LOCUS       sequence-119203-       10772 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 Golden Gate Cassette: Cas9 expression cassette for 
            monocotyledonous plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10772)
  AUTHORS   Lawrenson T, Shorinola O, Stacey N, Li C, Ostergaard L, Patron N, 
            Uauy C, Harwood W
  TITLE     Induction of targeted, heritable mutations in barley and Brassica 
            oleracea using RNA-guided Cas9 nuclease.
  JOURNAL   Genome Biol. 2015 Nov 30;16(1):258. doi: 10.1186/s13059-015-0826-7.
  PUBMED    26616834
REFERENCE   2  (bases 1 to 10772)
  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: "Genome 
            Biol."; date: "2015-11-30"; volume: "16(1):258. doi"; pages: " 
            10.1186/s13059-015-0826-7"
COMMENT     Sequence Label: pICSL11056
FEATURES             Location/Qualifiers
     source          1..10772
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    46..70
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(226..1374)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(1710..2298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1790..1809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2469..3260,3261..3329))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3092..3111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3330..3434)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3521..4152
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4244..4268
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6366..10469
                     /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             6366..10469
                     /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             6366..10469
                     /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             10482..10502
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      10518..10770
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 cgctactatt gtcttctgca cgaagtggtt taaactatca gtgtttgaca ggatatattg
       61 gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg
      121 aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt ccccagatca
      181 ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc gtttgcaatg
      241 caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca actcttcgca
      301 ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc cgcacatgag
      361 gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat
      421 ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg tgtagtggtc
      481 gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg acgttttctt
      541 gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt gcccaacgcg
      601 gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct cggccttcgt
      661 gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga acgtgaaggt
      721 gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc acaccagttc
      781 gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg
      841 gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag
      901 ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc
      961 gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg
     1021 cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt
     1081 catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct cctgttcaag
     1141 acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag ccagttgcac
     1201 gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg actggaaggt
     1261 ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa
     1321 ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat tcattcaccc
     1381 tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc
     1441 gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg tcccgtagac
     1501 cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga ataccagcga
     1561 ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact tgatgcggaa
     1621 gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg atctgccagg
     1681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2581 tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca
     2641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3361 tatcaaggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     3421 ataggggttc cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg ccgttcgcgg
     3481 ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg gagggttcga
     3541 gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta
     3601 aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta gcggtggccg
     3661 aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag cccctcaaat
     3721 gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc
     3781 cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact tatccccagg
     3841 cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag
     3901 gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt caacgccgcg
     3961 ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt gagggccaag
     4021 ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg cttcgacggc
     4081 gtttctggcg cgtttgcagg gccatagacg gccgccagcc cagcggcgag ggcaaccagc
     4141 ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg gctgcctgta tcgagtggtg
     4201 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
     4261 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
     4321 ggggtggatg cagtgggccc cactctgtga agacaagcaa gaattcaagc ttggaggtgc
     4381 agcgtgaccc ggtcgtgccc ctctctagag ataatgagca ttgcatgtct aagttataaa
     4441 aaattaccac atattttttt tgtcacactt gtttgaagtg cagtttatct atctttatac
     4501 atatatttaa actttactct acgaataata taatctatag tactacaata atatcagtgt
     4561 tttagagaat catataaatg aacagttaga catggtctaa aggacaattg agtattttga
     4621 caacaggact ctacagtttt atctttttag tgtgcatgtg ttctcctttt tttttgcaaa
     4681 tagcttcacc tatataatac ttcatccatt ttattagtac atccatttag ggtttagggt
     4741 taatggtttt tatagactaa tttttttagt acatctattt tattctattt tagcctctaa
     4801 attaagaaaa ctaaaactct attttagttt ttttatttaa taatttagat ataaaataga
     4861 ataaaataaa gtgactaaaa attaaacaaa taccctttaa gaaattaaaa aaactaagga
     4921 aacatttttc ttgtttcgag tagataatgc cagcctgtta aacgccgtcg acgagtctaa
     4981 cggacaccaa ccagcgaacc agcagcgtcg cgtcgggcca agcgaagcag acggcacggc
     5041 atctctgtcg ctgcctctgg acccctctcg agagttccgc tccaccgttg gacttgctcc
     5101 gctgtcggca tccagaaatt gcgtggcgga gcggcagacg tgagccggca cggcaggcgg
     5161 cctcctcctc ctctcacggc acggcagcta cgggggattc ctttcccacc gctccttcgc
     5221 tttcccttcc tcgcccgccg taataaatag acaccccctc cacaccctct ttccccaacc
     5281 tcgtgttgtt cggagcgcac acacacacaa ccagatctcc cccaaatcca cccgtcggca
     5341 cctccgcttc aaggtacgcc gctcgtcctc cccccccccc cctctctacc ttctctagat
     5401 cggcgttccg gtccatggtt agggcccggt agttctactt ctgttcatgt ttgtgttaga
     5461 tccgtgtttg tgttagatcc gtgctgctag cgttcgtaca cggatgcgac ctgtacgtca
     5521 gacacgttct gattgctaac ttgccagtgt ttctctttgg ggaatcctgg gatggctcta
     5581 gccgttccgc agacgggatc gatttcatga ttttttttgt ttcgttgcat agggtttggt
     5641 ttgccctttt cctttatttc aatatatgcc gtgcacttgt ttgtcgggtc atcttttcat
     5701 gctttttttt gtcttggttg tgatgatgtg gtctggttgg gcggtcgttc tagatcggag
     5761 tagaattctg tttcaaacta cctggtggat ttattaattt tggatctgta tgtgtgtgcc
     5821 atacatattc atagttacga attgaagatg atggatggaa atatcgatct aggataggta
     5881 tacatgttga tgcgggtttt actgatgcat atacagagat gctttttgtt cgcttggttg
     5941 tgatgatgtg gtgtggttgg gcggtcgttc attcgttcta gatcggagta gaatactgtt
     6001 tcaaactacc tggtgtattt attaattttg gaactgtatg tgtgtgtcat acatcttcat
     6061 agttacgagt ttaagatgga tggaaatatc gatctaggat aggtatacat gttgatgtgg
     6121 gttttactga tgcatataca tgatggcata tgcagcatct attcatatgc tctaaccttg
     6181 agtacctatc tattataata aacaagtatg ttttataatt attttgatct tgatatactt
     6241 ggatgatggc atatgcagca gctatatgtg gattttttta gccctgcctt catacgctat
     6301 ttatttgctt ggtactgttt cttttgtcga tgctcaccct gttgtttggt gttacttctg
     6361 caggaatgga caagaagtac tccattgggc tcgatatcgg cacaaacagc gtcggctggg
     6421 ccgtcattac ggacgagtac aaggtgccga gcaaaaaatt caaagttctg ggcaataccg
     6481 atcgccacag cataaagaag aacctcattg gcgccctcct gttcgactcc ggggagacgg
     6541 ccgaagccac gcggctcaaa agaacagcac ggcgcagata tacccgcaga aagaatcgga
     6601 tctgctacct gcaggagatc tttagtaatg agatggctaa ggtggatgac tctttcttcc
     6661 ataggctgga ggagtccttt ttggtggagg aggataaaaa gcacgagcgc cacccaatct
     6721 ttggcaatat cgtggacgag gtggcgtacc atgaaaagta cccaaccata tatcatctga
     6781 ggaagaagct tgtagacagt actgataagg ctgacttgcg gttgatctat ctcgcgctgg
     6841 cgcatatgat caaatttcgg ggacacttcc tcatcgaggg ggacctgaac ccagacaaca
     6901 gcgatgtcga caaactcttt atccaactgg ttcagactta caatcagctt ttcgaagaga
     6961 acccgatcaa cgcatccgga gttgacgcca aagcaatcct gagcgctagg ctgtccaaat
     7021 cccggcggct cgaaaacctc atcgcacagc tccctgggga gaagaagaac ggcctgtttg
     7081 gtaatcttat cgccctgtca ctcgggctga cccccaactt taaatctaac ttcgacctgg
     7141 ccgaagatgc caagcttcaa ctgagcaaag acacctacga tgatgatctc gacaatctgc
