LOCUS       sequence-382132-       13436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR vector based on Aspergillus flavus U6 promoter and terminator
            for targeting the yA gene, which contains AMA1(the HindIII-PstI 
            fragment) and ptrA selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13436)
  AUTHORS   Chang PK
  TITLE     A Simple CRISPR/Cas9 System for Efficiently Targeting Genes of 
            Aspergillus Section Flavi Species, Aspergillus nidulans, Aspergillus
            fumigatus, Aspergillus terreus, and Aspergillus niger.
  JOURNAL   Microbiol Spectr. 2023 Feb 14;11(1):e0464822. doi: 
            10.1128/spectrum.04648-22. Epub 2023 Jan 18.
  PUBMED    36651760
REFERENCE   2  (bases 1 to 13436)
  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: "Microbiol 
            Spectr."; date: "2023-02-14"; volume: "11(1):e0464822. doi"; pages: 
            " 10.1128/spectrum.04648-22. Epub 2023 Jan 18"
COMMENT     Sequence Label: pAf-CRISPR-yA
FEATURES             Location/Qualifiers
     source          1..13436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1398..1417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1651..1668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1785..1806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1821..1826,1827..1844,1845..1851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1859..1875
                     /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)."
     primer_bind     1864..1886
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1883..1899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1883..1899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     intron          3020..3135
                     /gene="gpdA"
                     /label=gpdA intron
                     /note="intron from the Aspergillus nidulans 
                     glyceraldehyde-3-phosphate dehydrogenase gene (Punt et al.,
                     1990)"
     CDS             3156..3176
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3207..7310
                     /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             3207..7310
                     /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             7323..7343
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_RNA        complement(7751..7826)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     11149..11170
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     gene            complement(11289..13296)
                     /product="pyrithiamine resistance gene of Aspergillus 
                     oryzae (Kubodera et al., 2000)"
                     /label=ptrA
                     /note="useful as a dominant selectable marker in 
                     Aspergillus"
     CDS             complement(join(11533..12179,12238..12574))
                     /codon_start=1
                     /gene="ptrA of Aspergillus oryzae"
                     /product="pyrithiamine resistance protein (Kubodera et al.,
                     2000)"
                     /label=ptrA
