LOCUS       sequence-429385-       17673 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 pyrG 
            selectable marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17673)
  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 17673)
  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: pDM026
FEATURES             Location/Qualifiers
     source          1..17673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /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      1590..6838
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     ncRNA           7547..7614
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     RBS             7774..7785
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7792..8505
                     /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
                     NFKIRHNVEDGSVQLAYHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      8517..8588
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8604..8631
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        8649..8724
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             9930..14033
                     /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             9930..14033
                     /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             14037..14057
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(16173..16206)
                     /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(16676..16692)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16791..16808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16962..17550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(17042..17061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(17672..17673,1..790,791..859))
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcgctacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatt taaatgcggc cgcgctgagg gtttaatcga
     1321 cgaagcagct gacggccagt gccaagctta acgcgtaccg ggcccagtat atgttccgca
     1381 gatgactgga gctctgccat acgtgccctc tcaagcacca tttgttccat ctacagagac
     1441 tagtcaccaa ctagtctatc aagactcaca gggtacattg ctgagaccaa ctgaccagag
     1501 gcagggtagc ggattgacgg ctccatctcc ttcacttaca aggtctattg aaagcccttt
     1561 agcatcacca agcggagaat agattgttaa gcttattttt tgtatactgt tttgtgatag
     1621 cacgaagttt ttccacggta tcttgttaaa aatatatatt tgtggcgggc ttacctacat
     1681 caaattaata agagactaat tataaactaa acacacaagc aagctacttt agggtaaaag
     1741 tttataaatg cttttgacgt ataaacgttg cttgtattta ttattacaat taaaggtgga
     1801 tagaaaacct agagactagt tagaaactaa tctcaggttt gcgttaaact aaatcagagc
     1861 ccgagaggtt aacagaacct agaaggggac tagatatccg ggtagggaaa caaaaaaaaa
     1921 aancaagaca gccacatatt agggagacta gttagaagct agttccagga ctaggaaaat
     1981 aaaagacaat gataccacag tctagttgac aactagatag attctagatt gaggccaaag
     2041 tctctgagat ccaggttagt tgcaactaat actagttagt atctagtctc ctataactct
     2101 gaagctagaa taacttacta ctattatcct caccactgtt cagctgcgca aacggagtga
     2161 ttgcaaggtg ttcagagact agttattgac tagtcagtga ctagcaataa ctaacaaggt
     2221 attaacctac catgtctgcc atcaccctgc acttcctcgg gctcagcagc cttttcctcc
     2281 tcattttcat gctcattttc cttgtttaag actgtgacta gtcraagact agtccagaac
