LOCUS       sequence-444273-       17577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for Aspergillus CRISPR-Cas9 genetic engineering with Golden 
            Gate cloning drop-out cassette for protospacer insertion and ble 
            selectable marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17577)
  AUTHORS   Modaffari D, Finlayson A, Miao Y, Wallace EWJ, Sawin KE
  TITLE     Improved gene editing and fluorescent-protein tagging in Aspergillus
            nidulans using a Golden Gate-based CRISPR-Cas9 plasmid system.
  JOURNAL   Wellcome Open Res. 2024 Oct 17;9:602. doi: 
            10.12688/wellcomeopenres.23086.1. eCollection 2024.
  PUBMED    39640368
REFERENCE   2  (bases 1 to 17577)
  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: "Wellcome 
            Open Res."; date: "2024-10-17"; volume: "9:602. doi"; pages: " 
            10.12688/wellcomeopenres.23086.1. eCollection 2024"
COMMENT     Sequence Label: pDM030
FEATURES             Location/Qualifiers
     source          1..17577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(63..96)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(566..582)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(681..698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(852..1440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(932..951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1564..2355,2356..2424))
                     /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     2187..2206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2425..2516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2558..2577
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2583..2602
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    2809..2842
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      3155..8402
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     ncRNA           9111..9178
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     RBS             9338..9349
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             9356..10069
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      10081..10152
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      10168..10195
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        10213..10288
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             11494..15597
                     /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             11494..15597
                     /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             15601..15621
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             16443..16817
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
ORIGIN
        1 aggtaaatcc ccactaccgc attaagacct cagcgcggcc gcaaatttaa ataaaatgaa
       61 gtgaagttcc tatactttct agagaatagg aacttctata gtgagtcgaa taagggcgac
      121 acaaaattta ttctaaatgc ataataaata ctgataacat cttatagttt gtattatatt
      181 ttgtattatc gttgacatgt ataattttga tatcaaaaac tgattttccc tttattattt
      241 tcgagattta ttttcttaat tctctttaac aaactagaaa tattgtatat acaaaaaatc
      301 ataaataata gatgaatagt ttaattatag gtgttcatca atcgaaaaag caacgtatct
      361 tatttaaagt gcgttgcttt tttctcattt ataaggttaa ataattctca tatatcaagc
      421 aaagtgacag gcgcccttaa atattctgac aaatgctctt tccctaaact ccccccataa
      481 aaaaacccgc cgaagcgggt ttttacgtta tttgcggatt aacgattact cgttatcaga
