LOCUS       sequence-430137-       17590 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 17590)
  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 17590)
  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..17590
                     /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(1562..2353,2354..2422))
                     /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     2185..2204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2423..2514)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2556..2575
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2581..2600
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    2807..2840
                     /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      3153..8405
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     ncRNA           9114..9181
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     RBS             9341..9352
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             9359..10072
                     /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      10084..10155
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      10171..10198
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        10216..10291
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             11497..15600
                     /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             11497..15600
                     /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             15604..15624
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             16450..16824
                     /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 atcaaaaaag 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 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1621 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1681 ccagcgctgc gatgataccg cgagaaccac gctcaccggc tccggattta tcagcaataa
     1741 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1801 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1861 acgttgttgc catcgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1921 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     1981 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2041 tcatggttat ggcagcgcta cataattctc ttactgtcat gccatccgta agatgctttt
     2101 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2161 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2221 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2281 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2341 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2401 cacggaaatg ttgaatactc atattcttcc tttttcaata ttattgaagc atttatcagg
     2461 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2521 tcagtgttac aaccaattaa ccaattctga acattatcgc gagcccattt atacctgaat
     2581 atggctcata acaccccttg tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     2641 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggac tccccatgcg
     2701 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2761 tcgcccgggc taattatggg gtgtcgccct tattcgactc tatagtgaag ttcctattct
