcgtccccgg ccgaaaaacc
     1621 cgcctcgcag aggaagcgaa gctgcgcgtc ggccgtttcc atctgcggtg cgcccggtcg
     1681 cgtgccggca tggatgcgcg cgccatcgcg gtaggcgagc agcgcctgcc tgaagctgcg
     1741 ggcattcccg atcagaaatg agcgccagtc gtcgtcggct ctcggcaccg aatgcgtatg
     1801 attctccgcc agcatggctt cggccagtgc gtcgagcagc gcccgcttgt tcctgaagtg
     1861 ccagtaaagc gccggctgct gaacccccaa ccgttccgcc agtttgcgtg tcgtcagacc
     1921 gtctacgccg acctcgttca acaggtccag ggcggcacgg atcactgtat tcggctgcaa
     1981 ctttgtcatg cttgacactt tatcactgat aaacataata tgtccaccaa cttatcagtg
     2041 ataaagaatc cgcgcgttca atcggaccag cggaggctgg tccggaggcc agacgtgaaa
     2101 cccaacatac ccctgatcgt aattctgagc actgtcgcgc tcgacgctgt cggcatcggc
     2161 ctgattatgc cggtgctgcc gggcctcctg cgcgatctgg ttcactcgaa cgacgtcacc
     2221 gcccactatg gcattctgct ggcgctgtat gcgttggtgc aatttgcctg cgcacctgtg
     2281 ctgggcgcgc tgtcggatcg tttcgggcgg cggccaatct tgctcgtctc gctggccggc
     2341 gccagatctg gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
     2401 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
     2461 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
     2521 tcatgagcgg agaattaagg gagtcacgtt atgacccccg ccgatgacgc gggacaagcc
     2581 gttttacgtt tggaactgac agaaccgcaa cgttgaagga gccactcagc cgcgggtttc
     2641 tggagtttaa tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc
     2701 taaggtcact atcagctaga cggacgcggc cgcgggccca atataacaac gacgtcgtaa
     2761 cagataaagc gaagcttgaa ggtgcatgtg actccgtcaa gattacgaaa ccgccaacta
     2821 ccacgcaaat tgcaattctc aatttcctag aaggactctc cgaaaatgca tccaatacca
     2881 aatattaccc gtgtcatagg caccaagtga caccatacat gaacacgcgt cacaatatga
     2941 ctggagaagg gttccacacc ttatgctata aaacgcccca cacccctcct ccttccttcg
     3001 cagttcaatt ccaatatatt ccattctctc tgtgtatttc cctacctctc ccttcaaggt
     3061 tagtcgattt cttctgtttt tcttcttcgt tctttccatg aattgtgtat gttctttgat
     3121 caatacgatg ttgatttgat tgtgttttgt ttggtttcat cgatcttcaa ttttcataat
     3181 cagattcagc ttttattatc tttacaacaa cgtccttaat ttgatgattc tttaatcgta
     3241 gatttgctct aattagagct ttttcatgtc agatcccttt acaacaagcc ttaattgttg
     3301 attcattaat cgtagattag ggcttttttc attgattact tcagatccgt taaacgtaac
     3361 catagatcag ggctttttca tgaattactt cagatccgtt aaacaacagc cttatttttt
     3421 atacttctgt ggtttttcaa gaaattgttc agatccgttg acaaaaagcc ttattcgttg
     3481 attctatatc gtttttcgag agatattgct cagatctgtt agcaactgcc ttgtttgttg
     3541 attctattgc cgtggattag ggtttttttt cacgagattg cttcagatcc gtacttaaga
     3601 ttacgtaatg gattttgatt ctgatttatc tgtgattgtt gactcgacag gtaccttcaa
     3661 acctcgaggc gcgccaagct atcaaacaag tttgtacaaa aaagcaggct ccgaattcgc
     3721 ccttcaccat ggattacaag gatgatgatg ataaggatta caaggatgat gatgataaga
     3781 tggctccaaa gaagaagaga aaggttggaa tccacggagt tccagctgct gataagaagt
     3841 actctatcgg acttgacatc ggaaccaact ctgttggatg ggctgttatc accgatgagt
     3901 acaaggttcc atctaagaag ttcaaggttc ttggaaacac cgatagacac tctatcaaga
     3961 agaaccttat cggtgctctt cttttcgatt ctggagagac cgctgaggct accagattga
     4021 agagaaccgc tagaagaaga tacaccagaa gaaagaacag aatctgctac cttcaggaaa
     4081 tcttctctaa cgagatggct aaggttgatg attctttctt ccacagactt gaggagtctt
     4141 tccttgttga ggaggataag aagcacgaga gacacccaat cttcggaaac atcgttgatg
     4201 aggttgctta ccacgagaag tacccaacca tctaccacct tagaaagaag ttggttgatt
     4261 ctaccgataa ggctgatctt agacttatct accttgctct tgctcacatg atcaagttca
     4321 gaggacactt ccttatcgag ggagacctta acccagataa ctctgatgtt gataagttgt
     4381 tcatccagct tgttcagacc tacaaccagc ttttcgagga gaacccaatc aacgcttctg