      541 accgcccagg gggcccgagc ttaagactgg ccgtcgtttt acaacacaga aagagtttgt
      601 agaaacgcaa aaaggccatc cgtcaggggc cttctgctta gtttgatgcc tggcagttcc
      661 ctactctcgc cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
      721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      781 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1021 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1261 aagtggtggg ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgacgc gcgcgtaact
     1501 cacgttaagg gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg
     1561 cttttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     1621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1681 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
     1741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1861 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     2041 actcatggtt atggcagcgc tacataattc tcttactgtc atgccatccg taagatgctt
     2101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     2161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     2221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     2281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     2341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     2401 gacacggaaa tgttgaatac tcatattctt cctttttcaa tattattgaa gcatttatca
     2461 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     2521 ggtcagtgtt acaaccaatt aaccaattct gaacattatc gcgagcccat ttatacctga
     2581 atatggctca taacacccct tgtttgcctg gcggcagtag cgcggtggtc ccacctgacc
     2641 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg actccccatg
     2701 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2761 tttcgcccgg gctaattatg gggtgtcgcc cttattcgac tctatagtga agttcctatt
     2821 ctctagaaag tataggaact tctgaagtgg ggatttaaat gcggccgcgc tgagggttta
     2881 atcgacgaag cagctgacgg ccagtgccaa gcttaacgcg taccgggccc agtatatgtt
     2941 ccgcagatga ctggagctct gccatacgtg ccctctcaag caccatttgt tccatctaca
     3001 gagactagtc accaactagt ctatcaagac tcacagggta cattgctgag accaactgac
     3061 cagaggcagg gtagcggatt gacggctcca tctccttcac ttacaaggtc tattgaaagc
     3121 cctttagcat caccaagcgg agaatagatt gttaagctta ttttttgtat actgttttgt
     3181 gatagcacga agtttttcca cggtatcttg ttaaaaatat atatttgtgg cgggcttacc
     3241 tacatcaaat taataagaga ctaattataa actaaacaca caagcaagct actttagggt
     3301 aaaagtttat aaatgctttt gacgtataaa cgttgcttgt atttattatt acaattaaag
     3361 gtggatagaa aacctagaga ctagttagaa actaatctca ggtttgcgtt aaactaaatc
     3421 agagcccgag aggttaacag aacctagaag gggactagat atccgggtag ggaaacaaaa
     3481 aaaaaaacaa gacagccaca tattagggag actagttaga agctagttcc aggactagga
     3541 aaataaaaga caatgatacc acagtctagt tgacaactag atagattcta gattgaggcc
     3601 aaagtctctg agatccaggt tagttgcaac taatactagt tagtatctag tctcctataa
     3661 ctctgaagct agaataactt actactatta tcctcaccac tgttcagctg cgcaaacgga
     3721 gtgattgcaa ggtgttcaga gactagttat tgactagtca gtgactagca ataactaaca
     3781 aggtattaac ctaccatgtc tgccatcacc ctgcacttcc tcgggctcag cagccttttc
     3841 ctcctcattt tcatgctcat tttccttgtt taagactgtg actagtcaaa gactagtcca
     3901 gaaccacaaa ggagaaatgt cttaccactt tcttcattgc ttgtctcttt tgcattatcc
     3961 atgtctgcaa ctagttagag tctagttagt gactagtccg acgaggactt gcttgtctcc
     4021 ggattgttgg aggaactctc cagggcctca agatccacaa cagagccttc tagaagactg