                     /note="useful as a dominant selectable marker in 
                     Aspergillus"
                     /translation="MSPPAAIYEPTVAATGLKGKVVVSETVPVEGASQTKLLDHFGGKW
                     DEFKFAPIRESQVSRAMTRRYFEDLDKYAESDVVIVGAGSCGLSTAYVLAKARPDLKIA
                     IVEASVSPGGGAWLGGQLFSAMVMRRPAEVFLNELGVPYEEDANPNYVVVKHASLFTST
                     LMSKVLSFPNVKLFNATAVEDLITRPTENGNPQIAGVVVNWTLVTLHHDDHSCMDPNTI
                     NAPVIISTTGHDGPFGAFCAKRLVSMGSVDKLGGMRGLDMSSAEDAIVKNTREVTKGLI
                     IGGMELSEIDGFNRMGPTFGAMVLSGVKAAEEALKVFDERQRECAE"
     CDS             12972..12983
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(13314..13382,13383..13436,1..738)
                     /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 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     1741 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     1801 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     1861 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttcga
     1921 gtaccattta attctatttg tgtttgatcg agacctaata cagcccctac aacgaccatc
     1981 aaagtcgtat agctaccagt gaggaagtgg actcaaatcg acttcagcaa catctcctgg
     2041 ataaacttta agcctaaact atacagaata agatggtgga gagcttatac cgagctccca
     2101 aatctgtcca gatcatggtt gaccggtgcc tggatcttcc tatagaatca tccttattcg
     2161 ttgacctagc tgattctgga gtgacccaga gggtcatgac ttgagcctaa aatccgccgc
     2221 ctccaccatt tgtagaaaaa tgtgacgaac tcgtgagctc tgtacagtga ccggtgactc
     2281 tttctggcat gcggagagac ggacggacgc agagagaagg gctgagtaat aagcgccact
     2341 gcgccagaca gctctggcgg ctctgaggtg cagtggatga ttattaatcc gggaccggcc
     2401 gcccctccgc cccgaagtgg aaaggctggt gtgcccctcg ttgaccaaga atctattgca
     2461 tcatcggaga atatggagct tcatcgaatc accggcagta agcgaaggag aatgtgaagc
     2521 caggggtgta tagccgtcgg cgaaatagca tgccattaac ctaggtacag aagtccaatt
     2581 gcttccgatc tggtaaaaga ttcacgagat agtaccttct ccgaagtagg tagagcgagt
     2641 acccggcgcg taagctccct aattggccca tccggcatct gtagggcgtc caaatatcgt
     2701 gcctctcctg ctttgcccgg tgtatgaaac cggaaaggcc gctcaggagc tggccagcgg
     2761 cgcagaccgg gaacacaagc tggcagtcga cccatccggt gctctgcact cgacctgctg
     2821 aggtccctca gtccctggta ggcagctttg ccccgtctgt ccgcccggtg tgtcggcggg
     2881 gttgacaagg tcgttgcgtc agtccaacat ttgttgccat attttcctgc tctccccacc
     2941 agctgctctt ttcttttctc tttcttttcc catcttcagt atattcatct tcccatccaa
     3001 gaacctttat ttcccctaag taagtacttt gctacatcca tactccatcc ttcccatccc
     3061 ttattccttt gaacctttca gttcgagctt tcccacttca tcgcagcttg actaacagct
     3121 accccgcttg agcaggcggc cgcatgatca tggccccaaa gaagaagcgg aaggtcggta
     3181 tccacggagt cccagcagcc ggccgcatgg acaagaagta ctccatcggt ctcgacatcg
     3241 gcaccaactc cgtcggttgg gccgtcatca ccgacgagta caaggtcccc tccaagaagt
     3301 tcaaggtcct cggtaacacc gaccgtcact ccatcaagaa gaacctcatc ggtgccctcc
     3361 tcttcgactc cggtgagacc gccgaggcca cccgtctcaa gcgtaccgcc cgtcgtcgtt
     3421 acacccgtcg taagaaccgt atctgctacc tccaggagat cttctccaac gagatggcta
     3481 aggtcgatga ctccttcttc caccgtctcg aggagtcctt cctcgtcgag gaggataaga
     3541 agcacgagcg tcaccccatc ttcggtaaca tcgtcgacga ggtcgcctac cacgagaagt
