LOCUS       sequence-174613-       17327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AMA1 plasmid with Aspergillus optimized Cas9, argB selection marker 
            and yA specific sgRNA expressed with ribozymes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17327)
  AUTHORS   Nodvig CS, Nielsen JB, Kogle ME, Mortensen UH
  TITLE     A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi.
  JOURNAL   PLoS One. 2015 Jul 15;10(7):e0133085. doi: 
            10.1371/journal.pone.0133085. eCollection 2015.
  PUBMED    26177455
REFERENCE   2  (bases 1 to 17327)
  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: "PLoS One.";
            date: "2015-07-15"; volume: "10(7):e0133085. doi"; pages: " 
            10.1371/journal.pone.0133085. eCollection 2015"
COMMENT     Sequence Label: pFC334
FEATURES             Location/Qualifiers
     source          1..17327
                     /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)"
     intron          7484..7599
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     ncRNA           7615..7651
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
     misc_RNA        7672..7747
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     ncRNA           7752..7819
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     CDS             8955..13058
                     /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             8955..13058
                     /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             13062..13082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(15827..15860)
                     /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(16330..16346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16445..16462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16616..17204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16696..16715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(17326..17327,1..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 gtcaaagact 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 tcacagaaac 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 atgtaaggga
     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 tctgtgagtc
     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 ctttgactag 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 gaagatctca
     7081 tggtcatagc tgtttccgct gagggtttaa tgcgtaagct ccctaattgg cccatccggc
     7141 atctgtaggg cgtccaaata tcgtgcctct cctgctttgc ccggtgtatg aaaccggaaa
     7201 ggccgctcag gagctggcca gcggcgcaga ccgggaacac aagctggcag tcgacccatc
     7261 cggtgctcta cactcgacct gctgaggtcc ctcagtccct ggtaggcagc tttgccccgt
     7321 ctgtccgccc ggtgtgtcgg cggggttgac aaggtcgttg cgtcagtcca acatttgttg
     7381 ccatattttc ctgctctccc caccagctgc tcttttcttt tctctttctt ttcccatctt
     7441 cagtatattc atcttcccat ccaagaacct ttatttcccc taagtaagta ctttgctaca
     7501 tccatactcc atccttccca tcccttattc ctttgaacct ttcagttcga gctttcccac
     7561 ttcatcgcag cttgactaac agctaccccg cttgagcaga catcaccgtc cgccctgatg