     2821 ctagaaagta taggaacttc tgaagtgggg atttaaatgc ggccgcgctg agggtttaat
     2881 cgacgaagca gctgacggcc agtgccaagc ttaacgcgta ccgggcccag tatatgttcc
     2941 gcagatgact ggagctctgc catacgtgcc ctctcaagca ccatttgttc catctacaga
     3001 gactagtcac caactagtct atcaagactc acagggtaca ttgctgagac caactgacca
     3061 gaggcagggt agcggattga cggctccatc tccttcactt acaaggtcta ttgaaagccc
     3121 tttagcatca ccaagcggag aatagattgt taagcttatt ttttgtatac tgttttgtga
     3181 tagcacgaag tttttccacg gtatcttgtt aaaaatatat atttgtggcg ggcttaccta
     3241 catcaaatta ataagagact aattataaac taaacacaca agcaagctac tttagggtaa
     3301 aagtttataa atgcttttga cgtataaacg ttgcttgtat ttattattac aattaaaggt
     3361 ggatagaaaa cctagagact agttagaaac taatctcagg tttgcgttaa actaaatcag
     3421 agcccgagag gttaacagaa cctagaaggg gactagatat ccgggtaggg aaacaaaaaa
     3481 aaaaaannca agacagccac atattaggga gactagttag aagctagttc caggactagg
     3541 aaaataaaag acaatgatac cacagtctag ttgacaacta gatagattct agattgaggc
     3601 caaagtctct gagatccagg ttagttgcaa ctaatactag ttagtatcta gtctcctata
     3661 actctgaagc tagaataact tactactatt atcctcacca ctgttcagct gcgcaaacgg
     3721 agtgattgca aggtgttcag agactagtta ttgactagtc agtgactagc aataactaac
     3781 aaggtattaa cctaccatgt ctgccatcac cctgcacttc ctcgggctca gcagcctttt
     3841 cctcctcatt ttcatgctca ttttccttgt ttaagactgt gactagtcaa agactagtcc
     3901 agaaccacaa aggagaaatg tcttaccact ttcttcattg cttgtctctt ttgcattatc
     3961 catgtctgca actagttaga gtctagttag tgactagtcc gacgaggact tgcttgtctc
     4021 cggattgttg gaggaactct ccagggcctc aagatccaca acagagcctt ctagaagact
     4081 ggtcaataac tagttggtct ttgtctgagt ctgacttacg aggttgcata ctcgctccct
     4141 ttgcctcgtc aatcgatgag aaaaagcgcc aaaactcgca atatggcttt gaaccacacg
     4201 gtgctgagac tagttagaat ctagtcccaa actagcttgg atagcttacc tttgcccttt
     4261 gcgttgcgac aggtcttgca gggtatggtt cctttctcac cagctgattt agctgccttg
     4321 ctaccctcac ggcggatctg cataaagagt ggctagaggt tataaattag cactgatcct
     4381 aggtacgggg ctgaatgtaa cttgccyttc ctttctcatc gcgcggcaag acaggcttgc
     4441 tcaaattcct accagtcaca ggggtatgca cggcgtacgg accacttgaa ctagtcacag
     4501 attagttagc aactagtctg cattgaatgg ctgtacttac gggccctcgc cattgtcctg
     4561 atcatttcca gcttcaccct cgttgctgca aagtagttag tgactagtca aggactagtt
     4621 gaaatgggag aagaaactca cgaattctcg acacccttag tattgtggtc cttggacttg
     4681 gtgctgctat atattagcta atacactagt tagactcaca gaaacttacg cagctcgctt
     4741 gcgcttcttg gtaggagtcg gggttgggag aacagtgcct tcaaacaagc cttcatacca
     4801 tgctacttga ctagtcaggg actagtcacc aagtaatcta gataggactt gcctttggcc
     4861 tccatcagtt ccttcatagt gggaggtcca ttgtgcaatg taaactccat gccgtgggag
     4921 ttcttgtcct tcaagtgctt gaccaatatg tttctgttgg cagagggaac ctgtcaacta
     4981 gttaataact agtcagaaac tagtatagca gtagactcac tgtacgcttg aggcatccct
     5041 tcactcggca gtagacttca tatggatgga tatcaggcac gccattgtcg tcctgtggac
     5101 tagtcagtaa ctaggcttaa agctagtcgg gtcggcttac tatcttgaaa tccggcagcg
     5161 taagctcccc gtccttaact gcctcgagat agtgacagta ctctggggac tttcggagat
