LOCUS       sequence-182721-       15703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for targeted deletion of 58kb region in medicago 
            truncatula (Csy4 array of 6 gRNAs under the control of CmYLCV 
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15703)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 15703)
  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"; journalName: "Plant Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pSC3
FEATURES             Location/Qualifiers
     source          1..15703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(291..1085)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    1510..1534
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    1612..1786
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1793..2344)
                     /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(2389..3065)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    3256..3277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3292..3297,3298..3315,3316..3322)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3330..3346
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3335..3357
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3354..3370
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3354..3370
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3900..4245
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             4837..4893
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4894..8997
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4894..8997
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9010..9030
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9041..9289
                     /label=HSP terminator
                     /note="efficient transcription terminator from the 
                     Arabidopsis thaliana heat shock protein 18.2 gene (Nagaya 
                     et al., 2010)"
     protein_bind    9786..9805
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        9826..9901
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    9902..9921
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        9942..10017
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10018..10037
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        10058..10133
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10134..10153
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        10174..10249
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10250..10269
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        10290..10365
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10366..10385
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        10406..10481
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    10482..10501
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     polyA_signal    10521..10695
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(10714..10731)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10714..10730)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10723..10745)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    10933..10957
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             12258..12887
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             13316..14389
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      14455..14649
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(14885..14907)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    14993..15133
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     15048..15067
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(15148..15165)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(15319..15703,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15399..15418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
       61 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta ctaaaacaat
      301 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
      361 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag
      421 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
      481 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
      541 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
      601 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
      661 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
      721 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
      781 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
      841 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
      901 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
      961 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
     1021 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
     1081 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
     1141 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
     1201 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
     1261 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc
     1321 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat
     1381 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac
     1441 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
     1501 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
     1561 cacattgcgg acgtttttaa tgtactgaat taacgccgaa ttaattcggg ggatctggat
     1621 tttagtactg gattttggtt ttaggaatta gaaattttat tgatagaagt attttacaaa
     1681 tacaaataca tactaagggt ttcttatatg ctcaacacat gagcgaaacc ctataggaac
     1741 cctaattccc ttatctggga actactcaca cattattatg gagaaactcg agtcaaatct