     4081 gtcaataact agttggtctt tgtctgagtc tgacttacga ggttgcatac tcgctccctt
     4141 tgcctcgtca atcgatgaga aaaagcgcca aaactcgcaa tatggctttg aaccacacgg
     4201 tgctgagact agttagaatc tagtcccaaa ctagcttgga tagcttacct ttgccctttg
     4261 cgttgcgaca ggtcttgcag ggtatggttc ctttctcacc agctgattta gctgccttgc
     4321 taccctcacg gcggatctgc ataaagagtg gctagaggtt ataaattagc actgatccta
     4381 ggtacggggc tgaatgtaac ttgcctttcc tttctcatcg cgcggcaaga caggcttgct
     4441 caaattccta ccagtcacag gggtatgcac ggcgtacgga ccacttgaac tagtcacaga
     4501 ttagttagca actagtctgc attgaatggc tgtacttacg ggccctcgcc attgtcctga
     4561 tcatttccag cttcaccctc gttgctgcaa agtagttagt gactagtcaa ggactagttg
     4621 aaatgggaga agaaactcac gaattctcga cacccttagt attgtggtcc ttggacttgg
     4681 tgctgctata tattagctaa tacactagtt agactcacag aaacttacgc agctcgcttg
     4741 cgcttcttgg taggagtcgg ggttgggaga acagtgcctt caaacaagcc ttcataccat
     4801 gctacttgac tagtcaggga ctagtcacca agtaatctag ataggacttg cctttggcct
     4861 ccatcagttc cttcatagtg ggaggtccat tgtgcaatgt aaactccatg ccgtgggagt
     4921 tcttgtcctt caagtgcttg accaatatgt ttctgttggc agagggaacc tgtcaactag
     4981 ttaataacta gtcagaaact agtatagcag tagactcact gtacgcttga ggcatccctt
     5041 cactcggcag tagacttcat atggatggat atcaggcacg ccattgtcgt cctgtggact
     5101 agtcagtaac taggcttaaa gctagtcggg tcggcttact atcttgaaat ccggcagcgt
     5161 aagctccccg tccttaactg cctcgagata gtgacagtac tctggggact ttcggagatc
     5221 gttatcgcga atgctcggca tactaatcgt tgactagtct tggactagtc ccgagcaaaa
     5281 aggattggag gaggaggagg aaggtgagag tgagacaaag agcgaaataa gagcttcaaa
     5341 ggctatctct aagcagtatg aaggttaagt atctagttct tgactagatt taaaagagat
     5401 ttcgactagt tatgtacctg gagtttggat ataggaatgt gttgtggtaa cgaaatgtaa
     5461 gggggaggaa agaaaaagtc ggtcaagagg taactctaag tcggccattc ctttttggga
     5521 ggcgctaacc ataaacggca tggtcgactt agagttagct cagggaattt agggagttat
     5581 ctgcgaccac cgaggaacgg cggaatgcca aagaatcccg atggagctct agctggcggt
     5641 tgacaacccc accttttggc gtttctgcgg cgttgcaggc gggactggat acttcgtaga
     5701 accagaaagg caaggcagaa cgcgctcagc aagagtgttg gaagtgatag catgatgtgc
     5761 cttgttaact aggtcaaaat ctgcagtatg cttgatgtta tccaaagtgt gagagaggaa
     5821 ggtccaaaca tacacgattg ggagagggcc taggtataag agtttttgag tagaacgcat
     5881 gtgagcccag ccatctcgag gagattaaac acgggccggc atttgatggc tatgttagta
     5941 ccccaatgga aacggtgaga gtccagtggt cgcagataac tccctaaatt ccctgagcta
     6001 actctaagtc gaccatgccg tttatggtta gcgcctccca aaaaggaatg gccgacttag
     6061 agttacctct tgaccgactt tttctttcct cccccttaca tttcgttacc acaacacatt
     6121 cctatatcca aactccaggt acataactag tcgaaatctc ttttaaatct agtcaagaac
     6181 tagatactta accttcatac tgcttagaga tagcctttga agctcttatt tcgctctttg
     6241 tctcactctc accttcctcc tcctcctcca atcctttttg ctcgggacta gtccaagact
     6301 agtcaacgat tagtatgccg agcattcgcg ataacgatct ccgaaagtcc ccagagtact
     6361 gtcactatct cgaggcagtt aaggacgggg agcttacgct gccggatttc aagatagtaa
     6421 gccgacccga ctagctttaa gcctagttac tgactagtcc acaggacgac aatggcgtgc
     6481 ctgatatcca tccatatgaa gtctactgcc gagtgaaggg atgcctcaag cgtacagtga
     6541 gtctactgct atactagttt ctgactagtt attaactagt tgacaggttc cctctgccaa
     6601 cagaaacata ttggtcaagc acttgaagga caagaactcc cacggcatgg agtttacatt
     6661 gcacaatgga cctcccacta tgaaggaact gatggaggcc aaaggcaagt cctatctaga
     6721 ttacttggtg actagtccct gactagtcaa gtagcatggt atgaaggctt gtttgaaggc