     5221 cgttatcgcg aatgctcggc atactaatcg ttgactagtc ttggactagt cccgagcaaa
     5281 aaggattgga ggaggaggag gaaggtgaga gtgagacaaa gagcgaaata agagcttcaa
     5341 aggctatctc taagcagtat gaaggttaag tatctagttc ttgactagat ttaaaagaga
     5401 tttcgactag ttatgtacct ggagtttgga tataggaatg tgttgtggta acgaaatgta
     5461 agggggagga aagaaaaagt cggtcaagag gtaactctaa gtcggccatt cctttttggg
     5521 aggcgctaac cataaacggc atggtcgact tagagttagc tcagggaatt tagggagtta
     5581 tctgcgacca ccgaggaacg gcggaatgcc aaagaatccc gatggagctc tagctggcgg
     5641 ttgacaaccc caccttttgg cgtttctgcg gcgttgcagg cgggactgga tacttcgtag
     5701 aaccagaaag gcaaggcaga acgcgctcag caagagtgtt ggaagtgata gcatgatgtg
     5761 ccttgttaac taggtcaaaa tctgcagtat gcttgatgtt atccgaagtg tgagagagga
     5821 aggtccaaac atacacgatt gggagagggc ctaggtataa gagtttttga gtagaacgca
     5881 tgtgagccca gccatctcga ggagattaaa cacgggccgg catttgatgg ctatgttagt
     5941 accccaatgg aaacggtgag agtccagtgg tcgcagataa ctccctaaat tccctgagct
     6001 aactctaagt cgaccatgcc gtttatggtt agcgcctccc aaaaaggaat ggccgactta
     6061 gagttacctc ttgaccgact ttttctttcc tcccccttac atttcgttac cacaacacat
     6121 tcctatatcc aaactccagg tacataacta gtcgaaatct cttttaaatc tagtcaagaa
     6181 ctagatactt aaccttcata ctgcttagag atagcctttg aagctcttat ttcgctcttt
     6241 gtctcactct caccttcctc ctcctcctcc aatccttttt gctcgggact agtccaagac
     6301 tagtcaacga ttagtatgcc gagcattcgc gataacgatc tccgaaagtc cccagagtac
     6361 tgtcactatc tcgaggcagt taaggacggg gagcttacgc tgccggattt caagatagta
     6421 agccgacccg actagcttta agcctagtta ctgactagtc cacaggacga caatggcgtg
     6481 cctgatatcc atccatatga agtctactgc cgagtgaagg gatgcctcaa gcgtacagtg
     6541 agtctactgc tatactagtt tctgactagt tattaactag ttgacaggtt ccctctgcca
     6601 acagaaacat attggtcaag cacttgaagg acaagaactc ccacggcatg gagtttacat
     6661 tgcacaatgg acctcccact atgaaggaac tgatggaggc caaaggcaag tcctatctag
     6721 attacttggt gactagtccc tgactagtca agtagcatgg tatgaaggct tgtttgaagg
     6781 cactgttctc ccaaccccga ctcctaccaa gaagcgcaag cgagctgcgt aagtttctgt
     6841 gagtctaact agtgtattag ctaatatata gcagcaccaa gtccaaggac cacaatacta
     6901 agggtgtcga gaattcgtga gtttcttctc ccatttcaac tagtccttga ctagtcacta
     6961 actactttgc agcaacgagg gtgaagctgg aaatgatcag gacaatggcg agggcccgta
     7021 agtacagcca ttcaatgcag actagttgct aactaatctg tgactagttc aagtggtccg
     7081 tacgccgtgc atacccctgt gactggtagg aatttgagca agcctgtctt gccgcgcgat
     7141 gagaaaggaa rggcaagtta cattcagccc cgtacctagg atcagtgcta atttataacc
     7201 tctagccact ctttatgcag atccgccgtg agggtagcaa ggcagctaaa tcagctggtg
     7261 agaaaggaac cataccctgc aagacctgtc gcaacgcaaa gggcaaaggt aagctatcca
     7321 agctagtttg ggactagatt ctaactagtc tcagcaccgt gtggttcaaa gccatattgc
     7381 gagttttggc gctttttctc atcgattgac gaggcaaagg gagcgagtat gcaacctcgt
     7441 aagtcagact cagacaaaga ccaactagtt attgaccagt cttctagaag gctctgttgt
     7501 ggatcttgag gccctggaga gttcctccaa caatccggag acaagcaagt cctcgtcgga
     7561 ctagtcacta actagactct aactagttgc agacatggat aatgcaaaag agacaagcaa
     7621 tgaagaaagt ggtaagacat ttctcctttg tggttctgga ctagtctttg actagtcaca