     3601 accccaccat ctaccacctc cgtaagaagc tggtcgactc tactgataag gctgaccttc
     3661 gtctcatcta cctcgccctc gcccacatga tcaagttccg tggtcacttc ctcatcgagg
     3721 gtgacctcaa ccctgacaac tccgatgtcg acaagctctt catccagctc gttcagactt
     3781 acaaccagct cttcgaagag aaccccatca acgcttccgg tgttgacgcc aaggccatcc
     3841 tctccgctcg tctctccaag tcccgtcgtc tcgagaacct catcgctcag ctccctggtg
     3901 agaagaagaa cggtctcttc ggtaacctta tcgccctctc tctcggtctc acccccaact
     3961 tcaagtccaa cttcgacctc gccgaggatg ccaaactcca gctctccaag gacacctacg
     4021 atgacgatct cgacaacctc ctcgcccaga tcggtgacca gtacgctgac ctcttccttg
     4081 ccgctaagaa cctctccgac gccatcctcc tctccgatat cctccgtgtc aacaccgaga
     4141 tcaccaaggc tcccctctcc gcttctatga tcaagcgtta cgatgagcac caccaggacc
     4201 tcactcttct caaggccctt gtccgtcagc agctccctga gaagtacaag gagatcttct
     4261 tcgatcagtc caagaacggt tacgccggtt acatcgacgg cggtgcttcc caggaggagt
     4321 tctacaagtt catcaagccc atcctcgaga agatggacgg caccgaggag ctcctcgtca
     4381 agctcaaccg tgaggatctc ctccgtaagc agcgtacttt cgacaacggt tccatccccc
     4441 accagatcca ccttggcgaa ctccacgcta tcctccgtcg tcaggaggac ttctacccct
     4501 tcctcaagga caaccgtgag aagatcgaga agatcctcac cttccgtatc ccctactacg
     4561 ttggccccct cgcccgtggt aactcccgtt tcgcctggat gactcgtaag tccgaagaga
     4621 ccatcactcc ctggaacttc gaggaagtcg tcgataaggg tgcctctgcc cagtccttca
     4681 tcgagcgtat gactaacttc gacaagaacc tccctaacga gaaggtcctt cctaagcact
     4741 ctctccttta cgagtacttc actgtttaca acgagctcac caaggtcaag tacgtcaccg
     4801 aaggtatgcg taagcccgcc ttcctctctg gtgagcagaa gaaggctatc gtcgacctcc
     4861 tcttcaagac caaccgtaag gttaccgtca agcagctcaa ggaagactac ttcaagaaga
     4921 tcgagtgctt cgactccgtt gagatctccg gtgtcgagga tcgtttcaac gcttccctcg
     4981 gcacctacca cgatctcctc aagatcatca aggacaagga cttcctcgac aacgaggaga
     5041 acgaggacat ccttgaggac atcgtcctca cccttaccct cttcgaggat cgtgagatga
     5101 tcgaagaacg tctcaagact tacgctcacc tcttcgacga caaggtcatg aagcagctca
     5161 agcgtcgtcg ttacactggt tggggccgtc tctcccgtaa gctgatcaac ggcatccgtg
     5221 acaagcagtc tggtaagact atcctcgact tccttaagtc cgatggtttc gccaaccgta
     5281 acttcatgca gctcatccac gacgactctc tcaccttcaa ggaggacatc cagaaggctc
     5341 aggtttccgg tcagggtgac tcccttcacg agcacatcgc taacctcgcc ggttcccccg
     5401 ctatcaagaa gggtatcctc cagaccgtta aggtcgtcga tgagctcgtc aaggttatgg
     5461 gtcgtcacaa gcccgagaac atcgttatcg agatggcccg tgagaaccag actacccaga
     5521 agggtcagaa gaactctcgt gaacgtatga agcgtatcga ggagggtatc aaggaactcg
     5581 gctcccagat ccttaaggag caccccgtcg agaacaccca gcttcagaac gagaagctct
     5641 acctctacta cctccagaac ggccgtgaca tgtacgttga tcaggagctc gacatcaacc
     5701 gtctctccga ctacgacgtc gatcacatcg tcccccagtc cttcctcaag gatgactcta
     5761 tcgacaacaa ggtcctcacc cgttccgata agaaccgtgg caagtctgac aacgtcccct
     5821 ccgaagaggt tgtcaagaag atgaagaact actggcgtca gcttctcaac gccaagctca
     5881 tcacccagcg taagttcgat aacctcacta aggctgaacg tggtggcctc tctgagctcg
     5941 acaaggccgg cttcatcaag cgtcagcttg ttgagactcg tcagatcacc aagcacgtcg