     2341 cacaaaggag aaatgtctta ccactttctt cattgcttgt ctcttttgca ttatccatgt
     2401 ctgcaactag ttagagtcta gttagtgact agtccgacga ggacttgctt gtctccggat
     2461 tgttggagga actctccagg gcctcaagat ccacaacaga gccttctaga agactggtca
     2521 ataactagtt ggtctttgtc tgagtctgac ttacgaggtt gcatactcgc tccctttgcc
     2581 tcgtcaatcg atgagaaaaa gcgccaaaac tcgcaatatg gctttgaacc acacggtgct
     2641 gagactagtt agaatctagt cccaaactag cttggatagc ttacctttgc cctttgcgtt
     2701 gcgacaggtc ttgcagggta tggttccttt ctcaccagct gatttagctg ccttgctacc
     2761 ctcacggcgg atctgcataa agagtggcta gaggttataa attagcactg atcctaggta
     2821 cggggctgaa tgtaacttgc cyttcctttc tcatcgcgcg gcaagacagg cttgctcaaa
     2881 ttcctaccag tcacaggggt atgcacggcg tacggaccac ttgaactagt cacagattag
     2941 ttagcaacta gtctgcattg aatggctgta cttacgggcc ctcgccattg tcctgatcat
     3001 ttccagcttc accctcgttg ctgcaaagta gttagtgact agtcaaggac tagttgaaat
     3061 gggagaagaa actcacgaat tctcgacacc cttagtattg tggtccttgg acttggtgct
     3121 gctatatatt agctaataca ctagttagac tcacrgaaac ttacgcagct cgcttgcgct
     3181 tcttggtagg agtcggggtt gggagaacag tgccttcaaa caagccttca taccatgcta
     3241 cttgactagt cagggactag tcaccaagta atctagatag gacttgcctt tggcctccat
     3301 cagttccttc atagtgggag gtccattgtg caatgtaaac tccatgccgt gggagttctt
     3361 gtccttcaag tgcttgacca atatgtttct gttggcagag ggaacctgtc aactagttaa
     3421 taactagtca gaaactagta tagcagtaga ctcactgtac gcttgaggca tcccttcact
     3481 cggcagtaga cttcatatgg atggatatca ggcacgccat tgtcgtcctg tggactagtc
     3541 agtaactagg cttaaagcta gtcgggtcgg cttactatct tgaaatccgg cagcgtaagc
     3601 tccccgtcct taactgcctc gagatagtga cagtactctg gggactttcg gagatcgtta
     3661 tcgcgaatgc tcggcatact aatcgttgac tagtcttgga ctagtcccga gcaaaaagga
     3721 ttggaggagg aggaggaagg tgagagtgag acaaagagcg aaataagagc ttcaaaggct
     3781 atctctaagc agtatgaagg ttaagtatct agttcttgac tagatttaaa agagatttcg
     3841 actagttatg tacctggagt ttggatatag gaatgtgttg tggtaacgaa atgtaagggg
     3901 gaggaaagaa aaagtcggtc aagaggtaac tctaagtcgg ccattccttt ttgggaggcg
     3961 ctaaccataa acggcatggt cgacttagag ttagctcagg gaatttaggg agttatctgc
     4021 gaccaccgag gaacggcgga atgccaaaga atcccgatgg agctctagct ggcggttgac
     4081 aaccccacct tttggcgttt ctgcggcgtt gcaggcggga ctggatactt cgtagaacca
     4141 gaaaggcaag gcagaacgcg ctcagcaaga gtgttggaag tgatagcatg atgtgccttg
     4201 ttaactaggt caaaatctgc agtatgcttg atgttatcca aagtgtgaga gaggaaggtc
     4261 caaacataca cgattgggag agggcctagg tataagagtt tttgagtaga acgcatgtga
     4321 gcccagccat ctcgaggaga ttaaacacgg gccggcattt gatggctatg ttagtacccc
     4381 aatggaaacg gtgagagtcc agtggtcgca gataactccc taaattccct gagctaactc
     4441 taagtcgacc atgccgttta tggttagcgc ctcccaaaaa ggaatggccg acttagagtt
     4501 acctcttgac cgactttttc tttcctcccc cttacatttc gttaccacaa cacattccta
     4561 tatccaaact ccaggtacat aactagtcga aatctctttt aaatctagtc aagaactaga
     4621 tacttaacct tcatactgct tagagatagc ctttgaagct cttatttcgc tctttgtctc
     4681 actctcacct tcctcctcct cctccaatcc tttttgctcg ggactagtcc aagactagtc
     4741 aacgattagt atgccgagca ttcgcgataa cgatctccga aagtccccag agtactgtca
     4801 ctatctcgag gcagttaagg acggggagct tacgctgccg gatttcaaga tagtaagccg
