LOCUS       sequence-322116-       10784 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenGate (MoClo) Level 1 Position 2 OsUbiP: Cas9: NosT cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10784)
  AUTHORS   Smedley MA, Hayta S, Clarke M, Harwood WA
  TITLE     CRISPR-Cas9 Based Genome Editing in Wheat.
  JOURNAL   Curr Protoc. 2021 Mar;1(3):e65. doi: 10.1002/cpz1.65.
  PUBMED    33687760
REFERENCE   2  (bases 1 to 10784)
  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: "Curr 
            Protoc."; date: "2021-03"; volume: "1(3):e65. doi"; pages: " 
            10.1002/cpz1.65"
COMMENT     Sequence Label: Level 1 P2 OsUbiP:Cas9:NosT
FEATURES             Location/Qualifiers
     source          1..10784
                     /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             6378..10481
                     /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             6378..10481
                     /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             6378..10481
                     /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             10494..10514
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      10530..10782
                     /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 ttggaggatg
     4381 tgaagaacag gtaaatcacg cagaagaacc catctctgat agcagctatc gattagaaca
     4441 acgaatccat attgggtccg tgggaaatac ttactgcaca ggaagggggg cgatctgacg
     4501 aggccccgcc accggcctcg acccgaggcc gaggccgacg aagcgccggc gagtacggcg
     4561 ccgcggcggc ctctgcccgt gccctctgcg cgtgggaggg agaggccgcg gtggtggggg
     4621 cgcgcgcgcg cgcgcgcgca gctggtgcgg cggcgcgggg gtcagccgcc gagccggcgg
     4681 cgacggagga gcagggcggc gtggacgcga acttccgatc ggttggtcag agtgcgcgag
     4741 ttgggcttag ccaattaggt gtcaacaatc tattgggccg taaaattcat gggccctggt
     4801 ttgtctaggc ccaatatccc gttcatttca gcccacaaat atttccccag aggattatta
     4861 aggcccacac gcagcttata gcagatcaag tacgatgttt cctgatcgtt ggatcggaaa
     4921 cgtacggtct tgatcaggca tgccgacttc gtcaaagaga ggcggcatga cctgacgcgg
     4981 agttggttcc gggcaccgtc tggatggtcg taccgggacc ggacacgtgt cgcgcctcca
     5041 actacatgga cacgtgtggt gctgccattg ggccgtacgc gtggcggtga ccgcaccgga
     5101 tgctgcctcg caccgccttg cccacgcttt atatagagag gttttctctc cattaatcgc
     5161 atagcgagtc gaatcgaccg aaggggaggg ggagcgaagc tttgcgttct ctaatcgcct
     5221 cgtcaaggta actaatcaat cacctcgtcc taatcctcga atctctcgtg gtgcccgtct
     5281 aatctcgcga ttttgatgct cgtggtggaa agcgtaggag gatcccgtgc gagttagtct
     5341 caatctctca gggtttcgtg cgattttagg gtgatccacc tcttaatcga gttacggttt
     5401 cgtgcgattt tagggtaatc ctcttaatct ctcattgatt tagggtttcg tgagaatcga
     5461 ggtagggatc tgtgttattt atatcgatct aatagatgga ttggttttga gattgttctg
     5521 tcagatgggg attgtttcga tatattaccc taatgatgtg tcagatgggg attgtttcga
     5581 tatattaccc taatgatgtg tcagatgggg attgtttcga tatattaccc taatgatgga
     5641 taataagagt agttcacagt tatgttttga tcctgccaca tagtttgagt tttgtgatca
     5701 gatttagttt tacttatttg tgcttagttc ggatgggatt gttctgatat tgttccaata
     5761 gatgaatagc tcgttaggtt aaaatcttta ggttgagtta ggcgacacat agtttatttc
     5821 ctctggattt ggattggaat tgtgttctta gtttttttcc cctggatttg gattggaatt
     5881 gtgtggagct gggttagaga attacatctg tatcgtgtac acctacttga actgtagagc
     5941 ttgggttcta aggtcaattt aatctgtatt gtatctggct ctttgcctag ttgaactgta
     6001 gtgctgatgt tgtactgtgt ttttttaccc gttttatttg ctttactcgt gcaaatcaaa
     6061 tctgtcagat gctagaacta ggtggcttta ttctgtgttc ttacatagat ctgttgtcct
     6121 gtagttactt atgtcagttt tgttattatc tgaagatatt tttggttgtt gcttgttgat
     6181 gtggtgtgag ctgtgagcag cgctcttatg attaatgatg ctgtccaatt gtagtgtagt
     6241 atgatgtgat tgatatgttc atctattttg agctgacagt accgatatcg taggatctgg
     6301 tgccaactta ttctccagct gctttttttt acctatgtta attccaatcc tttcttgcct
     6361 cttccagatc cagataaatg gacaagaagt actccattgg gctcgatatc ggcacaaaca
     6421 gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc
     6481 tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc ctgttcgact
     6541 ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga tatacccgca
     6601 gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct aaggtggatg
     6661 actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa aagcacgagc
     6721 gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag tacccaacca
     6781 tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg cggttgatct
     6841 atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag ggggacctga
     6901 acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact tacaatcagc
     6961 ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc ctgagcgcta
     7021 ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga
     7081 acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac tttaaatcta
     7141 acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac gatgatgatc
     7201 tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg gcggcaaaga
     7261 acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag atcaccaaag
     7321 ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac ttgactttgc
     7381 tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc ttcgatcagt
     7441 ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa ttttacaaat
     7501 ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta aagcttaaca
     7561 gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc caccagattc
     7621 acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc tttttgaaag
     7681 ataacaggga aaagattgag aaaatcctca catttcggat accctactat gtaggccccc
     7741 tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag actatcactc
     7801 cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc atcgaaagga
     7861 tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt
     7921 acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca gaagggatga
     7981 gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc ctcttcaaga
     8041 cgaaccggaa agttaccgtg aaacagctca aagaagatta tttcaaaaag attgaatgtt
     8101 tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc
     8161 acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag aacgaggaca
     8221 ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg attgaagaac
     8281 gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc
     8341 gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga gacaagcaga
     8401 gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg aacttcatgc
     8461 agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca caagtttctg
     8521 gccaggggga cagtctccac gagcacatcg ctaatcttgc aggtagccca gctatcaaaa
     8581 agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata
     8641 agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag aagggacaga
     8701 agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg gggtcccaaa
     8761 tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc tacctgtact
     8821 acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat cggctctccg
     8881 actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct attgataata
     8941 aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc tcagaagaag
     9001 ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg atcacacaac
     9061 ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg gataaagccg
     9121 gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg gcccaaattc
     9181 tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga gaggtgaaag
     9241 ttattactct gaagtctaag ctggtttcag atttcagaaa ggactttcag ttttataagg
     9301 tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca gtggtaggca
     9361 ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga gactataaag
     9421 tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag gccaccgcta
     9481 agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca ctggccaatg
     9541 gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa atcgtgtggg
     9601 acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag gtgaacatcg
     9661 ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc ccgaaaagga
     9721 acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac ggcggattcg
     9781 attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa gggaagtcta
     9841 aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga tcaagcttcg
     9901 aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa aaagacctca
     9961 tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa cgaatgctcg
    10021 ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa tacgttaatt
    10081 tcttgtatct ggccagccac tatgaaaagc tcaaaggatc tcccgaagat aatgagcaga
    10141 agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag caaataagcg
    10201 aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt tctgcttaca
    10261 ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac ttgtttactc
    10321 tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata gacagaaagc
    10381 ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca attacggggc
    10441 tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct gaccccaaga
    10501 agaagaggaa ggtgtgagct tgtcaagcag atcgttcaaa catttggcaa taaagtttct
    10561 taagattgaa tcctgttgcc ggtcttgcga tgattatcat ataatttctg ttgaattacg
    10621 ttaagcatgt aataattaac atgtaatgca tgacgttatt tatgagatgg gtttttatga
    10681 ttagagtccc gcaattatac atttaatacg cgatagaaaa caaaatatag cgcgcaaact
    10741 aggataaatt atcgcgcgcg gtgtcatcta tgttactaga tcga
//