     7201 tggcccagat cggcgaccag tacgcagacc tttttttggc ggcaaagaac ctgtcagacg
     7261 ccattctgct gagtgatatt ctgcgagtga acacggagat caccaaagct ccgctgagcg
     7321 ctagtatgat caagcgctat gatgagcacc accaagactt gactttgctg aaggcccttg
     7381 tcagacagca actgcctgag aagtacaagg aaattttctt cgatcagtct aaaaatggct
     7441 acgccggata cattgacggc ggagcaagcc aggaggaatt ttacaaattt attaagccca
     7501 tcttggaaaa aatggacggc accgaggagc tgctggtaaa gcttaacaga gaagatctgt
     7561 tgcgcaaaca gcgcactttc gacaatggaa gcatccccca ccagattcac ctgggcgaac
     7621 tgcacgctat cctcaggcgg caagaggatt tctacccctt tttgaaagat aacagggaaa
     7681 agattgagaa aatcctcaca tttcggatac cctactatgt aggccccctc gcccggggaa
     7741 attccagatt cgcgtggatg actcgcaaat cagaagagac tatcactccc tggaacttcg
     7801 aggaagtcgt ggataagggg gcctctgccc agtccttcat cgaaaggatg actaactttg
     7861 ataaaaatct gcctaacgaa aaggtgcttc ctaaacactc tctgctgtac gagtacttca
     7921 cagtttataa cgagctcacc aaggtcaaat acgtcacaga agggatgaga aagccagcat
     7981 tcctgtctgg agagcagaag aaagctatcg tggacctcct cttcaagacg aaccggaaag
     8041 ttaccgtgaa acagctcaaa gaagattatt tcaaaaagat tgaatgtttc gactctgttg
     8101 aaatcagcgg agtggaggat cgcttcaacg catccctggg aacgtatcac gatctcctga
     8161 aaatcattaa agacaaggac ttcctggaca atgaggagaa cgaggacatt cttgaggaca
     8221 ttgtcctcac ccttacgttg tttgaagata gggagatgat tgaagaacgc ttgaaaactt
     8281 acgctcatct cttcgacgac aaagtcatga aacagctcaa gaggcgccga tatacaggat
     8341 gggggcggct gtcaagaaaa ctgatcaatg ggatccgaga caagcagagt ggaaagacaa
     8401 tcctggattt tcttaagtcc gatggatttg ccaaccggaa cttcatgcag ttgatccatg
     8461 atgactctct cacctttaag gaggacatcc agaaagcaca agtttctggc cagggggaca
     8521 gtctccacga gcacatcgct aatcttgcag gtagcccagc tatcaaaaag ggaatactgc
     8581 agaccgttaa ggtcgtggat gaactcgtca aagtaatggg aaggcataag cccgagaata
     8641 tcgttatcga gatggcccga gagaaccaaa ctacccagaa gggacagaag aacagtaggg
     8701 aaaggatgaa gaggattgaa gagggtataa aagaactggg gtcccaaatc cttaaggaac
     8761 acccagttga aaacacccag cttcagaatg agaagctcta cctgtactac ctgcagaacg
     8821 gcagggacat gtacgtggat caggaactgg acatcaatcg gctctccgac tacgacgtgg
     8881 atcatatcgt gccccagtct tttctcaaag atgattctat tgataataaa gtgttgacaa
     8941 gatccgataa aaatagaggg aagagtgata acgtcccctc agaagaagtt gtcaagaaaa
     9001 tgaaaaatta ttggcggcag ctgctgaacg ccaaactgat cacacaacgg aagttcgata
     9061 atctgactaa ggctgaacga ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa
     9121 ggcagcttgt tgagacacgc cagatcacca agcacgtggc ccaaattctc gattcacgca
     9181 tgaacaccaa gtacgatgaa aatgacaaac tgattcgaga ggtgaaagtt attactctga
     9241 agtctaagct ggtttcagat ttcagaaagg actttcagtt ttataaggtg agagagatca
     9301 acaattacca ccatgcgcat gatgcctacc tgaatgcagt ggtaggcact gcacttatca
     9361 aaaaatatcc caagcttgaa tctgaatttg tttacggaga ctataaagtg tacgatgtta
     9421 ggaaaatgat cgcaaagtct gagcaggaaa taggcaaggc caccgctaag tacttctttt
     9481 acagcaatat tatgaatttt ttcaagaccg agattacact ggccaatgga gagattcgga
     9541 agcgaccact tatcgaaaca aacggagaaa caggagaaat cgtgtgggac aagggtaggg
     9601 atttcgcgac agtccggaag gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg
     9661 aagtacagac cggaggcttc tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc
     9721 tgatcgcacg caaaaaagat tgggacccca agaaatacgg cggattcgat tctcctacag
     9781 tcgcttacag tgtactggtt gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa
     9841 gcgtcaagga actgctgggc atcacaatca tggagcgatc aagcttcgaa aaaaacccca
     9901 tcgactttct cgaggcgaaa ggatataaag aggtcaaaaa agacctcatc attaagcttc
     9961 ccaagtactc tctctttgag cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg
    10021 agctgcagaa aggtaacgag ctggcactgc cctctaaata cgttaatttc ttgtatctgg
    10081 ccagccacta tgaaaagctc aaaggatctc ccgaagataa tgagcagaag cagctgttcg
    10141 tggaacaaca caaacactac cttgatgaga tcatcgagca aataagcgaa ttctccaaaa
    10201 gagtgatcct cgccgacgct aacctcgata aggtgctttc tgcttacaat aagcacaggg
    10261 ataagcccat cagggagcag gcagaaaaca ttatccactt gtttactctg accaacttgg
    10321 gcgcgcctgc agccttcaag tacttcgaca ccaccataga cagaaagcgg tacacctcta
    10381 caaaggaggt cctggacgcc acactgattc atcagtcaat tacggggctc tatgaaacaa
    10441 gaatcgacct ctctcagctc ggtggagaca gcagggctga ccccaagaag aagaggaagg
    10501 tgtgagcttg tcaagcagat cgttcaaaca tttggcaata aagtttctta agattgaatc
    10561 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
    10621 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
    10681 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
    10741 cgcgcgcggt gtcatctatg ttactagatc ga
//
