LOCUS       sequence-251142-       10647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multigene editing in the Fusarium fujikuroi genome using the 
            CRISPR-Cas9 system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10647)
  AUTHORS   Shi TQ, Gao J, Wang WJ, Wang KF, Xu GQ, Huang H, Ji XJ
  TITLE     CRISPR/Cas9-Based Genome Editing in the Filamentous Fungus Fusarium 
            fujikuroi and Its Application in Strain Engineering for Gibberellic 
            Acid Production.
  JOURNAL   ACS Synth Biol. 2019 Feb 15;8(2):445-454. doi: 
            10.1021/acssynbio.8b00478. Epub 2019 Jan 23.
  PUBMED    30616338
REFERENCE   2  (bases 1 to 10647)
  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"; doi: 
            "10.1021/acssynbio.8b00478"; journalName: "ACS Synth Biol"; date: 
            "2019-02-15- 15"; volume: "8"; issue: "2"; pages: "445-454"
COMMENT     Sequence Label: pUC-fFuCas9-HTBNLS-hph
FEATURES             Location/Qualifiers
     source          1..10647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(612..1403,1404..1472))
                     /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     1235..1254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1473..1577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1645..1663
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1701..1723)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1823..1842
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2036..2058
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2050..2067
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2051..2067
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     intron          2907..3022
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             3031..7134
                     /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             3031..7134
                     /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             8488..9513
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(9585..9618)
                     /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(10098..10114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10098..10114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10111..10133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10122..10138
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(10146..10152,10153..10170,10171..10176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10191..10212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10329..10346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(10500..10647,1..441))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10580..10599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
       61 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      121 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      181 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      241 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      301 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      361 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      421 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      481 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
      541 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
      601 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
      661 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
      721 ccatctggcc ccagtgctgc aatgataccg cgactcccac gctcaccggc tccagattta
      781 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
      841 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
      901 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