     4441 gagttgatgc taaggctatc ctttctgcta gactttctaa gtctcgtaga cttgagaacc
     4501 ttatcgctca gcttccagga gagaagaaga acggactttt cggaaacctt atcgctcttt
     4561 ctcttggact taccccaaac ttcaagtcta acttcgatct tgctgaggat gctaagttgc
     4621 agctttctaa ggatacctac gatgatgatc ttgataacct tcttgctcag atcggagatc
     4681 agtacgctga tcttttcctt gctgctaaga acctttctga tgctatcctt ctttctgaca
     4741 tccttagagt taacaccgag atcaccaagg ctccactttc tgcttctatg atcaagagat
     4801 acgatgagca ccaccaggat cttacccttt tgaaggctct tgttagacag cagcttccag
     4861 agaagtacaa ggaaatcttc ttcgatcagt ctaagaacgg atacgctgga tacatcgatg
     4921 gaggagcttc tcaggaggag ttctacaagt tcatcaagcc aatccttgag aagatggatg
     4981 gaaccgagga gcttcttgtt aagttgaaca gagaggatct tcttagaaag cagagaacct
     5041 tcgataacgg atctatccca caccagatcc accttggaga gcttcacgct atccttcgta
     5101 gacaggagga tttctaccca ttcttgaagg ataacagaga gaagatcgag aagatcctta
     5161 ccttcagaat cccatactac gttggaccac ttgctagagg aaactctcgt ttcgcttgga
     5221 tgaccagaaa gtctgaggag accatcaccc cttggaactt cgaggaggta agtttctgct
     5281 tctacctttg atatatatat aataattatc attaattagt agtaatataa tatttcaaat
     5341 atttttttca aaataaaaga atgtagtata tagcaattgc ttttctgtag tttataagtg
     5401 tgtatatttt aatttataac ttttctaata tatgaccaaa atttgttgat gtgcaggttg
     5461 ttgataaggg agcttctgct cagtctttca tcgagagaat gaccaacttc gataagaacc
     5521 ttccaaacga gaaggttctt ccaaagcact ctcttcttta cgagtacttc accgtttaca
     5581 acgagcttac caaggttaag tacgttaccg agggaatgag aaagccagct ttcctttctg
     5641 gagagcagaa gaaggctatc gttgatcttc ttttcaagac caacagaaag gttaccgtta
     5701 agcagttgaa ggaggattac ttcaagaaga tcgagtgctt cgattctgtt gaaatctctg
     5761 gagttgagga tagattcaac gcttctcttg gaacctacca cgatcttttg aagatcatca
     5821 aggataagga tttccttgat aacgaggaga acgaggacat ccttgaggac atcgttctta
     5881 cccttaccct tttcgaggat agagagatga tcgaggagag actcaagacc tacgctcacc
     5941 ttttcgatga taaggttatg aagcagttga agagaagaag atacaccgga tggggtagac
     6001 tttctcgtaa gttgatcaac ggaatcagag ataagcagtc tggaaagacc atccttgatt
     6061 tcttgaagtc tgatggattc gctaacagaa acttcatgca gcttatccac gatgattctc
     6121 ttaccttcaa ggaggacatc cagaaggctc aggtttctgg acagggagat tctcttcacg
     6181 agcacatcgc taaccttgct ggatctccag ctatcaagaa gggaatcctt cagaccgtta
     6241 aggttgttga tgagcttgtt aaggttatgg gtagacacaa gccagagaac atcgttatcg
     6301 agatggctag agagaaccag accacccaga agggacagaa gaactctcgt gagagaatga
     6361 agagaatcga ggagggaatc aaggagcttg gatctcaaat cttgaaggag cacccagttg
     6421 agaacaccca gcttcagaac gagaagttgt acctttacta ccttcagaac ggaagagata
     6481 tgtacgttga tcaggagctt gacatcaaca gactttctga ttacgatgtt gatcacatcg
     6541 ttccacagtc tttcttgaag gatgattcta tcgataacaa ggttcttacc cgttctgata
     6601 agaacagagg aaagtctgat aacgttccat ctgaggaggt tgttaagaag atgaagaact
     6661 actggagaca gcttcttaac gctaagttga tcacccagag aaagttcgat aaccttacca
     6721 aggctgagag aggaggactt tctgagcttg ataaggctgg attcatcaag agacagcttg
     6781 ttgagaccag acagatcacc aagcacgttg ctcagatcct tgattctcgt atgaacacca
     6841 agtacgatga gaacgataag ttgatcagag aggttaaggt tatcaccttg aagtctaagt
     6901 tggtttctga tttcagaaag gatttccagt tctacaaggt tagagagatc aacaactacc
     6961 accacgctca cgatgcttac cttaacgctg ttgttggaac cgctcttatc aagaagtacc
     7021 caaagttgga gtctgagttc gtttacggag attacaaggt ttacgatgtt agaaagatga
     7081 tcgctaagtc tgagcaggag atcggaaagg ctaccgctaa gtacttcttc tactctaaca
     7141 tcatgaactt cttcaagacc gagatcaccc ttgctaacgg agagatcaga aagagaccac
     7201 ttatcgagac caacggagag accggagaga tcgtttggga taagggaaga gatttcgcta