     6001 cccagattct cgactctcgc atgaacacca agtacgatga gaacgacaag ctcatccgtg
     6061 aggtcaaggt tatcactctc aagtctaagc tcgtctccga cttccgtaag gacttccagt
     6121 tctacaaggt ccgtgagatc aacaactacc accacgccca cgatgcctac ctcaacgctg
     6181 tcgttggcac tgctcttatc aagaagtacc ccaagctgga gtccgagttc gtctacggtg
     6241 actacaaggt ctacgatgtt cgtaagatga tcgctaagtc cgagcaggag atcggtaagg
     6301 ccaccgctaa gtacttcttc tactccaaca tcatgaactt cttcaagacc gagatcaccc
     6361 tcgccaacgg tgagatccgt aagcgtcccc tcatcgagac caacggtgag actggtgaga
     6421 tcgtctggga caagggtcgt gacttcgcca ctgtccgtaa ggtcctctcc atgccccagg
     6481 tcaacatcgt caagaagacc gaggtccaga ccggtggctt ctccaaggaa tctatcctcc
     6541 ccaagcgtaa ctccgacaag ctcatcgctc gcaagaagga ctgggacccc aagaagtacg
     6601 gcggtttcga ctctcccact gtcgcttact ccgtcctcgt tgtcgccaag gtcgagaagg
     6661 gtaagtctaa gaagctcaag tccgtcaagg agctcctcgg catcaccatc atggagcgtt
     6721 cctccttcga gaagaacccc atcgacttcc tcgaggccaa gggttacaag gaggtcaaga
     6781 aggacctcat catcaagctg cccaagtact ccctcttcga gcttgagaac ggtcgtaagc
     6841 gtatgctcgc ttccgccggt gagctccaga agggtaacga gctcgctctc ccctccaagt
     6901 acgtcaactt cctctacctc gcctcccact acgagaagct caagggttct cccgaagaca
     6961 acgagcagaa gcagctcttc gtcgagcagc acaagcacta ccttgatgag atcatcgagc
     7021 agatctccga gttctccaag cgtgtcatcc tcgccgacgc taacctcgat aaggtcctct
     7081 ccgcttacaa caagcaccgt gacaagccca tccgtgagca ggctgagaac atcatccacc
     7141 tcttcactct caccaacctt ggtgcccctg ctgccttcaa gtacttcgac accaccatcg
     7201 accgtaagcg ttacacctct accaaggagg tcctcgacgc cactctcatc caccagtcca
     7261 ttaccggtct ctacgagact cgtatcgacc tctctcagct cggtggtgac tcccgtgctg
     7321 accccaagaa gaagcgtaag gtctgagcta ggggaattcc ggaccggatc cacttaacgt
     7381 tactgaaatc atcaaacagc ttgacgaatc tggatataag atcgttggtg tcgatgtcag
     7441 ctccggagtt gagacaaatg gtgttcagga tctcgataag atacgttcat ttgtccaagc
     7501 agcaaagagt gccttctagt gatttaatag ctccatgtca acaagaataa aacgcgtttc
     7561 gggtttacct cttccagata cagctcatct gcaatgcatt aatgcattga ctgcaaccta
     7621 gtaacggtac cacatattta aaaaaagtct cctgccctcc gtttcgaacc cgctccaata
     7681 acaaaatagc cattctgcac ctccatacat tcctacctct atatcaagct gataaatgct
     7741 caaaaaaaaa gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt
     7801 taacttgcta tttctagctc taaaacttag tgagaatcat ttggcgactt gttcttcttt
     7861 acaatgattt atataccctt caaaagataa atctgagcag tctgattggc tgatggttta
     7921 gttgagcggg aagcgtgggc ttacataagt gccaggaaca tcaaatgcga aaaccccgct
     7981 ttgccgcctc tgaatcaccc ttagacgagg ttcgtattgt ttgtctatgt gctgagtttt
     8041 ggagtagtct agaatacgat gagtaatact caatgcatgg agtccttact ttgtggtatg
     8101 tatgtatctt ggctatcaat tgagtatctt tgtggctgag gagcacgagt tgggcgtaat
     8161 ctgggtacaa gatatataac agactggttg ttaatcacat ttcaaaagaa aaagaaataa
     8221 taaaaataga aaactgacta tgattgaatt agctggggat tgagaaagtc caggatttga
     8281 tcaaagatgg agaagacaaa gagaatgaag caaaaagcaa aacccacagt tgaattctaa
     8341 agagaactgc agattttgac ctagttaaca aggcacatca tgctatcact tccaacactc
     8401 ttgctgagcg cgttctgcct tgcctttctg gttctacgaa gtatccagtc ccgcctgcaa