      961 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1021 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1081 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1141 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1201 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1261 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1321 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     1381 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     1441 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     1501 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     1561 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     1621 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     1681 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     1741 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1801 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccagatg
     1861 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     1921 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     1981 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     2041 tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtacctcgcg aatgcatcta
     2101 gatatcggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgt acagtgaccg
     2161 gtgactcttt ctggcatgcg gagagacgga cggacgcaga gagaagggct gagtaataag
     2221 cgccactgcg ccagacagct ctggcggctc tgaggtgcag tggatgatta ttaatccggg
     2281 accggccgcc cctccgcccc gaagtggaaa ggctggtgtg cccctcgttg accaagaatc
     2341 tattgcatca tcggagaata tggagcttca tcgaatcacc ggcagtaagc gaaggagaat
     2401 gtgaagccag gggtgtatag ccgtcggcga aatagcatgc cattaaccta ggtacagaag
     2461 tccaattgct tccgatctgg taaaagattc acgagatagt accttctccg aagtaggtag
     2521 agcgagtacc cggcgcgtaa gctccctaat tggcccatcc ggcatctgta gggcgtccaa
     2581 atatcgtgcc tctcctgctt tgcccggtgt atgaaaccgg aaaggccgct caggagctgg
     2641 ccagcggcgc agaccgggaa cacaagctgg cagtcgaccc atccggtgct ctgcactcga
     2701 cctgctgagg tccctcagtc cctggtaggc agctttgccc cgtctgtccg cccggtgtgt
     2761 cggcggggtt gacaaggtcg ttgcgtcagt ccaacatttg ttgccatatt ttcctgctct
     2821 ccccaccagc tgctcttttc ttttctcttt cttttcccat cttcagtata ttcatcttcc
     2881 catccaagaa cctttatttc ccctaagtaa gtactttgct acatccatac tccatccttc
     2941 ccatccctta ttcctttgaa cctttcagtt cgagctttcc cacttcatcg cagcttgact
     3001 aacagctacc ccgcttgagc agacatcacc atggacaaga agtactctat cggcctcgac
     3061 atcggtacta actcagttgg ttgggccgtt atcacagacg agtacaaggt tcccagcaag
     3121 aagttcaagg tccttggtaa caccgaccga cactctatca agaagaacct catcggcgct
     3181 ctgcttttcg attcaggaga gacagccgag gctactcgac ttaaacgaac agctcgacga
     3241 cgatacactc gacgaaagaa ccgcatttgc tacctccagg agatcttcag caacgagatg
     3301 gctaaggtcg acgatagctt cttccatcga cttgaggaga gcttcctcgt tgaggaggat
     3361 aagaagcacg agcgacatcc tatcttcggt aacatcgtcg acgaggttgc ttaccacgag
     3421 aagtacccta ccatctacca ccttcgcaag aagctcgtcg attctaccga taaggccgat
     3481 cttcgactca tctacctcgc tcttgctcat atgatcaagt tccgcggtca cttcctcatc
     3541 gagggcgatc ttaacccaga taactcagac gtcgacaagc tcttcatcca gctcgttcaa
     3601 acctacaacc agctcttcga ggagaaccct atcaacgctt caggagttga cgctaaggct
     3661 atcctctcag ctcgactctc taagtctcga cgacttgaga acctcatcgc tcaacttcct
     3721 ggcgagaaaa agaacggtct cttcggtaac cttatcgctc tctctctcgg tcttactcct
     3781 aacttcaaga gcaacttcga ccttgccgag gacgctaagc tccagctctc taaggatacc
     3841 tacgacgacg atctcgataa cctccttgct cagatcggag accaatacgc cgatcttttc
     3901 cttgccgcta agaacctttc agacgctatc ctcctcagcg atatccttcg agtcaacact
     3961 gagatcacta aggctcctct ctcagcttct atgatcaagc gctacgacga gcatcatcag
     4021 gatctcactc tcctcaaggc tcttgttcga caacagcttc ccgagaagta caaggagatc
     4081 ttcttcgacc agtctaagaa cggttacgcc ggttatattg acggaggagc tagccaagag
     4141 gagttctaca agttcatcaa gcccatcctc gagaagatgg acggtacaga ggagcttctc
     4201 gttaagctca accgagagga tctccttcgt aagcagcgaa ctttcgacaa cggtagcatc
     4261 cctcatcaga tccatcttgg agagcttcac gctatccttc gacgacagga ggacttctat