     7621 agtccgtgag gacgaaacga gtaagctcgt cggcggagta tcataacatc ggttttagag
     7681 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     7741 tcggtgcttt tggccggcat ggtcccagcc tcctcgctgg cgccggctgg gcaacatgct
     7801 tcggcatggc gaatgggact gatttaatag ctccatgtca acaagaataa aacgcgtttc
     7861 gggtttacct cttccagata cagctcatct gcaatgcatt aatgcattgg acctcgcaac
     7921 cctagtacgc ccttcaggct ccggcgaagc agaagaatag cttagcagag tctattttca
     7981 ttttcgggag acgagatcaa gcagatcaac ggtcgtcaag agacctacga gactgaggaa
     8041 tccgctcttg gctcattaag acctcagccg agacagcaga atcaccgccc aagttaagcc
     8101 tttgtgctga tcatgctctc gaacgggcca agttcgggaa aagcaaagga gcgtttagtg
     8161 aggggcaatt tgactcacct cccaggcaac agatgagggg ggcaaaaaga aagaaatttt
     8221 cgtgagtcaa tatggattcc gagcatcatt ttcttgcggt ctatcttgct acgtatgttg
     8281 atcttgacgc tgtggatcaa gcaacgccac tcgctcgctc catcgcaggc tggtcgcaga
     8341 caaattaaaa ggcggcaaac tcgtacagcc gcggggttgt ccgctgcaaa gtacagagtg
     8401 ataaaagccg ccatgcgacc atcaacgcgt tgatgcccag ctttttcgat ccgagaatcc
     8461 accgtagagg cgatagcaag taaagaaaag ctaaacaaaa aaaaatttct gcccctaagc
     8521 catgaaaacg agatggggtg gagcagaacc aaggaaagag tcgcgctggg ctgccgttcc
     8581 ggaaggtgtt gtaaaggctc gacgcccaag gtgggagtct aggagaagaa tttgcatcgg
     8641 gagtggggcg ggttacccct ccatatccaa tgacagatat ctaccagcca agggtttgag
     8701 cccgcccgct tagtcgtcgt cctcgcttgc ccctccataa aaggatttcc cctccccctc
     8761 ccacaaaatt ttctttccct tcctctcctt gtccgcttca gtacgtatat cttcccttcc
     8821 ctcgcttctc tcctccatcc ttctttcatc catctcctgc taacttctct gctcagcacc
     8881 tctacgcatt actagccgta gtatctgagc acttctccct tttatattcc acaaaacata
     8941 acacaacctt caccatggac aagaagtata gcatcgggct ggacattgga acgaactcgg
     9001 ttggttgggc tgtgattacg gacgaataca aggtgccatc caagaagttt aaggtcctgg
     9061 gaaacaccga ccgtcactca atcaagaaga atctcattgg agccctgctc ttcgatagtg
     9121 gggagaccgc cgaagctact cgactgaagc gaacggctcg ccggcgttat acacgacgca
     9181 agaatcgcat ctgctacctc caggagattt tcagcaacga aatggctaag gttgatgact
     9241 cattctttca tcgactcgaa gaaagtttct tggtcgagga ggataagaag cacgagcgcc
     9301 atccgatctt tggtaacatt gtggatgagg ttgcctatca cgaaaagtac ccaactatct
     9361 atcatcttcg taagaagctg gtcgatagca cggacaaggc tgatttgcga cttatctacc
     9421 tggcactcgc gcacatgatt aagttccgcg gccattttct tatcgagggt gacctgaacc
     9481 ccgataattc tgacgttgat aagctcttca tccagttggt ccaaacctac aatcagctgt
     9541 ttgaggaaaa ccctattaat gcatctggcg tggacgccaa ggctatcctt tcggcgcgcc
     9601 tgtctaagtc gcggcgtttg gagaacctta tcgcacaact ccccggcgaa aagaagaacg
     9661 gcctcttcgg taatttgatt gcgttgtcac ttggtctgac tcctaacttc aagagtaatt
     9721 ttgacctggc agaggatgcg aagctccagt tgtctaagga tacgtatgat gacgatctcg
     9781 acaacttgct tgcccaaatc ggtgaccagt acgctgatct tttcctggcc gctaagaatc
     9841 tctcagatgc aatcctgctc agtgacattt tgcgggtcaa caccgagatt actaaggccc
     9901 ccctgtcagc tagtatgatc aagcggtatg atgagcacca tcaggacctc accttgctta
     9961 aggccctcgt gcgtcagcaa ttgcctgaga agtataagga aatcttcttt gaccaatcca