     6781 actgttctcc caaccccgac tcctaccaag aagcgcaagc gagctgcgta agtttccgtg
     6841 agtctaacta gtgtattagc taatatatag cagcaccaag tccaaggacc acaatactaa
     6901 gggtgtcgag aattcgtgag tttcttctcc catttcaact agtccttgac tagtcactaa
     6961 ctactttgca gcaacgaggg tgaagctgga aatgatcagg acaatggcga gggcccgtaa
     7021 gtacagccat tcaatgcaga ctagttgcta actaatctgt gactagttca agtggtccgt
     7081 acgccgtgca tacccctgtg actggtagga atttgagcaa gcctgtcttg ccgcgcgatg
     7141 agaaaggaag ggcaagttac attcagcccc gtacctagga tcagtgctaa tttataacct
     7201 ctagccactc tttatgcaga tccgccgtga gggtagcaag gcagctaaat cagctggtga
     7261 gaaaggaacc ataccctgca agacctgtcg caacgcaaag ggcaaaggta agctatccaa
     7321 gctagtttgg gactagattc taactagtct cagcaccgtg tggttcaaag ccatattgcg
     7381 agttttggcg ctttttctca tcgattgacg aggcaaaggg agcgagtatg caacctcgta
     7441 agtcagactc agacaaagac caactagtta ttgaccagtc ttctagaagg ctctgttgtg
     7501 gatcttgagg ccctggagag ttcctccaac aatccggaga caagcaagtc ctcgtcggac
     7561 tagtcactaa ctagactcta actagttgca gacatggata atgcaaaaga gacaagcaat
     7621 gaagaaagtg gtaagacatt tctcctttgt ggttctggac tagtcttcga ctagtcacag
     7681 tcttaaacaa ggaaaatgag catgaaaatg aggaggaaaa ggctgctgag cccgaggaag
     7741 tgcagggtga tggcagacat ggtaggttaa taccttgtta gttattgcta gtcactgact
     7801 agtcaataac tagtctctga acaccttgca atcactccgt ttgcgcagct gaacagtggt
     7861 gaggataata gtagtaagtt attctagctt cagagttata ggagactaga tactaactag
     7921 tattagttgc aactaacctg gatctcagag actttggcct caatctagaa tctatctagt
     7981 tgtcaactag actgtggtat cattgtcttt tattttccta gtcctggaac tagcttctaa
     8041 ctagtctccc taatatgtgg ctgtcttgtt tttttttttt gtttccctac ccggatatct
     8101 agtccccttc taggttctgt taacctctcg ggctctgatt tagtttaacg caaacctgag
     8161 attagtttct aactagtctc taggttttct atccaccttt aattgtaata ataaatacaa
     8221 gcaacgttta tacgtcaaaa gcatttataa acttttaccc taaagtagct tgcttgtgtg
     8281 tttagtttat aattagtctc ttattaattt gatgtaggta agcccgccac aaatatatat
     8341 ttttaacaag ataccgtgga aaaacttcgt gctatcacaa aacagtatac aaaaaataag
     8401 cttaacaatc tattctccgc ttggtgatgc taaagggctt tcaatagacc ttgtaagtga
     8461 aggagatgga gccgtcaatc cgctaccctg cctctggtca gttggtctca gcaatgtacc
     8521 ctgtgagtct tgatagacta gttggtgact agtctctgta gatggaacaa atggtgcttg
     8581 agagggcacg tatggcagag ctccagtcat ctgcggaaca tatactgggc ccgggaagat
     8641 ctgatcacat agatgctcgg ttgacaggac cacctatagc aagtactttg taggaggttc
     8701 tcctaacgct tggcttcaaa tccaatcagg atatcagtga gttactgttt gtcatcgcat
     8761 cctcatgaag tcgctcaaca gccactgaga agcaaatatt tgggagacca tccctgatgt
     8821 tgaaatttta cgctggagcc cattcaccgg tgagctcggg gacagtctgg ccagtggagc
     8881 ggaaaatctt caactaactc tgattggtac cacaggtagc cccacagaac aaaccgagca
     8941 aacatgaaaa ttttcgcttg aggttagcgc actcgctagc gcctgcccgc aaatataaaa
     9001 ggccccgaaa tcccatgcat tttggaaatc aagcgactct acgtatgtca tccccctccc
     9061 tacctttggg aaaacctctc aactgcaagt cagaacattt tgctaacagc ggccggcatg
     9121 gtcccagcct cctcgctggc gccggctggg caacatgctt cggcatggcg aatgggaccg
     9181 atgcaggtgg aaagtgaaac gtgatttcat gcgtcatttt gaacattttg taaatcttat
     9241 ttaataatgt gtgcggcaat tcacatttaa tttatgaatg ttttcttaac atcgcggcaa
     9301 ctcaagaaac ggcaggttcg gatcttagct actagagaaa gaggagaaat actagatgcg
     9361 taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg atggtgatgt
     9421 caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta atggtaaact