     4861 acccgactag ctttaagcct agttactgac tagtccacag gacgacaatg gcgtgcctga
     4921 tatccatcca tatgaagtct actgccgagt gaagggatgc ctcaagcgta cagtgagtct
     4981 actgctatac tagtttctga ctagttatta actagttgac aggttccctc tgccaacaga
     5041 aacatattgg tcaagcactt gaaggacaag aactcccacg gcatggagtt tacattgcac
     5101 aatggacctc ccactatgaa ggaactgatg gaggccaaag gcaagtccta tctagattac
     5161 ttggtgacta gtccctgact agtcaagtag catggtatga aggcttgttt gaaggcactg
     5221 ttctcccaac cccgactcct accaagaagc gcaagcgagc tgcgtaagtt tcygtgagtc
     5281 taactagtgt attagctaat atatagcagc accaagtcca aggaccacaa tactaagggt
     5341 gtcgagaatt cgtgagtttc ttctcccatt tcaactagtc cttgactagt cactaactac
     5401 tttgcagcaa cgagggtgaa gctggaaatg atcaggacaa tggcgagggc ccgtaagtac
     5461 agccattcaa tgcagactag ttgctaacta atctgtgact agttcaagtg gtccgtacgc
     5521 cgtgcatacc cctgtgactg gtaggaattt gagcaagcct gtcttgccgc gcgatgagaa
     5581 aggaarggca agttacattc agccccgtac ctaggatcag tgctaattta taacctctag
     5641 ccactcttta tgcagatccg ccgtgagggt agcaaggcag ctaaatcagc tggtgagaaa
     5701 ggaaccatac cctgcaagac ctgtcgcaac gcaaagggca aaggtaagct atccaagcta
     5761 gtttgggact agattctaac tagtctcagc accgtgtggt tcaaagccat attgcgagtt
     5821 ttggcgcttt ttctcatcga ttgacgaggc aaagggagcg agtatgcaac ctcgtaagtc
     5881 agactcagac aaagaccaac tagttattga ccagtcttct agaaggctct gttgtggatc
     5941 ttgaggccct ggagagttcc tccaacaatc cggagacaag caagtcctcg tcggactagt
     6001 cactaactag actctaacta gttgcagaca tggataatgc aaaagagaca agcaatgaag
     6061 aaagtggtaa gacatttctc ctttgtggtt ctggactagt cttygactag tcacagtctt
     6121 aaacaaggaa aatgagcatg aaaatgagga ggaaaaggct gctgagcccg aggaagtgca
     6181 gggtgatggc agacatggta ggttaatacc ttgttagtta ttgctagtca ctgactagtc
     6241 aataactagt ctctgaacac cttgcaatca ctccgtttgc gcagctgaac agtggtgagg
     6301 ataatagtag taagttattc tagcttcaga gttataggag actagatact aactagtatt
     6361 agttgcaact aacctggatc tcagagactt tggcctcaat ctagaatcta tctagttgtc
     6421 aactagactg tggtatcatt gtcttttatt ttcctagtcc tggaactagc ttctaactag
     6481 tctccctaat atgtggctgt cttgtttttt ttttttgttt ccctacccgg atatctagtc
     6541 cccttctagg ttctgttaac ctctcgggct ctgatttagt ttaacgcaaa cctgagatta
     6601 gtttctaact agtctctagg ttttctatcc acctttaatt gtaataataa atacaagcaa
     6661 cgtttatacg tcaaaagcat ttataaactt ttaccctaaa gtagcttgct tgtgtgttta
     6721 gtttataatt agtctcttat taatttgatg taggtaagcc cgccacaaat atatattttt
     6781 aacaagatac cgtggaaaaa cttcgtgcta tcacaaaaca gtatacaaaa aataagctta
     6841 acaatctatt ctccgcttgg tgatgctaaa gggctttcaa tagaccttgt aagtgaagga
     6901 gatggagccg tcaatccgct accctgcctc tggtcagttg gtctcagcaa tgtaccctgt
     6961 gagtcttgat agactagttg gtgactagtc tctgtagatg gaacaaatgg tgcttgagag
     7021 ggcacgtatg gcagagctcc agtcatctgc ggaacatata ctgggcccgg gaagatctga
     7081 tcacatagat gctcggttga caggaccacc tatagcaagt actttgtagg aggttctcct
     7141 aacgcttggc ttcaaatcca atcaggatat cagtgagtta ctgtttgtca tcgcatcctc
     7201 atgaagtcgc tcaacagcca ctgagaagca aatatttggg agaccatccc tgatgttgaa
     7261 attttacgct ggagcccatt caccggtgag ctcggggaca gtctggccag tggagcggaa
     7321 aatcttcaac taactctgat tggtaccaca ggtagcccca cagaacaaac cgagcaaaca