     1801 cggtgacggg caggaccgga cggggcggta ccggcaggct gaagtccagc tgccagaaac
     1861 ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat
     1921 atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca
     1981 cgctcttgaa gccctgtgcc tccagggact tcagcagatg ggtgtagagc gtggagccca
     2041 gtcccgtccg ctggtggcgg gggctgacgt acacggtcga ctcggccgtc cagtcgtagg
     2101 cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc gacctcgccg tccacctcgg
     2161 cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt
     2221 cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc
     2281 agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc
     2341 tcatggtaga ctcgagagag atagatttgt agagagagac tggtgatttc agcgtgtcct
     2401 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
     2461 tgcgtcatcc cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt
     2521 tggaacgtct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
     2581 gtcggcagag gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt
     2641 gccaccttcc ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc
     2701 gaggaggttt cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga
     2761 gactgtatct ttgatattct tggagtagac gagagtgtcg tgctccacca tgttcacatc
     2821 aatccacttg ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg
     2881 gtgggggtcc atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt
     2941 tatcgcaatg atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt
     3001 gacagatagc tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag
     3061 tctcaatagc cctttggtct tctgagactg tatctttgat attcttggag tagacgagag
     3121 tgtcgtgctc caccatgttg gcaagctgct ctagccaata cgcaaaccgc ctctccccgc
     3181 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3241 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3301 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3361 cagctatgac atgattacga attcgagctc ggtacccggg gatcggcgcg ccagatttgc
     3421 cttttcaatt tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcaggt
     3481 atcatcaaga cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag
     3541 gttaaagatg cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata
     3601 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca
     3661 aggcaagtaa tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg
     3721 gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc
     3781 atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac
     3841 gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt
     3901 gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc
     3961 tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc
     4021 gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc
     4081 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
     4141 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
     4201 gacccttcct ctatataagg aagttcattt catttggaga gaacacgggg gactcctgca
     4261 ggaaaaatgg atcattatct tgatattaga cttagacctg atccagaatt tccaccagct
     4321 caacttatgt ctgttctttt tggaaaactt catcaagctc ttgttgctca aggaggagat
     4381 agaattggag tttcttttcc tgatcttgat gaatcaagat caagacttgg agaaagactt
     4441 agaattcatg cttctgctga tgatcttaga gctttgcttg ctagaccttg gcttgaagga
     4501 cttagagatc atcttcaatt tggagaacca gctgttgttc cacatccaac tccttataga
     4561 caagtttcaa gagttcaagc taaatctaat ccagaaagac ttagaagaag acttatgaga
     4621 agacatgatc tttctgaaga agaagctaga aaaagaattc ctgatactgt tgctagagct
     4681 ttggatttgc cttttgttac acttagatca caatctactg gacaacattt tagacttttt
     4741 attagacatg gaccacttca agttactgct gaagaaggag gatttacttg ttatggactt
     4801 tctaagggag gttttgttcc ttggtttgga tctggagcta ctaatttttc tcttcttaag
     4861 caagctggag atgttgaaga aaatcctgga cccatggata agaagtactc tatcggactc
     4921 gatatcggaa ctaactctgt gggatgggct gtgatcaccg atgagtacaa ggtgccatct
     4981 aagaagttca aggttctcgg aaacaccgat aggcactcta tcaagaaaaa ccttatcggt
     5041 gctctcctct tcgattctgg tgaaactgct gaggctacca gactcaagag aaccgctaga
     5101 agaaggtaca ccagaagaaa gaacaggatc tgctacctcc aagagatctt ctctaacgag
     5161 atggctaaag tggatgattc attcttccac aggctcgaag agtcattcct cgtggaagaa
     5221 gataagaagc acgagaggca ccctatcttc ggaaacatcg ttgatgaggt ggcataccac
     5281 gagaagtacc ctactatcta ccacctcaga aagaagctcg ttgattctac tgataaggct
     5341 gatctcaggc tcatctacct cgctctcgct cacatgatca agttcagagg acacttcctc
     5401 atcgagggtg atctcaaccc tgataactct gatgtggata agttgttcat ccagctcgtg
     5461 cagacctaca accagctttt cgaagagaac cctatcaacg cttcaggtgt ggatgctaag
     5521 gctatcctct ctgctaggct ctctaagtca agaaggcttg agaacctcat tgctcagctc
     5581 cctggtgaga agaagaacgg acttttcgga aacttgatcg ctctctctct cggactcacc
     5641 cctaacttca agtctaactt cgatctcgct gaggatgcaa agctccagct ctcaaaggat
     5701 acctacgatg atgatctcga taacctcctc gctcagatcg gagatcagta cgctgatttg
     5761 ttcctcgctg ctaagaacct ctctgatgct atcctcctca gtgatatcct cagagtgaac
     5821 accgagatca ccaaggctcc actctcagct tctatgatca agagatacga tgagcaccac
     5881 caggatctca cacttctcaa ggctcttgtt agacagcagc tcccagagaa gtacaaagag
     5941 attttcttcg atcagtctaa gaacggatac gctggttaca tcgatggtgg tgcatctcaa
     6001 gaagagttct acaagttcat caagcctatc ctcgagaaga tggatggaac cgaggaactc
     6061 ctcgtgaagc tcaatagaga ggatcttctc agaaagcaga ggaccttcga taacggatct
     6121 atccctcatc agatccacct cggagagttg cacgctatcc ttagaaggca agaggatttc
     6181 tacccattcc tcaaggataa cagggaaaag attgagaaga ttctcacctt cagaatccct
     6241 tactacgtgg gacctctcgc tagaggaaac tcaagattcg cttggatgac cagaaagtct
     6301 gaggaaacca tcaccccttg gaacttcgaa gaggtggtgg ataagggtgc tagtgctcag