    10021 agaacggata cgcagggtat attgatggcg gtgcgagcca ggaggaattc tacaagttta
    10081 tcaagccgat tttggagaag atggacggca ctgaggaact gctcgtcaag ctgaatcgcg
    10141 aagatttgct tcgtaagcaa cgaacgttcg acaacggctc catcccgcac cagattcatc
    10201 tgggcgagct ccacgccatc cttcgacgcc aggaagattt ctacccattt ctgaaggaca
    10261 accgtgagaa gatcgaaaag attcttacat tccgaatccc ctactatgtg ggacctttgg
    10321 cccgtgggaa ttcccgattt gcttggatga cccgaaagag cgaggaaacc atcactccgt
    10381 ggaacttcga ggaagtcgtg gacaagggtg catccgcgca gagcttcatt gagcggatga
    10441 ccaattttga taagaacctt ccgaatgaaa aggtcctgcc aaagcattcg ctgctctacg
    10501 agtatttcac cgtgtataac gaactgacta aggtcaagta cgtgacggag ggaatgcgga
    10561 agccagcctt cctctcaggg gaacaaaaga aggctatcgt cgatttgctt tttaagacca
    10621 atcgtaaagt gactgttaag cagctgaagg aggattattt caagaagatt gaatgtttcg
    10681 actccgtcga gatcagcggc gtggaagatc gctttaacgc ttccctcggt acctaccacg
    10741 acctgctcaa gatcattaag gacaaggatt tcctcgataa cgaggaaaat gaggacatct
    10801 tggaagatat tgtcctcacg ttgacacttt ttgaggaccg cgaaatgatc gaggaacggc
    10861 tcaagacata tgcccatttg ttcgacgata aggtgatgaa gcagctgaag cggcgtcgat
    10921 acaccggatg gggtcgcctt agccggaagc tgatcaacgg cattcgagat aagcaatctg
    10981 gtaagactat cttggatttc cttaagtcgg acggcttcgc caaccgcaat tttatgcagc
    11041 ttattcacga cgattccctg acgttcaagg aggacatcca gaaggcacaa gtctcaggac
    11101 aaggggattc cctgcacgag catatcgcca acctggctgg atccccggcg atcaagaagg
    11161 ggattcttca gaccgtcaag gttgtcgacg agctggtcaa ggtgatgggc cgtcataagc
    11221 cagaaaacat cgtgattgag atggcccgag aaaatcagac cactcaaaag ggtcagaaga
    11281 acagccgcga gcggatgaag cggatcgagg aaggcattaa ggaacttggt tctcagatcc
    11341 tgaaggagca ccctgttgaa aacacacagc tccaaaatga gaagctgtat ctctactatt
    11401 tgcaaaatgg acgcgacatg tacgtcgatc aggagctcga cattaaccgg ttgtcggact
    11461 acgatgttga ccatatcgtc ccgcaatcct tccttaagga cgatagcatt gataacaagg
    11521 tgctgactcg ctcagataag aaccggggca agtccgacaa tgttccaagc gaggaagtgg
    11581 ttaagaagat gaagaactac tggcgccaat tgcttaatgc caagctcatc acacagcgca
    11641 agtttgacaa cttgaccaag gccgagcggg gagggctgag tgaactcgat aaggctggct
    11701 tcatcaagcg tcaactcgtg gagacgcgac agatcacaaa gcacgttgct cagattctgg
    11761 actcccggat gaacacaaag tacgacgaga atgataagct catccgtgaa gttaaggtca
    11821 ttaccctcaa gtctaagttg gtgtcggatt tccgcaagga cttccaattt tataaggttc
    11881 gggagatcaa caattatcac catgcacatg atgcgtacct caacgcagtc gtgggaactg
    11941 cgctcatcaa gaagtatccc aagttggagt ccgaattcgt ctacggggat tataaggttt
    12001 acgacgtccg caagatgatc gccaagagtg agcaggaaat tggcaaggcc acggctaagt
    12061 atttctttta ctccaacatc atgaatttct ttaagacgga gatcacactc gccaatggag
    12121 aaatccgtaa gcgacctttg attgagacca acggcgagac tggtgaaatc gtttgggata
    12181 aggggcgcga cttcgctacc gtgcggaagg ttctgagcat gccgcaagtc aatatcgtca
    12241 agaaaaccga ggtgcagaca ggcggtttct ctaaggaatc gattcttcca aagcgtaact
    12301 ctgacaagct gatcgctcga aagaaggatt gggaccccaa gaagtatgga gggttcgatt
    12361 ctcctacagt ggcatactcg gttctcgttg tcgcgaaggt tgagaaggga aagtctaaga