     7381 tgaaaatttt cgcttgaggt tagcgcactc gctagcgcct gcccgcaaat ataaaaggcc
     7441 ccgaaatccc atgcattttg gaaatcaagc gactctacgt atgtcatccc cctccctacc
     7501 tttgggaaaa cctctcaact gcaagtcaga acattttgct aacagcggcc ggcatggtcc
     7561 cagcctcctc gctggcgccg gctgggcaac atgcttcggc atggcgaatg ggaccgatgc
     7621 aggtggaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa tcttatttaa
     7681 taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg cggcaactca
     7741 agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta gatgcgtaaa
     7801 ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac
     7861 ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg
     7921 ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct ggtaacgacg
     7981 ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc
     8041 ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac
     8101 ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt
     8161 gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa gctggaatac
     8221 aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg
     8281 aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctta tcactaccag
     8341 caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg
     8401 caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc
     8461 gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc aggcatcaaa
     8521 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     8581 acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatacac
     8641 ctgcagtcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     8701 gaaaaagtgg caccgagtcg gtgcttttgc atcattggtc tagtggtaga attcatcgtt
     8761 gccatcgatg aggcccgtgt tcgattcacg gatgatgcat gtcaattttt acacttgatg
     8821 ttggtgtaat caggcgatca gagtttgctc aaattctctc ttgttttaga ttctgaagaa
     8881 tataattttt acatgactag ttcattttac ttccaagtag actatttgta gattgagatt
     8941 gccgtcaact ttattcatag aatcacatta aattgctcat tgacctgcat gtctatgcac
     9001 acattcacat ctatcttctc agggtattaa ttaagacctc agccgagaca gcagaatcac
     9061 cgcccaagtt aagcctttgt gctgatcatg ctctcgaacg ggccaagttc gggaaaagca
     9121 aaggagcgtt tagtgagggg caatttgact cacctcccag gcaacagatg aggggggcaa
     9181 aaagaaagaa attttcgtga gtcaatatgg attccgagca tcattttctt gcggtctatc
     9241 ttgctacgta tgttgatctt gacgctgtgg atcaagcaac gccactcgct cgctccatcg
     9301 caggctggtc gcagacaaat taaaaggcgg caaactcgta cagccgcggg gttgtccgct
     9361 gcaaagtaca gagtgataaa agccgccatg cgaccatcaa cgcgttgatg cccagctttt
     9421 tcgatccgag aatccaccgt agaggcgata gcaagtaaag aaaagctaaa caaaaaaaaa
     9481 tttctgcccc taagccatga aaacgagatg gggtggagca gaaccaagga aagagtcgcg
     9541 ctgggctgcc gttccggaag gtgttgtaaa ggctcgacgc ccaaggtggg agtctaggag
     9601 aagaatttgc atcgggagtg gggcgggtta cccctccata tccaatgaca gatatctacc
     9661 agccaagggt ttgagcccgc ccgcttagtc gtcgtcctcg cttgcccctc cataaaagga
     9721 tttcccctcc ccctcccaca aaattttctt tcccttcctc tccttgtccg cttcagtacg
     9781 tatatcttcc cttccctcgc ttctctcctc catccttctt tcatccatct cctgctaact
     9841 tctctgctca gcacctctac gcattactag ccgtagtatc tgagcacttc tcccttttat
     9901 attccacaaa acataacaca accttcacca tggacaagaa gtatagcatc gggctggaca