     7681 gtcttaaaca aggaaaatga gcatgaaaat gaggaggaaa aggctgctga gcccgaggaa
     7741 gtgcagggtg atggcagaca tggtaggtta ataccttgtt agttattgct agtcactgac
     7801 tagtcaataa ctagtctctg aacaccttgc aatcactccg tttgcgcagc tgaacagtgg
     7861 tgaggataat agtagtaagt tattctagct tcagagttat aggagactag atactaacta
     7921 gtattagttg caactaacct ggatctcaga gactttggcc tcaatctaga atctatctag
     7981 ttgtcaacta gactgtggta tcattgtctt ttattttcct agtcctggaa ctagcttcta
     8041 actagtctcc ctaatatgtg gctgtcttgt tttttttttt tttgtttccc tacccggata
     8101 tctagtcccc ttctaggttc tgttaacctc tcgggctctg atttagttta acgcaaacct
     8161 gagattagtt tctaactagt ctctaggttt tctatccacc tttaattgta ataataaata
     8221 caagcaacgt ttatacgtca aaagcattta taaactttta ccctaaagta gcttgcttgt
     8281 gtgtttagtt tataattagt ctcttattaa tttgatgtag gtaagcccgc cacaaatata
     8341 tatttttaac aagataccgt ggaaaaactt cgtgctatca caaaacagta tacaaaaaat
     8401 aagcttaaca atctattctc cgcttggtga tgctaaaggg ctttcaatag accttgtaag
     8461 tgaaggagat ggagccgtca atccgctacc ctgcctctgg tcagttggtc tcagcaatgt
     8521 accctgtgag tcttgataga ctagttggtg actagtctct gtagatggaa caaatggtgc
     8581 ttgagagggc acgtatggca gagctccagt catctgcgga acatatactg ggcccgggaa
     8641 gatctgatca catagatgct cggttgacag gaccacctat agcaagtact ttgtaggagg
     8701 ttctcctaac gcttggcttc aaatccaatc aggatatcag tgagttactg tttgtcatcg
     8761 catcctcatg aagtcgctca acagccactg agaagcaaat atttgggaga ccatccctga
     8821 tgttgaaatt ttacgctgga gcccattcac cggtgagctc ggggacagtc tggccagtgg
     8881 agcggaaaat cttcaactaa ctctgattgg taccacaggt agccccacag aacaaaccga
     8941 gcaaacatga aaattttcgc ttgaggttag cgcactcgct agcgcctgcc cgcaaatata
     9001 aaaggccccg aaatcccatg cattttggaa atcaagcgac tctacgtatg tcatccccct
     9061 ccctaccttt gggaaaacct ctcaactgca agtcagaaca ttttgctaac agcggccggc
     9121 atggtcccag cctcctcgct ggcgccggct gggcaacatg cttcggcatg gcgaatggga
     9181 ccgatgcagg tggaaagtga aacgtgattt catgcgtcat tttgaacatt ttgtaaatct
     9241 tatttaataa tgtgtgcggc aattcacatt taatttatga atgttttctt aacatcgcgg
     9301 caactcaaga aacggcaggt tcggatctta gctactagag aaagaggaga aatactagat
     9361 gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga
     9421 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa
     9481 actgacgctg aagttcatct gtactactgg taaactgccg gttccttggc cgactctggt
     9541 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca
     9601 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa
     9661 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa
     9721 ccgcattgag ctgaaaggca ttgactttaa agaggacggc aatatcctgg gccataagct
     9781 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat
     9841 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca
     9901 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct
     9961 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct
    10021 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgaccagg
    10081 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
    10141 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