     7261 ccgttagaaa ggttctttct atgccacagg ttaacatcgt taagaaaacc gaggttcaga
     7321 ccggaggatt ctctaaggag tctatccttc caaagagaaa ctctgataag ttgatcgcta
     7381 gaaagaagga ttgggaccca aagaagtacg gaggattcga ttctccaacc gttgcttact
     7441 ctgttcttgt tgttgctaag gttgagaagg gaaagtctaa gaagttgaag tctgttaagg
     7501 agcttcttgg aatcaccatc atggagcgtt cttctttcga gaagaaccca atcgatttcc
     7561 ttgaggctaa gggatacaag gaggttaaga aggatcttat catcaagttg ccaaagtact
     7621 ctcttttcga gcttgagaac ggaagaaaga gaatgcttgc ttctgctgga gagcttcaga
     7681 agggaaacga gcttgctctt ccatctaagt acgttaactt cctttacctt gcttctcact
     7741 acgagaagtt gaagggatct ccagaggata acgagcagaa gcagcttttc gttgagcagc
     7801 acaagcacta ccttgatgag atcatcgagc aaatctctga gttctctaag agagttatcc
     7861 ttgctgatgc taaccttgat aaggttcttt ctgcttacaa caagcacaga gataagccaa
     7921 tcagagagca ggctgagaac atcatccacc ttttcaccct taccaacctt ggtgctccag
     7981 ctgctttcaa gtacttcgat accaccatcg atagaaaaag atacacctct accaaggagg
     8041 ttcttgatgc tacccttatc caccagtcta tcaccggact ttacgagacc agaatcgatc
     8101 tttctcagct tggaggagat aagagaccag ctgctaccaa gaaggctgga caggctaaga
     8161 agaagaagtg agacgtccga tcgttcaaac atttggcaat aaagtttctt aagattgaat
     8221 cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt taagcatgta
     8281 ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
     8341 caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta ggataaatta
     8401 tcgcgcgcgg tgtcatctat gttactagat cgggaattga tcccccctcg acagcttccg
     8461 gaaagggcga attcgcaact ttgtatacaa aagttgccga gctcgcggcc gcatgttgtt
     8521 gttaccagaa agtaaataaa tgttcaatct ctgatgttct caagtaagtg agttttattg
     8581 ggaataatat taacttatgt tcttcttgca tttgatttct ttgccgctct cttcttctat
     8641 cttaaatctg tgtatactat ttcactattg ggctttttat tagtctataa tgggactcaa
     8701 aataaggctt tggcccacat caaaaagata agtcacaaat caaaactaaa ttcagagtct
     8761 tttctcccac atcggtcact gtactcattt tgtgtttgtt tatatattac acgaaccgat
     8821 ctttggtacg gtactgaagg ttagcgacga gttttagagc tagaaatagc aagttaaaat
     8881 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctagacc
     8941 cagctttctt gtacaaagtt ggcattacca tggcttcgtt gaacaacgga aactcgactt
     9001 gccttccgca caatacatca tttcttctta gctttttttc ttcttcttcg ttcatacagt
     9061 ttttttttgt ttatcagctt acattttctt gaaccgtagc tttcgttttc ttctttttaa
     9121 ctttccattc ggagtttttg tatcttgttt catagtttgt cccaggatta gaatgattag
     9181 gcatcgaacc ttcaagaatt tgattgaata aaacatcttc attcttaaga tatgaagata
     9241 atcttcaaaa ggcccctggg aatctgaaag aagagaagca ggcccattta tatgggaaag
     9301 aacaatagta tttcttatat aggcccattt aagttgaaaa caatcttcaa aagtcccaca
     9361 tcgcttagat aagaaaacga agctgagttt atatacagct agagtcgaag tagtgattga
     9421 atattgatgg ggcatctggg ttttagagct agaaatagca agttaaaata aggctagtcc
     9481 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttctagaccc agctttcttg
     9541 tacaaagttg gcattactgc aggtcgacga cccagctttc ttgtacaaag tggttcgata
     9601 attcttaatt aactagttct aatctagagt cctgctttaa tgagatatgc gagacgccta
     9661 tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt gtaaaaaacc tgagcatgtg
     9721 tagctcagat ccttaccgcc ggtttcggtt cattctaatg aatatatcac ccgttactat
     9781 cgtattttta tgaataatat tctccgttca atttactgat tgtaccctac tacttatatg
     9841 tacaatatta aaatgaaaac aatatattgt gctgaatagg tttatagcga catctatgat
     9901 agagcgccac aataacaaac aattgcgttt tattattaca aatccaattt taaaaaaagc
     9961 ggcagaaccg gtcaaaccta aaagactgat tacataaatc ttattcaaat ttcaaaagtg
    10021 ccccaggggc tagtatctac gacacaccga gcggcgaact aataacgctc actgaaggga