     9481 gacgctgaag ttcatctgta ctactggtaa actgccggtt ccttggccga ctctggtaac
     9541 gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga agcagcatga
     9601 cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt cctttaagga
     9661 tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc tggtaaaccg
     9721 cattgagctg aaaggcattg actttaaaga ggacggcaat atcctgggcc ataagctgga
     9781 atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa atggcattaa
     9841 agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg ctgatcacta
     9901 ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc actatctgag
     9961 cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg ttctgctgga
    10021 gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat gaccaggcat
    10081 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
    10141 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
    10201 acacctgcag tcgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
    10261 acttgaaaaa gtggcaccga gtcggtgctt ttgcatcatt ggtctagtgg tagaattcat
    10321 cgttgccatc gatgaggccc gtgttcgatt cacggatgat gcatgtcaat ttttacactt
    10381 gatgttggtg taatcaggcg atcagagttt gctcaaattc tctcttgttt tagattctga
    10441 agaatataat ttttacatga ctagttcatt ttacttccaa gtagactatt tgtagattga
    10501 gattgccgtc aactttattc atagaatcac attaaattgc tcattgacct gcatgtctat
    10561 gcacacattc acatctatct tctcagggta ttaattaaga cctcagccga gacagcagaa
    10621 tcaccgccca agttaagcct ttgtgctgat catgctctcg aacgggccaa gttcgggaaa
    10681 agcaaaggag cgtttagtga ggggcaattt gactcacctc ccaggcaaca gatgaggggg
    10741 gcaaaaagaa agaaattttc gtgagtcaat atggattccg agcatcattt tcttgcggtc
    10801 tatcttgcta cgtatgttga tcttgacgct gtggatcaag caacgccact cgctcgctcc
    10861 atcgcaggct ggtcgcagac aaattaaaag gcggcaaact cgtacagccg cggggttgtc
    10921 cgctgcaaag tacagagtga taaaagccgc catgcgacca tcaacgcgtt gatgcccagc
    10981 tttttcgatc cgagaatcca ccgtagaggc gatagcaagt aaagaaaagc taaacaaaaa
    11041 aaaatttctg cccctaagcc atgaaaacga gatggggtgg agcagaacca aggaaagagt
    11101 cgcgctgggc tgccgttccg gaaggtgttg taaaggctcg acgcccaagg tgggagtcta
    11161 ggagaagaat ttgcatcggg agtggggcgg gttacccctc catatccaat gacagatatc
    11221 taccagccaa gggtttgagc ccgcccgctt agtcgtcgtc ctcgcttgcc cctccataaa
    11281 aggatttccc ctccccctcc cacaaaattt tctttccctt cctctccttg tccgcttcag
    11341 tacgtatatc ttcccttccc tcgcttctct cctccatcct tctttcatcc atctcctgct
    11401 aacttctctg ctcagcacct ctacgcatta ctagccgtag tatctgagca cttctccctt
    11461 ttatattcca caaaacataa cacaaccttc accatggaca agaagtatag catcgggctg
    11521 gacattggaa cgaactcggt tggttgggct gtgattacgg acgaatacaa ggtgccatcc
    11581 aagaagttta aggtcctggg aaacaccgac cgtcactcaa tcaagaagaa tctcattgga
    11641 gccctgctct tcgatagtgg ggagaccgcc gaagctactc gactgaagcg aacggctcgc
    11701 cggcgttata cacgacgcaa gaatcgcatc tgctacctcc aggagatttt cagcaacgaa
    11761 atggctaagg ttgatgactc attctttcat cgactcgaag aaagtttctt ggtcgaggag
    11821 gataagaagc acgagcgcca tccgatcttt ggtaacattg tggatgaggt tgcctatcac
    11881 gaaaagtacc caactatcta tcatcttcgt aagaagctgg tcgatagcac ggacaaggct
    11941 gatttgcgac ttatctacct ggcactcgcg cacatgatta agttccgcgg ccattttctt
    12001 atcgagggtg acctgaaccc cgataattct gacgttgata agctcttcat ccagttggtc
    12061 caaacctaca atcagctgtt tgaggaaaac cctattaatg catctggcgt ggacgccaag
    12121 gctatccttt cggcgcgcct gtctaagtcg cggcgtttgg agaaccttat cgcacaactc