    12421 agctgaagtc ggtcaaggaa ctgctcggga tcaccattat ggagcgctcc agcttcgaaa
    12481 agaatcccat cgactttctc gaggccaagg gctataagga agtcaagaag gatcttatca
    12541 ttaagctgcc taagtactct ttgttcgagc ttgaaaacgg tcgaaagcga atgctcgcat
    12601 cggcaggaga gttgcagaag gggaatgaat tggcacttcc ctcaaagtac gtgaacttcc
    12661 tgtatctcgc gtcccactac gagaagctga agggtagccc tgaggacaac gaacagaagc
    12721 aactttttgt tgagcaacac aagcattatc tggatgagat cattgaacag atttcagagt
    12781 tcagtaagcg cgtcatcctc gccgatgcta atctcgacaa ggtgttgtcg gcctacaaca
    12841 agcaccgtga caagccgatc cgagagcagg ctgaaaatat cattcatctg ttcaccctca
    12901 ctaacttggg agcaccagca gcgttcaagt attttgatac gacaatcgac cgtaagcgat
    12961 acacgtccac aaaggaggtg cttgatgcga ccctgattca tcaatccatc actgggctct
    13021 atgaaacccg tatcgacctt agtcaactgg ggggcgaccc tcccaagaag aagcgcaagg
    13081 tctgagcgga cattcgattt atgccgttat gacttcctta aaaaagcctt tacgaatgaa
    13141 agaaatggaa ttagacttgt tatgtagttg attctacaat ggattatgat tcctgaactt
    13201 caaatccgct gttcattatt aatctcagct cttcccgtaa agccaatgtt gaaactattc
    13261 gtaaatgtac ctcgttttgc gtgtaccttg cttatcacgt gatattacat gacctggaca
    13321 gagttctgcg cgaaagtcat aacgtaaatc ccgggcggta ggtgcgtccc gggcggaagg
    13381 tagttttctc gtccacccca acgcgtttat caacctcaac tttcaacaac catcatgcca
    13441 ccaaaagcgc gtaaaacaaa gcgagatttg attgagcaag agggcaggat ccaatgcgcg
    13501 attcaagaca ttaaaaatgg aaaatttcaa aaaattgcgc ccgcagcgcg tgcatacaaa
    13561 attcatccca atacaagtgt aatactaggt aatatcgcgt gcattccgcg gttttttggg
    13621 gtagtcatct aatgaaacag acccggacgc agcagaggaa gcccgcgatg actctatacc
    13681 accgtacgcc gatatatcat catcgcggcg atggagaagt tgggttgact ccgaagacac
    13741 ttcaaaggag cgacgctgtt gatttgtaga cgacgcttga tagggagaag cattattgtc
    13801 gtgatgctcg cccaacagag gccgactcgc ctcatccgtc ataacgaacg ctgtgtaaag
    13861 cggagtgggg gggaaagtgt ggattgtgga gagtatgcga tagtgttgag gctgatcaga
    13921 cggcgaatcg ggccagatat gaccagttta gaggcctcat ttgactataa tttacataaa
    13981 ttagataaat agagatgaac gcatgcaata attgcagcaa atattgatga agcgagaggt
    14041 aggacgatga aggactgtga gcagttcaag gtatcagcag agtcaagggc ctgatgcaat
    14101 ggcggtgatc cgtgatcagc gaacggaagg ggcgctaact ctgtttcttt accaatgatc
    14161 ggaagctcct gctggcggac ttatgagtca ttcacgaatc atttctcagt tatttgtgga
    14221 tgccctcgtt ctgtccacaa tttctttccg ccccaagtct tttaagttct ttaacatcta
    14281 tattcttgca cttccaatgg catcccttcg ctccgtactc aagagccaga gcttgcgaca
    14341 caccgtgcga tcctactcct cgcaaaccat gcctcccgcc tcaccctttg ctccccgcca
    14401 cttcctctcc attgcggacc tctcgccctc cgagttcgca acccttgttc gcaatgcctc
    14461 ctcacacaaa cgggctatca agtcggggtc aatgccccaa aacttgcagg gatcactcct
    14521 tgggaaaact gtggccatga tcttcagcaa acgaagcacg aggacaaggg tatctacaga
    14581 aggggccgtt gtgcagatgg gaggtcatcc gatgttcttg ggcaaggatg atatccaact
    14641 aggtgtcaac gagtccctat acgacacctc cgttgtcatt tcgtccatgg tatcctgcat
    14701 tgtagcccgt gtcggtaaac atgcagaggt cgcagatctg gcgaagcact cttcggttcc
    14761 agtcatcaat gctttgtgtg actctttcca ccctctccaa gccgtggccg atttccagac