     9961 ttggaacgaa ctcggttggt tgggctgtga ttacggacga atacaaggtg ccatccaaga
    10021 agtttaaggt cctgggaaac accgaccgtc actcaatcaa gaagaatctc attggagccc
    10081 tgctcttcga tagtggggag accgccgaag ctactcgact gaagcgaacg gctcgccggc
    10141 gttatacacg acgcaagaat cgcatctgct acctccagga gattttcagc aacgaaatgg
    10201 ctaaggttga tgactcattc tttcatcgac tcgaagaaag tttcttggtc gaggaggata
    10261 agaagcacga gcgccatccg atctttggta acattgtgga tgaggttgcc tatcacgaaa
    10321 agtacccaac tatctatcat cttcgtaaga agctggtcga tagcacggac aaggctgatt
    10381 tgcgacttat ctacctggca ctcgcgcaca tgattaagtt ccgcggccat tttcttatcg
    10441 agggtgacct gaaccccgat aattctgacg ttgataagct cttcatccag ttggtccaaa
    10501 cctacaatca gctgtttgag gaaaacccta ttaatgcatc tggcgtggac gccaaggcta
    10561 tcctttcggc gcgcctgtct aagtcgcggc gtttggagaa ccttatcgca caactccccg
    10621 gcgaaaagaa gaacggcctc ttcggtaatt tgattgcgtt gtcacttggt ctgactccta
    10681 acttcaagag taattttgac ctggcagagg atgcgaagct ccagttgtct aaggatacgt
    10741 atgatgacga tctcgacaac ttgcttgccc aaatcggtga ccagtacgct gatcttttcc
    10801 tggccgctaa gaatctctca gatgcaatcc tgctcagtga cattttgcgg gtcaacaccg
    10861 agattactaa ggcccccctg tcagctagta tgatcaagcg gtatgatgag caccatcagg
    10921 acctcacctt gcttaaggcc ctcgtgcgtc agcaattgcc tgagaagtat aaggaaatct
    10981 tctttgacca atccaagaac ggatacgcag ggtatattga tggcggtgcg agccaggagg
    11041 aattctacaa gtttatcaag ccgattttgg agaagatgga cggcactgag gaactgctcg
    11101 tcaagctgaa tcgcgaagat ttgcttcgta agcaacgaac gttcgacaac ggctccatcc
    11161 cgcaccagat tcatctgggc gagctccacg ccatccttcg acgccaggaa gatttctacc
    11221 catttctgaa ggacaaccgt gagaagatcg aaaagattct tacattccga atcccctact
    11281 atgtgggacc tttggcccgt gggaattccc gatttgcttg gatgacccga aagagcgagg
    11341 aaaccatcac tccgtggaac ttcgaggaag tcgtggacaa gggtgcatcc gcgcagagct
    11401 tcattgagcg gatgaccaat tttgataaga accttccgaa tgaaaaggtc ctgccaaagc
    11461 attcgctgct ctacgagtat ttcaccgtgt ataacgaact gactaaggtc aagtacgtga
    11521 cggagggaat gcggaagcca gccttcctct caggggaaca aaagaaggct atcgtcgatt
    11581 tgctttttaa gaccaatcgt aaagtgactg ttaagcagct gaaggaggat tatttcaaga
    11641 agattgaatg tttcgactcc gtcgagatca gcggcgtgga agatcgcttt aacgcttccc
    11701 tcggtaccta ccacgacctg ctcaagatca ttaaggacaa ggatttcctc gataacgagg
    11761 aaaatgagga catcttggaa gatattgtcc tcacgttgac actttttgag gaccgcgaaa
    11821 tgatcgagga acggctcaag acatatgccc atttgttcga cgataaggtg atgaagcagc
    11881 tgaagcggcg tcgatacacc ggatggggtc gccttagccg gaagctgatc aacggcattc
    11941 gagataagca atctggtaag actatcttgg atttccttaa gtcggacggc ttcgccaacc
    12001 gcaattttat gcagcttatt cacgacgatt ccctgacgtt caaggaggac atccagaagg
    12061 cacaagtctc aggacaaggg gattccctgc acgagcatat cgccaacctg gctggatccc
    12121 cggcgatcaa gaaggggatt cttcagaccg tcaaggttgt cgacgagctg gtcaaggtga
    12181 tgggccgtca taagccagaa aacatcgtga ttgagatggc ccgagaaaat cagaccactc
    12241 aaaagggtca gaagaacagc cgcgagcgga tgaagcggat cgaggaaggc attaaggaac
    12301 ttggttctca gatcctgaag gagcaccctg ttgaaaacac acagctccaa aatgagaagc
    12361 tgtatctcta ctatttgcaa aatggacgcg acatgtacgt cgatcaggag ctcgacatta
    12421 accggttgtc ggactacgat gttgaccata tcgtcccgca atccttcctt aaggacgata