     8461 cgccgcagaa acgccaaaag gtggggttgt caaccgccag ctagagctcc atcgggattc
     8521 tttggcattc cgccgttcct cggtggtcgc agataactcc ctaaattccc tgagctaact
     8581 ctaagtcgac catgccgttt atggttagcg cctcccaaaa aggaatggcc gacttagagt
     8641 tacctcttga ccgacttttt ctttcctccc ccttacattt cgttaccaca acacattcct
     8701 atatccaaac tccaggtaca taactagtcg aaatctcttt taaatctagt caagaactag
     8761 atacttaacc ttcatactgc ttagagatag cctttgaagc tcttatttcg ctctttgtct
     8821 cactctcacc ttcctcctcc tcctccaatc ctttttgctc gggactagtc caagactagt
     8881 caacgattag tatgccgagc attcgcgata acgatctccg aaagtcccca gagtactgtc
     8941 actatctcga ggcagttaag gacggggagc ttacgctgcc ggatttcaag atagtaagcc
     9001 gacccgacta gctttaagcc tagttactga ctagtccaca ggacgacaat ggcgtgcctg
     9061 atatccatcc atatgaagtc tactgccgag tgaagggatg cctcaagcgt acagtgagtc
     9121 tactgctata ctagtttctg actagttatt aactagttga caggttccct ctgccaacag
     9181 aaacatattg gtcaagcact tgaaggacaa gaactcccac ggcatggagt ttacattgca
     9241 caatggacct cccactatga aggaactgat ggaggccaaa ggcaagtcct atctagatta
     9301 cttggtgact agtccctgac tagtcaagta gcatggtatg aaggcttgtt tgaaggcact
     9361 gttctcccaa ccccgactcc taccaagaag cgcaagcgag ctgcgtaagt ttctgtgagt
     9421 ctaactagtg tattagctaa tatatagcag caccaagtcc aaggaccaca atactaaggg
     9481 tgtcgagaat tcgtgagttt cttctcccat ttcaactagt ccttgactag tcactaacta
     9541 ctttgcagca acgagggtga agctggaaat gatcaggaca atggcgaggg cccgtaagta
     9601 cagccattca atgcagacta gttgctaact aatctgtgac tagttcaagt ggtccgtacg
     9661 ccgtgcatac ccctgtgact ggtaggaatt tgagcaagcc tgtcttgccg cgcgatgaga
     9721 aaggaaaggc aagttacatt cagccccgta cctaggatca gtgctaattt ataacctcta
     9781 gccactcttt atgcagatcc gccgtgaggg tagcaaggca gctaaatcag ctggtgagaa
     9841 aggaaccata ccctgcaaga cctgtcgcaa cgcaaagggc aaaggtaagc tatccaagct
     9901 agtttgggac tagattctaa ctagtctcag caccgtgtgg ttcaaagcca tattgcgagt
     9961 tttggcgctt tttctcatcg attgacgagg caaagggagc gagtatgcaa cctcgtaagt
    10021 cagactcaga caaagaccaa ctagttattg accagtcttc tagaaggctc tgttgtggat
    10081 cttgaggccc tggagagttc ctccaacaat ccggagacaa gcaagtcctc gtcggactag
    10141 tcactaacta gactctaact agttgcagac atggataatg caaaagagac aagcaatgaa
    10201 gaaagtggta agacatttct cctttgtggt tctggactag tctttgacta gtcacagtct
    10261 taaacaagga aaatgagcat gaaaatgagg aggaaaaggc tgctgagccc gaggaagtgc
    10321 agggtgatgg cagacatggt aggttaatac cttgttagtt attgctagtc actgactagt
    10381 caataactag tctctgaaca ccttgcaatc actccgtttg cgcagctgaa cagtggtgag
    10441 gataatagta gtaagttatt ctagcttcag agttatagga gactagatac taactagtat
    10501 tagttgcaac taacctggat ctcagagact ttggcctcaa tctagaatct atctagttgt
    10561 caactagact gtggtatcat tgtcttttat tttcctagtc ctggaactag cttctaacta
    10621 gtctccctaa tatgtggctg tcttgttttt tttttttgtt tccctacccg gatatctagt
    10681 ccccttctag gttctgttaa cctctcgggc tctgatttag tttaacgcaa acctgagatt
    10741 agtttctaac tagtctctag gttttctatc cacctttaat tgtaataata aatacaagca
    10801 acgtttatac gtcaaaagca tttataaact tttaccctaa agtagcttgc ttgtgtgttt
    10861 agtttataat tagtctctta ttaatttgat gtaggtaagc ccgccacaaa tatatatttt