    12181 cccggcgaaa agaagaacgg cctcttcggt aatttgattg cgttgtcact tggtctgact
    12241 cctaacttca agagtaattt tgacctggca gaggatgcga agctccagtt gtctaaggat
    12301 acgtatgatg acgatctcga caacttgctt gcccaaatcg gtgaccagta cgctgatctt
    12361 ttcctggccg ctaagaatct ctcagatgca atcctgctca gtgacatttt gcgggtcaac
    12421 accgagatta ctaaggcccc cctgtcagct agtatgatca agcggtatga tgagcaccat
    12481 caggacctca ccttgcttaa ggccctcgtg cgtcagcaat tgcctgagaa gtataaggaa
    12541 atcttctttg accaatccaa gaacggatac gcagggtata ttgatggcgg tgcgagccag
    12601 gaggaattct acaagtttat caagccgatt ttggagaaga tggacggcac tgaggaactg
    12661 ctcgtcaagc tgaatcgcga agatttgctt cgtaagcaac gaacgttcga caacggctcc
    12721 atcccgcacc agattcatct gggcgagctc cacgccatcc ttcgacgcca ggaagatttc
    12781 tacccatttc tgaaggacaa ccgtgagaag atcgaaaaga ttcttacatt ccgaatcccc
    12841 tactatgtgg gacctttggc ccgtgggaat tcccgatttg cttggatgac ccgaaagagc
    12901 gaggaaacca tcactccgtg gaacttcgag gaagtcgtgg acaagggtgc atccgcgcag
    12961 agcttcattg agcggatgac caattttgat aagaaccttc cgaatgaaaa ggtcctgcca
    13021 aagcattcgc tgctctacga gtatttcacc gtgtataacg aactgactaa ggtcaagtac
    13081 gtgacggagg gaatgcggaa gccagccttc ctctcagggg aacaaaagaa ggctatcgtc
    13141 gatttgcttt ttaagaccaa tcgtaaagtg actgttaagc agctgaagga ggattatttc
    13201 aagaagattg aatgtttcga ctccgtcgag atcagcggcg tggaagatcg ctttaacgct
    13261 tccctcggta cctaccacga cctgctcaag atcattaagg acaaggattt cctcgataac
    13321 gaggaaaatg aggacatctt ggaagatatt gtcctcacgt tgacactttt tgaggaccgc
    13381 gaaatgatcg aggaacggct caagacatat gcccatttgt tcgacgataa ggtgatgaag
    13441 cagctgaagc ggcgtcgata caccggatgg ggtcgcctta gccggaagct gatcaacggc
    13501 attcgagata agcaatctgg taagactatc ttggatttcc ttaagtcgga cggcttcgcc
    13561 aaccgcaatt ttatgcagct tattcacgac gattccctga cgttcaagga ggacatccag
    13621 aaggcacaag tctcaggaca aggggattcc ctgcacgagc atatcgccaa cctggctgga
    13681 tccccggcga tcaagaaggg gattcttcag accgtcaagg ttgtcgacga gctggtcaag
    13741 gtgatgggcc gtcataagcc agaaaacatc gtgattgaga tggcccgaga aaatcagacc
    13801 actcaaaagg gtcagaagaa cagccgcgag cggatgaagc ggatcgagga aggcattaag
    13861 gaacttggtt ctcagatcct gaaggagcac cctgttgaaa acacacagct ccaaaatgag
    13921 aagctgtatc tctactattt gcaaaatgga cgcgacatgt acgtcgatca ggagctcgac
    13981 attaaccggt tgtcggacta cgatgttgac catatcgtcc cgcaatcctt ccttaaggac
    14041 gatagcattg ataacaaggt gctgactcgc tcagataaga accggggcaa gtccgacaat
    14101 gttccaagcg aggaagtggt taagaagatg aagaactact ggcgccaatt gcttaatgcc
    14161 aagctcatca cacagcgcaa gtttgacaac ttgaccaagg ccgagcgggg agggctgagt
    14221 gaactcgata aggctggctt catcaagcgt caactcgtgg agacgcgaca gatcacaaag
    14281 cacgttgctc agattctgga ctcccggatg aacacaaagt acgacgagaa tgataagctc
    14341 atccgtgaag ttaaggtcat taccctcaag tctaagttgg tgtcggattt ccgcaaggac
    14401 ttccaatttt ataaggttcg ggagatcaac aattatcacc atgcacatga tgcgtacctc
    14461 aacgcagtcg tgggaactgc gctcatcaag aagtatccca agttggagtc cgaattcgtc
    14521 tacggggatt ataaggttta cgacgtccgc aagatgatcg ccaagagtga gcaggaaatt
    14581 ggcaaggcca cggctaagta tttcttttac tccaacatca tgaatttctt taagacggag
    14641 atcacactcg ccaatggaga aatccgtaag cgacctttga ttgagaccaa cggcgagact
    14701 ggtgaaatcg tttgggataa ggggcgcgac ttcgctaccg tgcggaaggt tctgagcatg
    14761 ccgcaagtca atatcgtcaa gaaaaccgag gtgcagacag gcggtttctc taaggaatcg