    10201 tatacacctg cagtcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
    10261 tcaacttgaa aaagtggcac cgagtcggtg cttttgcatc attggtctag tggtagaatt
    10321 catcgttgcc atcgatgagg cccgtgttcg attcacggat gatgcatgtc aatttttaca
    10381 cttgatgttg gtgtaatcag gcgatcagag tttgctcaaa ttctctcttg ttttagattc
    10441 tgaagaatat aatttttaca tgactagttc attttacttc caagtagact atttgtagat
    10501 tgagattgcc gtcaacttta ttcatagaat cacattaaat tgctcattga cctgcatgtc
    10561 tatgcacaca ttcacatcta tcttctcagg gtattaatta agacctcagc cgagacagca
    10621 gaatcaccgc ccaagttaag cctttgtgct gatcatgctc tcgaacgggc caagttcggg
    10681 aaaagcaaag gagcgtttag tgaggggcaa tttgactcac ctcccaggca acagatgagg
    10741 ggggcaaaaa gaaagaaatt ttcgtgagtc aatatggatt ccgagcatca ttttcttgcg
    10801 gtctatcttg ctacgtatgt tgatcttgac gctgtggatc aagcaacgcc actcgctcgc
    10861 tccatcgcag gctggtcgca gacaaattaa aaggcggcaa actcgtacag ccgcggggtt
    10921 gtccgctgca aagtacagag tgataaaagc cgccatgcga ccatcaacgc gttgatgccc
    10981 agctttttcg atccgagaat ccaccgtaga ggcgatagca agtaaagaaa agctaaacaa
    11041 aaaaaaattt ctgcccctaa gccatgaaaa cgagatgggg tggagcagaa ccaaggaaag
    11101 agtcgcgctg ggctgccgtt ccggaaggtg ttgtaaaggc tcgacgccca aggtgggagt
    11161 ctaggagaag aatttgcatc gggagtgggg cgggttaccc ctccatatcc aatgacagat
    11221 atctaccagc caagggtttg agcccgcccg cttagtcgtc gtcctcgctt gcccctccat
    11281 aaaaggattt cccctccccc tcccacaaaa ttttctttcc cttcctctcc ttgtccgctt
    11341 cagtacgtat atcttccctt ccctcgcttc tctcctccat ccttctttca tccatctcct
    11401 gctaacttct ctgctcagca cctctacgca ttactagccg tagtatctga gcacttctcc
    11461 cttttatatt ccacaaaaca taacacaacc ttcaccatgg acaagaagta tagcatcggg
    11521 ctggacattg gaacgaactc ggttggttgg gctgtgatta cggacgaata caaggtgcca
    11581 tccaagaagt ttaaggtcct gggaaacacc gaccgtcact caatcaagaa gaatctcatt
    11641 ggagccctgc tcttcgatag tggggagacc gccgaagcta ctcgactgaa gcgaacggct
    11701 cgccggcgtt atacacgacg caagaatcgc atctgctacc tccaggagat tttcagcaac
    11761 gaaatggcta aggttgatga ctcattcttt catcgactcg aagaaagttt cttggtcgag
    11821 gaggataaga agcacgagcg ccatccgatc tttggtaaca ttgtggatga ggttgcctat
    11881 cacgaaaagt acccaactat ctatcatctt cgtaagaagc tggtcgatag cacggacaag
    11941 gctgatttgc gacttatcta cctggcactc gcgcacatga ttaagttccg cggccatttt
    12001 cttatcgagg gtgacctgaa ccccgataat tctgacgttg ataagctctt catccagttg
    12061 gtccaaacct acaatcagct gtttgaggaa aaccctatta atgcatctgg cgtggacgcc
    12121 aaggctatcc tttcggcgcg cctgtctaag tcgcggcgtt tggagaacct tatcgcacaa
    12181 ctccccggcg aaaagaagaa cggcctcttc ggtaatttga ttgcgttgtc acttggtctg
    12241 actcctaact tcaagagtaa ttttgacctg gcagaggatg cgaagctcca gttgtctaag
    12301 gatacgtatg atgacgatct cgacaacttg cttgcccaaa tcggtgacca gtacgctgat
    12361 cttttcctgg ccgctaagaa tctctcagat gcaatcctgc tcagtgacat tttgcgggtc
    12421 aacaccgaga ttactaaggc ccccctgtca gctagtatga tcaagcggta tgatgagcac
    12481 catcaggacc tcaccttgct taaggccctc gtgcgtcagc aattgcctga gaagtataag
    12541 gaaatcttct ttgaccaatc caagaacgga tacgcagggt atattgatgg cggtgcgagc
    12601 caggaggaat tctacaagtt tatcaagccg attttggaga agatggacgg cactgaggaa