     4321 cctttcctca aggacaaccg cgagaagatc gagaagatcc tcactttccg catcccttac
     4381 tacgttggtc ctcttgctcg aggtaactct cgatttgctt ggatgacccg aaagtcagag
     4441 gagactatca ccccttggaa cttcgaggag gttgttgaca agggagcttc agctcagtct
     4501 ttcatcgagc gcatgaccaa cttcgacaag aacctcccta acgagaaggt cctccctaag
     4561 cattctctcc tctacgagta cttcaccgtc tacaacgagc tcaccaaggt caagtacgtc
     4621 acagagggta tgcgaaagcc agctttcctt tcaggcgagc agaagaaggc tatcgttgat
     4681 ctcctcttca agaccaaccg aaaggttacc gtcaagcagc tcaaggagga ctacttcaag
     4741 aagatcgagt gcttcgacag cgttgagatc agcggagttg aggatcgatt caacgcttct
     4801 ctcggtactt accacgatct cctcaagatc atcaaggaca aggacttcct ggacaacgag
     4861 gagaacgagg atatccttga ggacatcgtc ctcactctca ctcttttcga ggatcgagag
     4921 atgatcgagg agcgacttaa gacctacgct catctcttcg acgacaaggt catgaagcag
     4981 cttaagcgac gacgatatac aggttggggt cgactttctc gcaagctcat caacggtatc
     5041 cgagataagc agagcggtaa gactatcctc gacttcctca agagcgacgg ttttgctaac
     5101 cgcaacttca tgcagctcat ccacgacgat agcctcactt tcaaggagga catccagaag
     5161 gctcaggttt caggtcaagg cgattctctt cacgagcata tcgctaacct tgccggttct
     5221 cctgctatca agaagggtat cctccagaca gtcaaggtcg ttgacgagct cgttaaggtt
     5281 atgggtcgac ataagcccga gaacatcgtt atcgagatgg ctcgagagaa ccagactact
     5341 cagaagggtc agaagaactc tcgagagcga atgaagcgaa tcgaggaggg tatcaaggag
     5401 cttggttctc agatcctcaa ggagcatcct gttgagaaca ctcagctcca gaacgagaag
     5461 ctctaccttt actacctcca gaacggtcga gacatgtacg ttgatcagga gctcgacatc
     5521 aaccgactta gcgattacga cgtcgatcac atcgttcctc agtctttcct caaggacgac
     5581 tctatcgaca acaaggtcct cactcgaagc gacaagaacc gaggtaagag cgataacgtc
     5641 ccttcagagg aggtcgtcaa gaagatgaag aactattggc gccagctcct taacgctaag
     5701 ctcatcaccc agcgcaagtt cgacaacctt actaaagccg agcgaggagg tctttcagag
     5761 cttgataagg ccggtttcat caagcgacag cttgtcgaga ctcgacagat cactaagcac
     5821 gttgctcaga tcctcgactc tcgaatgaac accaagtacg acgagaacga caagctcatc
     5881 cgagaggtta aggtcatcac cctcaagtct aagctcgtta gcgacttccg caaggatttc
     5941 cagttctaca aggtccgcga gatcaacaac taccatcacg ctcacgacgc ttaccttaac
     6001 gctgttgttg gtaccgctct catcaagaag taccctaagc tcgagagcga gttcgtttac
     6061 ggcgattaca aggtctacga cgttcgaaag atgatcgcca agtcagagca ggagattggt
     6121 aaggctaccg ccaagtactt cttctacagc aacatcatga acttcttcaa gaccgagatc
     6181 acccttgcta acggagagat tcgaaagcga cctcttatcg agactaacgg agagacagga
     6241 gagatcgttt gggataaggg tcgagacttc gctacagttc gaaaggtcct ctctatgcct
     6301 caggtcaaca tcgtcaagaa gaccgaggtt cagacaggcg gtttctctaa ggagtctatc
     6361 ctccctaagc gcaactcaga taagctcatc gctcgaaaga aggattggga ccctaagaag
     6421 tacggaggtt tcgattctcc taccgttgct tactctgtcc tcgttgttgc taaggtcgag
     6481 aagggtaaga gcaagaagct caagagcgtc aaggagcttc tcggtatcac catcatggag
     6541 cgatctagct tcgagaagaa ccctatcgac ttcctcgagg ctaagggtta caaggaggtc
     6601 aagaaggacc tcatcatcaa gctccccaag tactctctct tcgagcttga gaacggtcga
     6661 aagcgaatgc ttgcttcagc cggagagctt caaaagggta acgagctcgc tctcccttct
     6721 aagtacgtca acttcctcta cctcgctagc cattacgaga agctcaaggg ttctccagag
     6781 gataacgagc agaagcagct cttcgttgag cagcacaagc actaccttga cgagatcatc
     6841 gagcagatca gcgagttctc taagcgagtt atccttgccg acgctaacct cgataaggtc
     6901 cttagcgctt acaacaagca ccgcgataag cctattcgag agcaagccga gaacatcatc
     6961 cacctcttca ctcttactaa cctcggagct ccagctgctt tcaagtactt cgacaccact
     7021 atcgatcgca agcgatacac ctctaccaag gaggttcttg acgctactct catccaccaa
     7081 tctatcaccg gtctctacga gactcgaatc gatctttctc aacttggcgg agatcctatg
     7141 gctcccaagg ctgctgacaa gaagcccgcc tccaaggctc ccgctactgc ctccaaggct
     7201 cctgagaaga aggatgccgg caagaagact gccgcctctg gtgacaagaa gaagcgctcc
     7261 aagacccgca aggagactta ctcttcttac atctacaagt aacacttaac gttactgaaa
     7321 tcatcaaaca gcttgacgaa tctggatata agatcgttgg tgtcgatgtc agctccggag
