LOCUS       sequence-496524-       13877 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of SpCas9-BFP with PSMD14 IVS2 intron in 5' UTR and 
            blasticidin resistance gene. Contains Cp36 recombination site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13877)
  AUTHORS   Arana S, Du PP, Vaughan-Jackson A, Enright N, Spees K, Valbuena R, 
            Garcia CA, Nguyen T, Venida A, Seczynska M, Bintu L, Lehner PJ, 
            Prolo LM, Bassik MC
  TITLE     Reduced Cas9 transgene silencing by incorporation of intron 
            sequences.
  JOURNAL   Nat Commun. 2025 Nov 27;16(1):10656. doi: 
            10.1038/s41467-025-65669-0.
  PUBMED    41309578
REFERENCE   2  (bases 1 to 13877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-11-27"; volume: "16(1):10656. doi"; pages: " 
            10.1038/s41467-025-65669-0"
COMMENT     Sequence Label: pSA091 PSMD14_IVS2-Cas9-BFP
FEATURES             Location/Qualifiers
     source          1..13877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        500..1709
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1546..1565
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1713..2111
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2138..2186
                     /note="synthetic polyadenylation signal"
     CDS             join(2452..2520,2521..3312)
                     /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     complement(2670..2689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3325..3461
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3613..3684
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3689..3735)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3753..3801
                     /note="synthetic polyadenylation signal"
     promoter        3942..5120
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4173..5111
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5068..5088
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7504..7513
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7510..11613
                     /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             7510..11613
                     /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             11617..11643
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11623..11649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             11662..11682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11689..11709
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11749..12447
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    12499..12620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12510..12529)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12564..12583
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      12735..12781
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      12811..12947
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    12992..13063
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(13289..13877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13369..13388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
      241 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
      301 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
      361 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
      421 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
      481 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
      541 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
      601 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
      661 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
      721 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
      781 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
      841 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
      901 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
      961 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     1021 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     1081 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     1141 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     1201 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     1261 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     1321 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     1381 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     1441 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     1501 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     1561 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     1621 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     1681 tctggccgtt tttggctttt ttgttagaca acatggccaa gcctttgtct caagaagaat
     1741 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     1801 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     1861 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     1921 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     1981 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     2041 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     2101 gggagggcta agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     2161 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     2221 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     2281 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     2341 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     2401 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     2461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     2581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     2641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     2761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     2881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3001 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3061 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3301 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     3361 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     3421 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     3481 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     3541 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     3601 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     3661 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     3721 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     3781 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccctta accgcttttg
     3841 aatgtccgtc ttagttaatc cctaattgaa cgctccaaaa ggtgacttcc aatagggatt
     3901 tattcctttt aaaattaacg gcataatcgt agatctcgtg aggctccggt gcccgtcagt
     3961 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     4021 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     4081 gcctttttcc cgagggtggg ggagaaccgt atataagtgc actagtcgcc gtgaacgttc
     4141 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     4201 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     4261 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg tggccttgcg
     4321 cttaaggagc cccttcgcct cgtgcttgag ttgtggcctg gcctgggcgc tggggccgcc
     4381 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     4441 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     4501 cgggccaaga tcagcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     4561 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     4621 gacgggggta gtctcaagct gcccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     4681 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     4741 ggccgcttcc cggccctgct gcagggagca caaaatggag gacgcggcgc tcgggagagc