    12661 ctgctcgtca agctgaatcg cgaagatttg cttcgtaagc aacgaacgtt cgacaacggc
    12721 tccatcccgc accagattca tctgggcgag ctccacgcca tccttcgacg ccaggaagat
    12781 ttctacccat ttctgaagga caaccgtgag aagatcgaaa agattcttac attccgaatc
    12841 ccctactatg tgggaccttt ggcccgtggg aattcccgat ttgcttggat gacccgaaag
    12901 agcgaggaaa ccatcactcc gtggaacttc gaggaagtcg tggacaaggg tgcatccgcg
    12961 cagagcttca ttgagcggat gaccaatttt gataagaacc ttccgaatga aaaggtcctg
    13021 ccaaagcatt cgctgctcta cgagtatttc accgtgtata acgaactgac taaggtcaag
    13081 tacgtgacgg agggaatgcg gaagccagcc ttcctctcag gggaacaaaa gaaggctatc
    13141 gtcgatttgc tttttaagac caatcgtaaa gtgactgtta agcagctgaa ggaggattat
    13201 ttcaagaaga ttgaatgttt cgactccgtc gagatcagcg gcgtggaaga tcgctttaac
    13261 gcttccctcg gtacctacca cgacctgctc aagatcatta aggacaagga tttcctcgat
    13321 aacgaggaaa atgaggacat cttggaagat attgtcctca cgttgacact ttttgaggac
    13381 cgcgaaatga tcgaggaacg gctcaagaca tatgcccatt tgttcgacga taaggtgatg
    13441 aagcagctga agcggcgtcg atacaccgga tggggtcgcc ttagccggaa gctgatcaac
    13501 ggcattcgag ataagcaatc tggtaagact atcttggatt tccttaagtc ggacggcttc
    13561 gccaaccgca attttatgca gcttattcac gacgattccc tgacgttcaa ggaggacatc
    13621 cagaaggcac aagtctcagg acaaggggat tccctgcacg agcatatcgc caacctggct
    13681 ggatccccgg cgatcaagaa ggggattctt cagaccgtca aggttgtcga cgagctggtc
    13741 aaggtgatgg gccgtcataa gccagaaaac atcgtgattg agatggcccg agaaaatcag
    13801 accactcaaa agggtcagaa gaacagccgc gagcggatga agcggatcga ggaaggcatt
    13861 aaggaacttg gttctcagat cctgaaggag caccctgttg aaaacacaca gctccaaaat
    13921 gagaagctgt atctctacta tttgcaaaat ggacgcgaca tgtacgtcga tcaggagctc
    13981 gacattaacc ggttgtcgga ctacgatgtt gaccatatcg tcccgcaatc cttccttaag
    14041 gacgatagca ttgataacaa ggtgctgact cgctcagata agaaccgggg caagtccgac
    14101 aatgttccaa gcgaggaagt ggttaagaag atgaagaact actggcgcca attgcttaat
    14161 gccaagctca tcacacagcg caagtttgac aacttgacca aggccgagcg gggagggctg
    14221 agtgaactcg ataaggctgg cttcatcaag cgtcaactcg tggagacgcg acagatcaca
    14281 aagcacgttg ctcagattct ggactcccgg atgaacacaa agtacgacga gaatgataag
    14341 ctcatccgtg aagttaaggt cattaccctc aagtctaagt tggtgtcgga tttccgcaag
    14401 gacttccaat tttataaggt tcgggagatc aacaattatc accatgcaca tgatgcgtac
    14461 ctcaacgcag tcgtgggaac tgcgctcatc aagaagtatc ccaagttgga gtccgaattc
    14521 gtctacgggg attataaggt ttacgacgtc cgcaagatga tcgccaagag tgagcaggaa
    14581 attggcaagg ccacggctaa gtatttcttt tactccaaca tcatgaattt ctttaagacg
    14641 gagatcacac tcgccaatgg agaaatccgt aagcgacctt tgattgagac caacggcgag
    14701 actggtgaaa tcgtttggga taaggggcgc gacttcgcta ccgtgcggaa ggttctgagc
    14761 atgccgcaag tcaatatcgt caagaaaacc gaggtgcaga caggcggttt ctctaaggaa
    14821 tcgattcttc caaagcgtaa ctctgacaag ctgatcgctc gaaagaagga ttgggacccc
    14881 aagaagtatg gagggttcga ttctcctaca gtggcatact cggttctcgt tgtcgcgaag
    14941 gttgagaagg gaaagtctaa gaagctgaag tcggtcaagg aactgctcgg gatcaccatt
    15001 atggagcgct ccagcttcga aaagaatccc atcgactttc tcgaggccaa gggctataag
    15061 gaagtcaaga aggatcttat cattaagctg cctaagtact ctttgttcga gcttgaaaac