    10081 actccggttc cccgccggcg cgcatgggtg agattccttg aagttgagta ttggccgtcc
    10141 gctctaccga aagttacggg caccattcaa cccggtccag cacggcggcc gggtaaccga
    10201 cttgctgccc cgagaattat gcagcatttt tttggtgtat gtgggcccca aatgaagtgc
    10261 aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt ccagggcgaa ttttgcgaca
    10321 acatgtcgag gctcagcagg acctgcaggc atgcaagcta gtgaagaatc gatttgctac
    10381 gtattgcccc tgcaggtccc cagattagcc ttttcaattt cagaaagaat gctaacccac
    10441 agatggttag agaggcttac gcagcaggtc tcatcaagac gatctacccg agcaataatc
    10501 tccaggaaat caaatacctt cccaagaagg ttaaagatgc agtcaaaaga ttcaggacta
    10561 actgcatcaa gaacacagag aaagatatat ttctcaagat cagaagtact attccagtat
    10621 ggacgattca aggcttgctt cacaaaccaa ggcaagtaat agagattgga gtctctaaaa
    10681 aggtagttcc cactgaatca aaggccatgg agtcaaagat tcaaatagag gacctaacag
    10741 aactcgccgt aaagactggc gaacagttca tacagagtct cttacgactc aatgacaaga
    10801 agaaaatctt cgtcaacatg gtggagcacg acacacttgt ctactccaaa aatatcaaag
    10861 atacagtctc agaagaccaa agggcaattg agacttttca acaaagggta atatccggaa
    10921 acctcctcgg attccattgc ccagctatct gtcactttat tgtgaagata gtggaaaagg
    10981 aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt gaagatgcct
    11041 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
    11101 acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg
    11161 atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc
    11221 atataaggaa gttcatttca tttggagaga acacggggga ctctagagga tccatgagcc
    11281 cagaacgacg cccggccgac atccgccgtg ccaccgaggc ggacatgccg gcggtctgca
    11341 ccatcgtcaa ccactacatc gagacaagca cggtcaactt ccgtaccgag ccgcaggaac
    11401 cgcaggagtg gacggacgac ctcgtccgtc tgcgggagcg ctatccctgg ctcgtcgccg
    11461 aggtggacgg cgaggtcgcc ggcatcgcct acgcgggccc ctggaaggca cgcaacgcct
    11521 acgactggac ggccgagtcg accgtgtacg tctccccccg ccaccagcgg acgggactgg
    11581 gctccacgct ctacacccac ctgctgaagt ccctggaggc acagggcttc aagagcgtgg
    11641 tcgctgtcat cgggctgccc aacgacccga gcgtgcgcat gcacgaggcg ctcggatatg
    11701 ccccccgcgg catgctgcgg gcggccggct tcaagcacgg gaactggcat gacgtgggtt
    11761 tctggcagct ggacttcagc ctgccggtac cgccccgtcc ggtcctgccc gtcaccgaga
    11821 tctgaggggg taccgagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc
    11881 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
    11941 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
    12001 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
    12061 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt aattcactgg
    12121 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
    12181 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
    12241 cccaacagtt gcgcagcctg aatggcgccc gctcctttcg ctttcttccc ttcctttctc
    12301 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
    12361 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
    12421 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    12481 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
    12541 ttataaggga ttttgccgat ttcggaacca ccatcaaaca ggattttcgc ctgctggggc
    12601 aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc
    12661 tgttgcccgt ctcactggtg aaaagaaaaa ccaccccagt acattaaaaa cgtccgcaat
    12721 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
    12781 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
    12841 tcagtccggg acggcgtcag cgggagagcc gttgtaaggc ggcagacttt gctcatgtta
    12901 ccgatgctat tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc
    12961 ggagggtagc atgttgattg taacgatgac agagcgttgc tgcctgtgat caaatatcat
