LOCUS       sequence-182722-       16103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T-DNA vector for targeted deletion of 58kb region in Medicago 
            truncatula (tRNA 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 16103)
  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 16103)
  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: pSC4
FEATURES             Location/Qualifiers
     source          1..16103
                     /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)"
     misc_RNA        9884..9959
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        10057..10132
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        10230..10305
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        10403..10478
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        10576..10651
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        10749..10824
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     polyA_signal    10921..11095
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(11114..11131)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11114..11130)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11123..11145)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    11333..11357
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             12658..13287
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             13716..14789
                     /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      14855..15049
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(15285..15307)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    15393..15533
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     15448..15467
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(15548..15565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(15719..16103,1..204))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15799..15818)
                     /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 tgcaggaaca aagcaccagt ggtctagtgg tagaatagta ccctgccacg gtacagaccc
     9841 gggttcgatt cccggctggt gcataaaata cacaaaagca tgggttttag agctagaaat
     9901 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgca
     9961 acaaagcacc agtggtctag tggtagaata gtaccctgcc acggtacaga cccgggttcg
    10021 attcccggct ggtgcatgga acgtcccaat actcacgttt tagagctaga aatagcaagt
    10081 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcaacaaagc
    10141 accagtggtc tagtggtaga atagtaccct gccacggtac agacccgggt tcgattcccg
    10201 gctggtgcag atagagagat tctatcgggg ttttagagct agaaatagca agttaaaata
    10261 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcaacaa agcaccagtg
    10321 gtctagtggt agaatagtac cctgccacgg tacagacccg ggttcgattc ccggctggtg
    10381 cattgatttg aggacaagag gggttttaga gctagaaata gcaagttaaa ataaggctag
    10441 tccgttatca acttgaaaaa gtggcaccga gtcggtgcaa caaagcacca gtggtctagt
    10501 ggtagaatag taccctgcca cggtacagac ccgggttcga ttcccggctg gtgcaggttt
    10561 ggactttgca tgtgggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
    10621 tcaacttgaa aaagtggcac cgagtcggtg caacaaagca ccagtggtct agtggtagaa
    10681 tagtaccctg ccacggtaca gacccgggtt cgattcccgg ctggtgcaca aagattgctg
    10741 ctacttgggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
    10801 gaaaaagtgg caccgagtcg gtgcaacaaa gcaccagtgg tctagtggta gaatagtacc
    10861 ctgccacggt acagacccgg gttagattcc cggctggtgc agtcgatcga caagctcgag
    10921 tttctccata ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt
    10981 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
    11041 ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcccccg
    11101 aattaccggt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
    11161 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
    11221 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc tagagcagct
    11281 tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa actatcagtg tttgacagga
    11341 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
    11401 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
    11461 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
    11521 gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
    11581 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
    11641 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
    11701 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
    11761 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
    11821 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
    11881 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
    11941 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
    12001 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
    12061 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
    12121 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
    12181 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
    12241 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
    12301 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
    12361 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
    12421 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacggc tcaaccgtgc ggctgcatga
    12481 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
    12541 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
    12601 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
    12661 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
    12721 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
    12781 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
    12841 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
    12901 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
    12961 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
    13021 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
    13081 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
    13141 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
    13201 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
    13261 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
    13321 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
    13381 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
    13441 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
    13501 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
    13561 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
    13621 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgcctg ccggccctgc
    13681 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
    13741 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
    13801 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
    13861 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
    13921 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
    13981 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
    14041 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
    14101 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
    14161 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
    14221 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
    14281 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
    14341 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
    14401 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
    14461 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
    14521 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
    14581 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
    14641 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
    14701 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
    14761 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
    14821 tagcagggga aaaaggtcga aaaggcctct ttcctgtgga tagcacgtac attgggaacc
    14881 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
    14941 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
    15001 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
    15061 agcgcacagc ccaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
    15121 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
    15181 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
    15241 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
    15301 ctcccggaaa cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
    15361 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
    15421 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
    15481 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggccctctt
    15541 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    15601 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    15661 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    15721 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    15781 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    15841 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    15901 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    15961 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    16021 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    16081 acaggattag cagagcgagg tat
//