    15121 ggtcgaaagc gaatgctcgc atcggcagga gagttgcaga aggggaatga attggcactt
    15181 ccctcaaagt acgtgaactt cctgtatctc gcgtcccact acgagaagct gaagggtagc
    15241 cctgaggaca acgaacagaa gcaacttttt gttgagcaac acaagcatta tctggatgag
    15301 atcattgaac agatttcaga gttcagtaag cgcgtcatcc tcgccgatgc taatctcgac
    15361 aaggtgttgt cggcctacaa caagcaccgt gacaagccga tccgagagca ggctgaaaat
    15421 atcattcatc tgttcaccct cactaacttg ggagcaccag cagcgttcaa gtattttgat
    15481 acgacaatcg accgtaagcg atacacgtcc acaaaggagg tgcttgatgc gaccctgatt
    15541 catcaatcca tcactgggct ctatgaaacc cgtatcgacc ttagtcaact ggggggcgac
    15601 cctcccaaga agaagcgcaa ggtctgagcg gacattcgat ttatgccgtt atgacttcct
    15661 taaaaaagcc tttacgaatg aaagaaatgg aattagactt gttatgtagt tgattctaca
    15721 atggattatg attcctgaac ttcaaatccg ctgttcatta ttaatctcag ctcttcccgt
    15781 aaagccaatg ttgaaactat tcgtaaatgt acctcgtttt gcgtgtacct tgcttatcac
    15841 gtgatattac atgacctgga cagagttctg cgcgaaagtc ataacgtaaa tcccgggcgg
    15901 taggtgcgtc ccgggcggaa ggtagttttc tcgtccaccc caacgcgttt atcaacctca
    15961 actttcaaca accatcatgc caccaaaagc gcgtaaaaca aagcgagatt tgattgagca
    16021 agagggcagg atccaatgcg cgattcaaga cattaaaaat ggaaaatttc aaaaaattgc
    16081 gcccgcagcg cgtgcataca aaattcatcc caatacaagt gtaatgctag tggaggtcaa
    16141 cacatcaatg cctattttgg tttagtcgtc caggcggtga gcacaaaatt tgtgtcgttt
    16201 gacaagatgg ttcatttagg caactggtca gatcagcccc acttgtagca gtagcggcgg
    16261 cgctcgaagt gtgactctta ttagcagaca ggaacgagga cattattatc atctgctgct
    16321 tggtgcacga taacttggtg cgtttgtcaa gcaaggtaag tggacgaccc ggtcatacct
    16381 tcttaagttc gcccttcctc cctttatttc agattcaatc tgacttacct attctaccca
    16441 agcatccaaa tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
    16501 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
    16561 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
    16621 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
    16681 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
    16741 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
    16801 ttcgtggccg aggagcagga ctgaccgacg ccgaccaaca ccgccggtcc gacgcggccc
    16861 gacgggtccg aggagcttga gatccactta acgttactga aatcatcaaa cagcttgacg
    16921 aatctggata taagatcgtt ggtgtcgatg tcagctccgg agttgagaca aatggtgttc
    16981 aggatctcga taagatacgt tcatttgtcc aagcagcaaa gagtgccttc tagtgattta
    17041 atagctccat gtcaacaaga ataaaacgcg tttcgggttt acctcttcca gatacagctc
    17101 atctgcaatg cattaatgca ttggacctcg caaccctagt acgcccttca ggctccggcg
    17161 aagcagaaga atagcttagc agagtctatt ttcattttcg ggagacgaga tcaagcagat
    17221 caacggtcgt caagagacct acgagactga ggaatccgct cttggctcca cgcgactata
    17281 tatttgtctc taattgtact ttgacatgct cctcttcttt actctgatag cttgactatg
    17341 aaaattccgt caccagcccc tgggttcgca aagataattg cactgtttct tccttgaact
    17401 ctcaagccta caggacacac attcatcgta ggtataaacc tcgaaaatca ttcctactaa
    17461 gatgggtata caatagtaac catgcatggt tgcctagtga atgctccgta acacccaata
    17521 cgccggccga aactttttta caactctcct atgagtcgtt tacccagaat gcacaggtac
    17581 acttgtttag
//