    12481 gcattgataa caaggtgctg actcgctcag ataagaaccg gggcaagtcc gacaatgttc
    12541 caagcgagga agtggttaag aagatgaaga actactggcg ccaattgctt aatgccaagc
    12601 tcatcacaca gcgcaagttt gacaacttga ccaaggccga gcggggaggg ctgagtgaac
    12661 tcgataaggc tggcttcatc aagcgtcaac tcgtggagac gcgacagatc acaaagcacg
    12721 ttgctcagat tctggactcc cggatgaaca caaagtacga cgagaatgat aagctcatcc
    12781 gtgaagttaa ggtcattacc ctcaagtcta agttggtgtc ggatttccgc aaggacttcc
    12841 aattttataa ggttcgggag atcaacaatt atcaccatgc acatgatgcg tacctcaacg
    12901 cagtcgtggg aactgcgctc atcaagaagt atcccaagtt ggagtccgaa ttcgtctacg
    12961 gggattataa ggtttacgac gtccgcaaga tgatcgccaa gagtgagcag gaaattggca
    13021 aggccacggc taagtatttc ttttactcca acatcatgaa tttctttaag acggagatca
    13081 cactcgccaa tggagaaatc cgtaagcgac ctttgattga gaccaacggc gagactggtg
    13141 aaatcgtttg ggataagggg cgcgacttcg ctaccgtgcg gaaggttctg agcatgccgc
    13201 aagtcaatat cgtcaagaaa accgaggtgc agacaggcgg tttctctaag gaatcgattc
    13261 ttccaaagcg taactctgac aagctgatcg ctcgaaagaa ggattgggac cccaagaagt
    13321 atggagggtt cgattctcct acagtggcat actcggttct cgttgtcgcg aaggttgaga
    13381 agggaaagtc taagaagctg aagtcggtca aggaactgct cgggatcacc attatggagc
    13441 gctccagctt cgaaaagaat cccatcgact ttctcgaggc caagggctat aaggaagtca
    13501 agaaggatct tatcattaag ctgcctaagt actctttgtt cgagcttgaa aacggtcgaa
    13561 agcgaatgct cgcatcggca ggagagttgc agaaggggaa tgaattggca cttccctcaa
    13621 agtacgtgaa cttcctgtat ctcgcgtccc actacgagaa gctgaagggt agccctgagg
    13681 acaacgaaca gaagcaactt tttgttgagc aacacaagca ttatctggat gagatcattg
    13741 aacagatttc agagttcagt aagcgcgtca tcctcgccga tgctaatctc gacaaggtgt
    13801 tgtcggccta caacaagcac cgtgacaagc cgatccgaga gcaggctgaa aatatcattc
    13861 atctgttcac cctcactaac ttgggagcac cagcagcgtt caagtatttt gatacgacaa
    13921 tcgaccgtaa gcgatacacg tccacaaagg aggtgcttga tgcgaccctg attcatcaat
    13981 ccatcactgg gctctatgaa acccgtatcg accttagtca actggggggc gaccctccca
    14041 agaagaagcg caaggtctga gcggacattc gatttatgcc gttatgactt ccttaaaaaa
    14101 gcctttacga atgaaagaaa tggaattaga cttgttatgt agttgattct acaatggatt
    14161 atgattcctg aacttcaaat ccgctgttca ttattaatct cagctcttcc cgtaaagcca
    14221 atgttgaaac tattcgtaaa tgtacctcgt tttgcgtgta ccttgcttat cacgtgatat
    14281 tacatgacct ggacagagtt ctgcgcgaaa gtcataacgt aaatcccggg cggtaggtgc
    14341 gtcccgggcg gaaggtagtt ttctcgtcca ccccaacgcg tttatcaacc tcaactttca
    14401 acaaccatca tgccaccaaa agcgcgtaaa acaaagcgag atttgattga gcaagagggc
    14461 aggatccaat gcgcgattca agacattaaa aatggaaaat ttcaaaaaat tgcgcccgca
    14521 gcgcgtgcat acaaaattca tcccaataca agtgtaattc ctcgtgtact gtgtaagcgc
    14581 ccactaggta atatgacatg attacgaatt cgagctcggt acccggccgg ggatcctcta
    14641 gacgtggagt taccagtgat tgaccaatgt tttatcttct acagttctgc ctgtctaccc
    14701 cattctagct gtacctgact acagagtagt ttaattgtgg ttgaccccac agtcggaggc
    14761 ggaggaatac agcaccgatg tggcctgtct ccatccagat tggcacgcaa tttttacacg
    14821 cggaaaagat cgagatagag tacgacttta aatttagtcc ccggcggctt ctattttaga
    14881 atatttgaga tttgattctc aagcaattga tttggttggg tcaccctcaa ttggataata
    14941 tacctcattg ctcggctact tcaactcatc aatcaccgtc ataccccgca tataaccctc
    15001 cattcccacg atgtcgtcca agtcgcaatt gacttacggt gctcgagcca gcaagcaccc