    14821 catctatgaa gcattcaccc ccaaggcgca ccacctttca agtctagggt tggaaggatt
    14881 gaagatcgct tgggtgggtg acgccaacaa cgtcctgttc gatatggcca ttgctgctac
    14941 aaaaatgggt gtcgacattg ctgtcgctac tcccaagggg tacgaaatcc ctcctcacat
    15001 gctggagctc atcaagtctg ctggagaggg tgtctcgaaa ccaggaaagc ttctgcaaac
    15061 caatattccc gaagaagcgg tcaaggacgc cgatattctg gtcacagaca cctgggtctc
    15121 tatgggccaa gaggaagaga aggctcagag gctgaaggag tttgatggtt tccaaatcac
    15181 tgctgaactc gccaagcgag gaggagctaa ggagggctgg aagttcatgc actgtctccc
    15241 gcgacaccct gaggaggtca gcgacgaggt tttctacagc aaccggtcac ttgtcttccc
    15301 tgaggctgag aaccggttat gggctgcgat ttccgccttg gagggtttcg ttgtcaataa
    15361 gggaaaaatt gaataaatat aaccaggctt ccatttaaat atatagaggc tggcgttatc
    15421 aaactgatga gttgacgggt atgagatcat tccgtcccta aatatattta ctccgatcac
    15481 gtaaaagcct gttagtagaa gcattttccc aattatcctg accaattctt tctagcatat
    15541 atcaaataac taattgacat gttctctcgc ttccttatat tactcagagt attggaaatg
    15601 gggcaaatcg cacccggtga ctttcacatg tcacgaatgc ggagtcgtcc tagccaaggt
    15661 agatccaggc ctaacacacc ccaaccctcg actctcctca ttccatactc ttgacctcta
    15721 tccagcacat tcttctgagg tttcgcaatg gctgtaggtg gcgctgaggc atccccacta
    15781 ccgcattaag acctcagcgc ggccgcaaat ttaaataaaa tgaagtgaag ttcctatact
    15841 ttctagagaa taggaacttc tatagtgagt cgaataaggg cgacacaaaa tttattctaa
    15901 atgcataata aatactgata acatcttata gtttgtatta tattttgtat tatcgttgac
    15961 atgtataatt ttgatatcaa aaactgattt tccctttatt attttcgaga tttattttct
    16021 taattctctt taacaaacta gaaatattgt atatacaaaa aatcataaat aatagatgaa
    16081 tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt atcttattta aagtgcgttg
    16141 cttttttctc atttataagg ttaaataatt ctcatatatc aagcaaagtg acaggcgccc
    16201 ttaaatattc tgacaaatgc tctttcccta aactcccccc ataaaaaaac ccgccgaagc
    16261 gggtttttac gttatttgcg gattaacgat tactcgttat cagaaccgcc cagggggccc
    16321 gagcttaaga ctggccgtcg ttttacaaca cagaaagagt ttgtagaaac gcaaaaaggc
    16381 catccgtcag gggccttctg cttagtttga tgcctggcag ttccctactc tcgccttccg
    16441 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
    16501 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
    16561 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
    16621 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
    16681 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
    16741 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    16801 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    16861 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    16921 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    16981 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tgggctaact
    17041 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    17101 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    17161 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    17221 tttctacggg gtctgacgct cagtggaacg acgcgcgcgt aactcacgtt aagggatttt
    17281 ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc tctgctt
//