    10921 taacaagata ccgtggaaaa acttcgtgct atcacaaaac agtatacaaa aaataagctg
    10981 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    11041 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
    11101 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    11161 tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta taagggattt
    11221 tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    11281 ttaacaaaat ggggtgacga tgagccgctc ttgcatcttt gtttgtatta tactgtcttt
    11341 cttgttacac ataattattc tagaatgccc caccgttaca tacgggacac agccatttac
    11401 atatgcatgt ggattacgag ctaacgagtt cattcaaatc tcagaactat cacataatca
    11461 tcattcccct atcgtcaaag accgtaagac aaatccggtt catgcactga acccattcgg
    11521 gtagtgagtc atttactcag cacactcgcg ctgacgctcg tcgaacacct tcaatgcctc
    11581 ctcggcagcc ttgacaccac tgagaaccat ggcaccgaag gtagggccca tgcggttaaa
    11641 gccatcaatt tcagacagct ccataccgcc gattatcaag cccttagtaa cctcgcgggt
    11701 gttcttgacg atggcatcct cggccgagct catgtcgaga ccacgcatgc cacctagctt
    11761 gtcgacgctg cccatggaca ccaagcgctt cgcacagaag gcgccgaatg gcccatcgtg
    11821 accagtggta ctgatgatga caggagcgtt gatagtgttg gggtccatgc aggagtgatc
    11881 atcgtggtga agggtgacca gcgtccagtt gacgacaaca ccagcaatct gggggttgcc
    11941 gttctcggtc ggacgggtga tcaagtcctc aacagcggta gcattgaaga gcttgacatt
    12001 ggggaaggag agaaccttcg acatgagtgt cgaggtaaac agggaggcgt gcttgacgac
    12061 aacgtagttg gggtttgcgt cctcttcgta aggaacaccc agctcgttca ggaagacttc
    12121 cgcgggacgg cgcatgacca tagcagaaaa gagttggcca cccaaccagg caccgccacc
    12181 tattgcacgt tagttccgga aagctgagtg caaggcaatc catcatggac tactgaccag
    12241 gagagacgct ggcctcgacg atagcaatct tcaggtccgg acgagccttg gccaagacgt
    12301 acgcagtgct cagaccgcag gaaccagcac caacaatgac aacgtcactt tcagcgtact
    12361 tgtccaggtc ctcaaagtaa cgtctggtca tggcacgaga gacctggctt tcgcggatag
    12421 gggcgaactt gaactcgtcc cacttgccac cgaaatggtc caacagcttg gtctgagaag
    12481 ctccctcaac ggggacggtc tcagaaacca cgaccttacc cttgaggccg gtagcggcca
    12541 cagtgggttc gtagatggca gctggaggag acatgtttca agttgcaatg actatcatct
    12601 gttagccatt ccatcaacag gaagaacgag agaaggcatg acccttttcg ctggtattat
    12661 ccagatcaag ttttagccgt ataatctcag aacgaaccca gtccatcgat gccatgtcct
    12721 tctagactag gatcctagag tctagggccc agcttaggga gggcatgtga atgcatcgat
    12781 gactgggaac gaacaccggc ccacgccaaa gacgttacct aagatacctt gatcattgtg
    12841 agagtccagc caaaagtatt ccatgacttc catcgtatgc cctctagagg gctaatcgag
    12901 gagtgtattt acattgtcgg ttggtttggg aactatagaa gatggtcagt tattccaatc
    12961 accaaaggtt tatcgaaggg aggaagactt gttcagtttc gtccgaggac ttttggaatt
    13021 caaatctgag atagagaatt gtgtgggatg agaggaaaag gaagaagatg gtggggttca
    13081 gaaggagggg ttgagttaaa tagcatgggt tgagtcaacg tgataagggc actataccgt
    13141 atagatcagc ggcacccgat tctatccgtt ccttttgctc ctctttagct ttgaccggtg
    13201 agccggacaa gaaacaagtg aaatcatcct gacatcggcg gacgatctcc tagcttttac
    13261 atttcgttac caatgggatc ccgtaatcaa ttgcccattg aaaaaggaag agtatgagta
    13321 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
    13381 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacga
//