    15061 caatcctctg gcaaagagac tttttgagat tgccgaagca aagaagacaa acgttaccgt
    15121 ctctgctgat gtgacgacaa cccgagaact cctggacctc gctgaccgta cggaagctgt
    15181 tggatccaat acatatgccg tctagcaatg gactaatcaa cttttgatga tacaggtctc
    15241 ggtccctaca tcgccgtcat caagacacac atcgacatcc tcaccgattt cagcgtcgac
    15301 actatcaatg gcctgaatgt gctggctcaa aagtacaact ttttgatctt cgaggaccgc
    15361 aaattcatcg acatcggcaa taccgtccag aagcaatacc acggcggtgc tctgaggatc
    15421 tccgaatggg cccacattat caactgcagc gttctccctg gcgagggcat cgtcgaggct
    15481 ctggcccaga ccgcatctgc gcaagacttc ccctatggtc ctgagagagg actgttggtc
    15541 ctggcagaga tgacccccaa aggatcgctg gctacgggcg agtataccaa ggcatcggtt
    15601 gactacgctc gcaaatacaa gaacttcgtt atgggtttcg tgtcgacgcg ggccctgacg
    15661 gaagtgcagt cggatgtgtc ttcagcctcg gaggatgaag atttcgtggt cttcacgacg
    15721 ggtgtgaacc tctcttccaa aggagataag cttggacagc aataccagac tcctgcatcg
    15781 gctattggac gcggtgccga ctttatcatc gccggtcgag gcatctacgc tgctcccgac
    15841 ccggttgaag ctgcacagcg gtaccagaaa gaaggctggg aagcttatat ggccagagta
    15901 tgcggcaagt catgatttcc tcttggagca aaagtgtagt gccagtacga gtgttgtgga
    15961 ggaaggctgc atacattgtg cctgtcatta aacgatgagc tcgtccgtat tggcccctgt
    16021 aatgccatgt tttccgcccc caatcgtcaa ggttttccct ttgttagatt cctaccagtc
    16081 atctagcaag gcggccgcag ctagcacaat tgaggcatcc ccactaccgc attaagacct
    16141 cagcgcggcc gcaaatttaa ataaaatgaa gtgaagttcc tatactttct agagaatagg
    16201 aacttctata gtgagtcgaa taagggcgac acaaaattta ttctaaatgc ataataaata
    16261 ctgataacat cttatagttt gtattatatt ttgtattatc gttgacatgt ataattttga
    16321 tatcaaaaac tgattttccc tttattattt tcgagattta ttttcttaat tctctttaac
    16381 aaactagaaa tattgtatat acaaaaaatc ataaataata gatgaatagt ttaattatag
    16441 gtgttcatca atcgaaaaag caacgtatct tatttaaagt gcgttgcttt tttctcattt
    16501 ataaggttaa ataattctca tatatcaagc aaagtgacag gcgcccttaa atattctgac
    16561 aaatgctctt tccctaaact ccccccataa aaaaacccgc cgaagcgggt ttttacgtta
    16621 tttgcggatt aacgattact cgttatcaga accgcccagg gggcccgagc ttaagactgg
    16681 ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa aaaggccatc cgtcaggggc
    16741 cttctgctta gtttgatgcc tggcagttcc ctactctcgc cttccgcttc ctcgctcact
    16801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    16861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    16921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    16981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    17041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    17101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    17161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    17221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    17281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    17341 aggtatgtag gcggtgctac agagttcttg aagtggtggg ctaactacgg ctacactaga
    17401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    17461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    17521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    17581 gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg gattttggtc atgagcttgc
    17641 gccgtcccgt caagtcagcg taatgctctg ctt
//