     6361 tctttcatcg agaggatgac caacttcgat aagaaccttc caaacgagaa ggtgctccct
     6421 aagcactctt tgctctacga gtacttcacc gtgtacaacg agttgaccaa ggttaagtac
     6481 gtgaccgagg gaatgaggaa gcctgctttt ttgtcaggtg agcaaaagaa ggctatcgtt
     6541 gatctcttgt tcaagaccaa cagaaaggtg accgtgaagc agctcaaaga ggattacttc
     6601 aagaaaatcg agtgcttcga ttcagttgag atttctggtg ttgaggatag gttcaacgca
     6661 tctctcggaa cctaccacga tctcctcaag atcattaagg ataaggattt cttggataac
     6721 gaggaaaacg aggatatctt ggaggatatc gttcttaccc tcaccctctt tgaagataga
     6781 gagatgattg aagaaaggct caagacctac gctcatctct tcgatgataa ggtgatgaag
     6841 cagttgaaga gaagaagata cactggttgg ggaaggctct caagaaagct cattaacgga
     6901 atcagggata agcagtctgg aaagacaatc cttgatttcc tcaagtctga tggattcgct
     6961 aacagaaact tcatgcagct catccacgat gattctctca cctttaaaga ggatatccag
     7021 aaggctcagg tttcaggaca gggtgatagt ctccatgagc atatcgctaa cctcgctgga
     7081 tctcctgcaa tcaagaaggg aatcctccag actgtgaagg ttgtggatga gttggtgaag
     7141 gtgatgggaa ggcataagcc tgagaacatc gtgatcgaaa tggctagaga gaaccagacc
     7201 actcagaagg gacagaagaa ctctagggaa aggatgaaga ggatcgagga aggtatcaaa
     7261 gagcttggat ctcagatcct caaagagcac cctgttgaga acactcagct ccagaatgag
     7321 aagctctacc tctactacct ccagaacgga agggatatgt atgtggatca agagttggat
     7381 atcaacaggc tctctgatta cgatgttgat catatcgtgc cacagtcatt cttgaaggat
     7441 gattctatcg ataacaaggt gctcaccagg tctgataaga acaggggtaa gagtgataac
     7501 gtgccaagtg aagaggttgt gaagaaaatg aagaactatt ggaggcagct cctcaacgct
     7561 aagctcatca ctcagagaaa gttcgataac ttgactaagg ctgagagggg aggactctct
     7621 gaattggata aggcaggatt catcaagagg cagcttgtgg aaaccaggca gatcactaag
     7681 cacgttgcac agatcctcga ttctaggatg aacaccaagt acgatgagaa cgataagttg
     7741 atcagggaag tgaaggttat caccctcaag tcaaagctcg tgtctgattt cagaaaggat
     7801 ttccaattct acaaggtgag ggaaatcaac aactaccacc acgctcacga tgcttacctt
     7861 aacgctgttg ttggaaccgc tctcatcaag aagtatccta agctcgagtc agagttcgtg
     7921 tacggtgatt acaaggtgta cgatgtgagg aagatgatcg ctaagtctga gcaagagatc
     7981 ggaaaggcta ccgctaagta tttcttctac tctaacatca tgaatttctt caagaccgag
     8041 attaccctcg ctaacggtga gatcagaaag aggccactca tcgagacaaa cggtgaaaca
     8101 ggtgagatcg tgtgggataa gggaagggat ttcgctaccg ttagaaaggt gctctctatg
     8161 ccacaggtga acatcgttaa gaaaaccgag gtgcagaccg gtggattctc taaagagtct
     8221 atcctcccta agaggaactc tgataagctc attgctagga agaaggattg ggaccctaag
     8281 aaatacggtg gtttcgattc tcctaccgtg gcttactctg ttctcgttgt ggctaaggtt
     8341 gagaagggaa agagtaagaa gctcaagtct gttaaggaac ttctcggaat cactatcatg
     8401 gaaaggtcat ctttcgagaa gaacccaatc gatttcctcg aggctaaggg atacaaagag
     8461 gttaagaagg atctcatcat caagctccca aagtactcac tcttcgaact cgagaacggt
     8521 agaaagagga tgctcgcttc tgctggtgag cttcaaaagg gaaacgagct tgctctccca
     8581 tctaagtacg ttaactttct ttacctcgct tctcactacg agaagttgaa gggatctcca
     8641 gaagataacg agcagaagca acttttcgtt gagcagcaca agcactactt ggatgagatc
     8701 atcgagcaga tctctgagtt ctctaaaagg gtgatcctcg ctgatgcaaa cctcgataag
     8761 gtgttgtctg cttacaacaa gcacagagat aagcctatca gggaacaggc agagaacatc
     8821 atccatctct tcacccttac caacctcggt gctcctgctg ctttcaagta cttcgataca
     8881 accatcgata ggaagagata cacctctacc aaagaagtgc tcgatgctac cctcatccat
     8941 cagtctatca ctggactcta cgagactagg atcgatctct cacagctcgg tggtgattca
     9001 agggctgatc ctaagaagaa gaggaaggtt tgactcgaga tatgaagatg aagatgaaat
     9061 atttggtgtg tcaaataaaa agcttgtgtg cttaagtttg tgtttttttc ttggcttgtt
     9121 gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatcaca ttataatgaa
     9181 taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct gcagcatata
     9241 actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg agctccggtg
     9301 acggacggcg cgctggcaga catactgtcc cacaaatgaa gatggaatct gtaaaagaaa
     9361 acgcgtgaaa taatgcgtct gacaaaggtt aggtcggctg cctttaatca ataccaaagt
     9421 ggtccctacc acgatggaaa aactgtgcag tcggtttggc tttttctgac gaacaaataa
     9481 gattcgtggc cgacaggtgg gggtccacca tgtgaaggca tcttcagact ccaataatgg
     9541 agcaatgacg taagggctta cgaaataagt aagggtagtt tgggaaatgt ccactcaccc
     9601 gtcagtctat aaatacttag cccctccctc attgttaagg gagcaaaatc tcagagagat
     9661 agtcctagag agagaaagag agcaagtagc ctagaagtag tcaaggcggc gaagtattca
     9721 ggcacgtggc caggaagaag aaaagccaag acgacgaaaa caggtaagag ctaagcttcc
     9781 tgcaggttca ctgccgtata ggcagtaaaa tacacaaaag catgggtttt agagctagaa
     9841 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     9901 cgttcactgc cgtataggca gtggaacgtc ccaatactca cgttttagag ctagaaatag
     9961 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcgtt
    10021 cactgccgta taggcaggat agagagattc tatcggggtt ttagagctag aaatagcaag
    10081 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcgttcact
    10141 gccgtatagg cagttgattt gaggacaaga ggggttttag agctagaaat agcaagttaa
    10201 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgcg ttcactgccg
    10261 tataggcagg gtttggactt tgcatgtggg ttttagagct agaaatagca agttaaaata
    10321 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcgttca ctgccgtata
    10381 ggcagcaaag attgctgcta cttgggtttt agagctagaa atagcaagtt aaaataaggc
    10441 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cgttcactgc cgtataggca
    10501 ggtcgatcga caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga
    10561 attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
    10621 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
    10681 actaaaatcc agatcccccg aattaccggt ggcactggcc gtcgttttac aacgtcgtga
    10741 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