    13021 ctccctcgca gagatccgaa ttatcagcct tcttattcat ttctcgctta accgtgacag
    13081 gctgtcgatc ttgagaacta tgccgacata ataggaaatc gctggataaa gccgctgagg
    13141 aagctgagtg gcgctatttc tttagaagtg aacgttgacg atatcaactc ccctatccat
    13201 tgctcaccga atggtacagg tcggggaccc gaagttccga ctgtcggcct gatgcatccc
    13261 cggctgatcg accccagatc tggggctgag aaagcccagt aaggaaacaa ctgtaggttc
    13321 gagtcgcgag atcccccgga accaaaggaa gtaggttaaa cccgctccga tcaggccgag
    13381 ccacgccagg ccgagaacat tggttcctgt aggcatcggg attggcggat caaacactaa
    13441 agctactgga acgagcagaa gtcctccggc cgccagttgc caggcggtaa aggtgagcag
    13501 aggcacggga ggttgccact tgcgggtcag cacggttccg aacgccatgg aaaccgcccc
    13561 cgccaggccc gctgcgacgc cgacaggatc tagcgctgcg tttggtgtca acaccaacag
    13621 cgccacgccc gcagttccgc aaatagcccc caggaccgcc atcaatcgta tcgggctacc
    13681 tagcagagcg gcagagatga acacgaccat cagcggctgc acagcgccta ccgtcgccgc
    13741 gaccccgccc ggcaggcggt agaccgaaat aaacaacaag ctccagaata gcgaaatatt
    13801 aagtgcgccg aggatgaaga tgcgcatcca ccagattccc gttggaatct gtcggacgat
    13861 catcacgagc aataaacccg ccggcaacgc ccgcagcagc ataccggcga cccctcggcc
    13921 tcgctgttcg ggctccacga aaacgccgga cagatgcgcc ttgtgagcgt ccttggggcc
    13981 gtcctcctgt ttgaagaccg acagcccaat gatctcgccg tcgatgtagg cgccgaatgc
    14041 cacggcatct cgcaaccgtt cagcgaacgc ctccatgggc tttttctcct cgtgctcgta
    14101 aacggacccg aacatctctg gagctttctt cagggccgac aatcggatct cgcggaaatc
    14161 ctgcacgtcg gccgctccaa gccgtcgaat ctgagcctta atcacaattg tcaattttaa
    14221 tcctctgttt atcggcagtt cgtagagcgc gccgtgcgtc ccgagcgata ctgagcgaag
    14281 caagtgcgtc gagcagtgcc cgcttgttcc tgaaatgcca gtaaagcgct ggctgctgaa
    14341 cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag gtcatcattg
    14401 acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc gccgacctgc
    14461 tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc
    14521 ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc
    14581 gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg
    14641 acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg gtccaggacg
    14701 cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc
    14761 atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata ccggccattg
    14821 atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata
    14881 ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc gtcggcccgc
    14941 agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat gaccttgttt
    15001 tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga gcgggccgtg
    15061 tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc
    15121 atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc
    15181 tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg
    15241 tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc
    15301 acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc
    15361 ttggccgtag cttgctggac catcgagccg acggactgga aggtttcgcg gggcgcacgc
    15421 atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt
    15481 tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg cgggattgcc
    15541 ccgactcacg ccggggcaat gtgcccttat tcctgatttg acccgcctgg tgccttggtg
    15601 tccagataat ccaccttatc ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc
    15661 tcgtacttgg tattccgaat cttgccctgc acgaatacca gcgacccctt gcccaaatac
    15721 ttgccgtggg cctcggcctg agagccaaaa cacttgatgc ggaagaagtc ggtgcgctcc
    15781 tgcttgtcgc cggcatcgtt gcgccacatc taggtactaa aacaattcat ccagtaaaat
    15841 ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac
    15901 atactgttct tccccgatat cctccctgat cgaccggacg cagaaggcaa tgtcatacca
    15961 cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac
    16021 aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc
    16081 cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt
    16141 ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct
    16201 gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca
    16261 ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca
    16321 aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg
    16381 caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc
    16441 cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt
    16501 ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg
    16561 gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tttattattt
    16621 ccttcctctt ttctacagta tttaaagata ccccaagaag ctaattataa caagacgaac
    16681 tccaattcac tgttccttgc attctaaaac cttaaatacc agaaaacagc tttttcaaag
    16741 ttgttttcaa agttggcgta taacatagta tcgacggagc cgattttgaa accacaatta
    16801 tgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg
    16861 gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc
    16921 aaaagcaccg ccggacatca gcgctatctc tgctctcact gccgtaaaac atggcaactg
    16981 cagttcactt acaccgcttc tcaacccggt acgcaccaga aaatcattga tatggccatg
    17041 aatggcgttg gatgccgggc aacagcccgc attatgggcg ttggcctcaa cacgatttta
    17101 cgtcacttaa aaaactcagg ccgcagtcgg taacctcgcg catacagccg ggcagtgacg
    17161 tcatcgtctg cgcggaaatg gacgaacagt ggggctatgt cggggctaaa tcgcgccagc
    17221 gctggctgtt ttacgcgtat gacagtctcc ggaagacggt tgttgcgcac gtattcggtg
    17281 aacgcactat ggcgacgctg gggcgtctta tgagcctgct gtcacccttt gacgtggtga
    17341 tatggatgac ggatggctgg ccgctgtatg aatcccgcct gaagggaaag ctgcacgtaa
    17401 tcagcaagcg atatacgcag cgaattgagc ggcataacct gaatctgagg cagcacctgg
    17461 cacggctggg acggaagtcg ctgtcgttct caaaatcggt ggagctgcat gacaaagtca
    17521 tcgggcatta tctgaacata aaacactatc aataagttgg agtcattacc caattatgat
    17581 agaatttaca agctataagg ttattgtcct gggtttcaag cattagtcca tgcaagtttt
    17641 tatgctttgc ccattctata gatatattga taagcgcgct gcctatgcct tgccccctga
    17701 aatccttaca tacggcgata tcttctatat aaaagatata ttatcttatc agtattgtca
    17761 atatattcaa ggcaatctgc ctcctcatcc tcttcatcct cttcgtcttg gtagcttttt
    17821 aaatatggcg cttcatagag taattctgta aaggtccaat tctcgttttc atacctcggt
    17881 ataatcttac ctatcacctc aaatggttcg ctgggtttat cgcacccccg aacacgagca
    17941 cggcacccgc gaccactatg ccaagaatgc ccaaggtaaa aattgccggc cccgccatga
    18001 agtccgtgaa tgccccgacg gccgaagtga agggcaggcc gccacccagg ccgccgccct
    18061 cactgcccgg cacctggtcg ctgaatgtcg atgccagcac ctgcggcacg tcaatgcttc
    18121 cgggcgtcgc gctcgggctg atcgcccatc ccgttactgc cccgatcccg gcaatggcaa
    18181 ggactgccag cgctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct
    18241 ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc
    18301 ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat
    18361 attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc
    18421 ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct
    18481 catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc
    18541 gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt
    18601 gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc
    18661 gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct
    18721 caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca
    18781 caacgccggc ggccggccgc ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt
    18841 tgcagggcca tagacggccg ccagcccagc ggcgagggca accagcccgg
//