     7381 ttgagacaaa tggtgttcag gatctcgata agatacgttc atttgtccaa gcagcaaaga
     7441 gtgccttcta gtgatttaat agctccatgt caacaagaat aaaacgcgtt ttcgggttta
     7501 cctcttccag atacagctca tctgcaatgc attaatgcat tgactgcaac ctagtaacgc
     7561 ctttcaggct ccggcgaaga gaagaatagc ttagcagagc tattttcatt ttcgggagac
     7621 gagatcaagc agatcaacgg tcgtcaagag acctacgaga ctgaggaatc cgctcttggc
     7681 tccacgcgac tatatatttg tctctaattg tactttgaca tgctcctctt ctttactctg
     7741 atagcttgac tatgaaaatt ccgtcaccag cccctgggtt cgcaaagata attgcatgtt
     7801 tcttccttga actctcaagc ctacaggaca cacattcatc gtaggtataa acctcgaaat
     7861 cgtttaaact gtacacacgt gatattacat gacctggaca gagttctgcg cgaaagtcat
     7921 aacgtaaatc ccgggcggta ggtgcgtccc gggcggaagg tagttttctc gtccacccca
     7981 acgcgtttat caacctcaac tttcaacaac catcatgcca ccaaaagcgc gtaaaacaaa
     8041 gcgagatttg attgagcaag agggcaggat ccaatgcgcg attcaagaca ttaaaaatgg
     8101 aaaatttcaa aaaattgcgc ccgcagcgcg tgcatacaaa attcatccca atacaagtgt
     8161 aatgctagtg gaggtcaaca catcaatgcc tattttggtt tagtcgtcca ggcggtgagc
     8221 acaaaatttg tgtcgtttga caagatggtt catttaggca actggtcaga tcagccccac
     8281 ttgtagcagt agcggcggcg ctcgaagtgt gactcttatt agcagacagg aacgaggaca
     8341 ttattatcat ctgctgcttg gtgcacgata acttggtgcg tttgtcaagc aaggtaagtg
     8401 gacgacccgg tcataccttc ttaagttcgc ccttcctccc tttatttcag attcaatctg
     8461 acttacctat tctacccaag catccaaatg aaaaagcctg aactcaccgc gacgtctgtc
     8521 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     8581 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     8641 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     8701 ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac ctattgcatc
     8761 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     8821 ctccagccgg tcgcggaggc catggatgcg attgctgcgg ccgatcttag ccagacgagc
     8881 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     8941 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     9001 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     9061 cggcacctcg tgcatgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     9121 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     9181 atcctcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     9241 aggcatccgg agcttgcagg atcgccgcgc ctccgggcgt atatgctccg cattggtctt
     9301 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     9361 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     9421 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     9481 cgccccagca ctcgtccgag ggcaaaggaa tagagtagat gccgaccgat ccccactacc
     9541 gcattaagac ctcagcgcgg ccgcaaattt aaataaaatg aagtgaagtt cctatacttt
     9601 ctagagaata ggaacttcta tagtgagtcg aataagggcg acacaaaatt tattctaaat
     9661 gcataataaa tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat
     9721 gtataatttt gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta
     9781 attctcttta acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata
     9841 gtttaattat aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct
     9901 tttttctcat ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt
     9961 aaatattctg acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg
    10021 gtttttacgt tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga
    10081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
    10141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
    10201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
    10261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
    10321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
    10381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
    10441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
    10501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
    10561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
    10621 tctcctgttc cgaccctgcc gcttacc
//