    10801 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
    10861 tggcgaatgc tagagcagct tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa
    10921 actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta
    10981 gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
    11041 atgccaacca cagggttccc ctcgggatca aagtactttg atccaacccc tccgctgcta
    11101 tagtgcagtc ggcttctgac gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa
    11161 gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg
    11221 ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg ccatgaacaa
    11281 gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc aggacttgac
    11341 caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg agaagatcac
    11401 cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac gccctggcga
    11461 cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc tactggacat
    11521 tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga
    11581 caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg ccgagttcga
    11641 gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg cccgaggcgt
    11701 gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc gcgagctgat
    11761 cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac
    11821 cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca ggcggcgcgg
    11881 tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg agaatgaacg
    11941 ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt tttcattacc
    12001 gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacggc
    12061 tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg
    12121 ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt
    12181 gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa
    12241 atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca
    12301 agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt
    12361 tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac
    12421 cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc
    12481 ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga
    12541 tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca
    12601 ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag
    12661 cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg
    12721 cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc
    12781 caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc
    12841 gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa
    12901 agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag
    12961 caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca
    13021 gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat
    13081 taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa
    13141 tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca
    13201 ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct
    13261 gggttgcctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag
    13321 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga
    13381 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg
    13441 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc
    13501 cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc
    13561 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg
    13621 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca
    13681 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact
    13741 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa
    13801 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga
    13861 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt
    13921 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga
    13981 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga
    14041 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct
    14101 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct
    14161 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg
    14221 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc
    14281 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt
    14341 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca
    14401 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggcctct ttcctgtgga
    14461 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa
    14521 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa
    14581 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc
    14641 ctgtgcataa ctgtctggcc agcgcacagc ccaagagctg caaaaagcgc ctacccttcg
    14701 gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc
    14761 aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc
    14821 actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg
    14881 aaaacctctg acacatgcag ctcccggaaa cggtcacagc ttgtctgtaa gcggatgccg
    14941 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
    15001 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
    15061 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
    15121 ataccgcatc aggccctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
    15181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
    15241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
    15301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
    15361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
    15421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
    15481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
    15541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
    15601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
    15661 actggcagca gccactggta acaggattag cagagcgagg tat
//