    14821 attcttccaa agcgtaactc tgacaagctg atcgctcgaa agaaggattg ggaccccaag
    14881 aagtatggag ggttcgattc tcctacagtg gcatactcgg ttctcgttgt cgcgaaggtt
    14941 gagaagggaa agtctaagaa gctgaagtcg gtcaaggaac tgctcgggat caccattatg
    15001 gagcgctcca gcttcgaaaa gaatcccatc gactttctcg aggccaaggg ctataaggaa
    15061 gtcaagaagg atcttatcat taagctgcct aagtactctt tgttcgagct tgaaaacggt
    15121 cgaaagcgaa tgctcgcatc ggcaggagag ttgcagaagg ggaatgaatt ggcacttccc
    15181 tcaaagtacg tgaacttcct gtatctcgcg tcccactacg agaagctgaa gggtagccct
    15241 gaggacaacg aacagaagca actttttgtt gagcaacaca agcattatct ggatgagatc
    15301 attgaacaga tttcagagtt cagtaagcgc gtcatcctcg ccgatgctaa tctcgacaag
    15361 gtgttgtcgg cctacaacaa gcaccgtgac aagccgatcc gagagcaggc tgaaaatatc
    15421 attcatctgt tcaccctcac taacttggga gcaccagcag cgttcaagta ttttgatacg
    15481 acaatcgacc gtaagcgata cacgtccaca aaggaggtgc ttgatgcgac cctgattcat
    15541 caatccatca ctgggctcta tgaaacccgt atcgacctta gtcaactggg gggcgaccct
    15601 cccaagaaga agcgcaaggt ctgagcggac attcgattta tgccgttatg acttccttaa
    15661 aaaagccttt acgaatgaaa gaaatggaat tagacttgtt atgtagttga ttctacaatg
    15721 gattatgatt cctgaacttc aaatccgctg ttcattatta atctcagctc ttcccgtaaa
    15781 gccaatgttg aaactattcg taaatgtacc tcgttttgcg tgtaccttgc ttatcacgtg
    15841 atattacatg acctggacag agttctgcgc gaaagtcata acgtaaatcc cgggcggtag
    15901 gtgcgtcccg ggcggaaggt agttttctcg tccaccccaa cgcgtttatc aacctcaact
    15961 ttcaacaacc atcatgccac caaaagcgcg taaaacaaag cgagatttga ttgagcaaga
    16021 gggcaggatc caatgcgcga ttcaagacat taaaaatgga aaatttcaaa aaattgcgcc
    16081 cgcagcgcgt gcatacaaaa ttcatcccaa tacaagtgta atgctagtgg aggtcaacac
    16141 atcaatgcta ttttggttta gtcgtccagg cggatcacaa aatttgtgtc gtttgacaag
    16201 atggttcatt taggcaactg gtcagatcag cccacttgta agcagtagcg gcggcgctcg
    16261 aagtgtgact cttattagca gacaggaacg aggacattat tatcatctgc tgcttggtgc
    16321 acgataactt gtgcgtttgt caagcaaggt aagtgaacga cccggtcata ccttcttaag
    16381 ttcgcccttc ctccctttat ttcagattca atctgactta cctattctac ccaagcatcc
    16441 aaatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    16501 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    16561 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    16621 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    16681 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    16741 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    16801 ccgaggagca ggactgaccg acgccgacca acaccgccgg tccgacggcg gcccacgggt
    16861 cccaggagct tgagatccac ttaacgttac tgaaatcatc aaacagcttg acgaatctgg
    16921 atataagatc gttggtgtcg atgtcagctc cggagttgag acaaatggtg ttcaggatct
    16981 cgataagata cgttcatttg tccaagcagc aaagagtgcc ttctagtgat ttaatagctc
    17041 catgtcaaca agaataaaac gcgttttcgg gtttacctct tccagataca gctcatctgc
    17101 aatgcattaa tgcattgact gcaacctagt aacgccttnc aggctccggc gaagagaaga
    17161 atagcttagc agagctattt tcattttcgg gagacgagat caagcagatc aacggtcgtc
    17221 aagagaccta cgagactgag gaatccgctc ttggctccac gcgactatat atttgtctct
    17281 aattgtactt tgacatgctc ctcttcttta ctctgatagc ttgactatga aaattccgtc
    17341 accagcncct gggttcgcaa agataattgc atgtttcttc cttgaactct caagcctaca
    17401 ggacacacat tcatcgtagg tataaacctc gaaatcantt cctactaaga tggtatacaa
    17461 tagtaaccat gcatggttgc ctagtgaatg ctccgtaaca cccaatacgc cggccgaaac
    17521 ttttttacaa ctctcctatg agtcgtttac ccagaatgca caggtacact tgtttag
//