     4801 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     4861 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcc agcttttgga
     4921 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     4981 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     5041 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     5101 ttccatttca ggtgtcgtga aaactacccc taaaagccaa aagatctttg tcgatcctac
     5161 catccactcg acacacccgc cagcggccgc tgccaagctt gctaggaatt ccgcgtctgc
     5221 aggatatcaa gcttgcggta cgtaaggttt aaaaatgtat tttactttta attattattt
     5281 tacggatgga tagagagata gtatgaaaat gtattatttt taacagatgt caaattatga
     5341 atgtgattga taaccagatt ttgaaaagtg gtgtgattta gaatatgtat caatatactt
     5401 cattaccaaa tgttagtatt tttattaaac ctggttatct cagcttaaag ttttaatttg
     5461 ttaaatatgt ttcctctttg ctgtgatatc taattttgct acagaaacaa atttgacctg
     5521 ccagattttc tttctctctg cttcttgcat cactcatata tattcacagc atctggagtc
     5581 tgtgtaaagt catcagaaac aacacagagg gatagataaa ctgaacacag ctgggaatca
     5641 gttttggcct agaatatata tattctaatg tatactgtat gttgtttgtt ttttctgttg
     5701 gttctttaag tttgtaaggt tgaaatggtc caaaagaaat ttttttcttc tttggtaata
     5761 tattctagag aattgcttta atattataaa atatacagga ttccccccgc cccgttttta
     5821 gcctttaaaa agtgctgtgt gttgaatgcc atatttgatt tgaggaattt caatgagtat
     5881 ttgatggaga ttttatttgt aaatgttgct attaggagga tgagttaaga tcgtgaagtg
     5941 gaaattttgt ctgtattacc acgttttctt atatgccttt tcattttaaa aaaggaattt
     6001 ttaaaattgt gagtcttata aacatgttac atgaaacaag ccagatacga aaaatatatg
     6061 attccgttta tgtaaaattc aaaagcagcc aaactaatct gtgctgttga aagttgaatt
     6121 ttacctttgc agggtggaat agactggaag cggggcttgg gtggcgtttc tgaggtgcca
     6181 ataatgttct gtttcttgaa ctaggtgatg gttacatgga tgtgttcact tttgaagatt
     6241 ccatggccat atcgtttgtt ttcttttctg tatgtgcact tgagtgaagt tcttttctgt
     6301 atgtggactt gaataaagtt tgcgatatta acattgttaa attgttacca ggattcaaca
     6361 aaggtgattg gtgggggata atatataagt acattattga gttttagttt atatagcaat
     6421 ttgaactcgt aaaacaacat ggacgaagga tgaaatgaat cttagcattt aatttagaat
     6481 ataatttgta ataataccat tttcttaagc atacacttag gaagaagagg aatttcattt
     6541 aggacatagc gattttcaag tagaaaaaat tgctttgttg tcatttttaa attacttagt
     6601 tgtactgtta ttaactttta aaatttgatt acttcttttt gttatttaag gccatataag
     6661 tacacttgat atttcttaac ttcttctgtg aggttttcaa gcttgtaaga gtatcttctg
     6721 tagttactct aaatgttaaa actaaatcag attgatatag taaatccact ttgtattggt
     6781 taactctgaa gttataattg agaagcacag ttgaatttcc ttagtgaaaa aggtgtatat
     6841 atctctacaa catctgaaac aacacaaaat atctgtgtag gttctagaac actatcttaa
     6901 tatgtttaat gtttatgcta tagtatttct tttagtataa tgagttcagt ttttaaaagc
     6961 ctttgatgct aattcagaga tccataatct gctcacaatg tctttcacat gtagttttga
     7021 tggtaactga caagctgtaa gttaattagc acagaactta ctaatgagga tgttaaaatt
     7081 taatttaaaa gtgaggattt ttatttgtac actctggacc aattgaagga aattcaatca
     7141 tacatatgat aagcatttca ggcacttttt ctacattcag gtactttatt agataacaat
     7201 aagaaaatat aattaactta atgatagcaa agttttgaaa gtatgctaag attgcttgtc
     7261 tttaaaactg gggataaccc aaaaccaaag cagtaaaata aggtctgtta attagcatgg
     7321 aagacagctt cttgaattaa tatgtggtgt taattaacag gaagtctgat gttgtgttgt
     7381 aattaccacc aatatttttg acatactgag tgactgaagg tgaattatat agatagcaat
     7441 tgaataaaca tttttgtgct taggaacgtt ttttctttgt ttctgttttc tagcgcgggc
     7501 ccggccacca tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     7561 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     7621 accgaccggc acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa
     7681 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     7741 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     7801 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     7861 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     7921 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     7981 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     8041 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     8101 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     8161 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     8221 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     8281 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     8341 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     8401 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     8461 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     8521 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     8581 ggctacgccg gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag
     8641 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     8701 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     8761 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     8821 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     8881 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     8941 ttcgaggaag tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac
     9001 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     9061 ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     9121 gccttcctga gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg
     9181 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     9241 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     9301 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     9361 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     9421 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     9481 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     9541 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     9601 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     9661 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     9721 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     9781 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     9841 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     9901 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     9961 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
    10021 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg
    10081 actcggagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
    10141 aagatgaaga actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc
    10201 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
    10261 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
    10321 cggatgaaca ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc
    10381 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
    10441 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
    10501 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
    10561 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
    10621 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
    10681 cggaagcggc ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc
    10741 cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag
    10801 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac
    10861 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
    10921 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
    10981 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
    11041 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
    11101 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
    11161 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
    11221 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
    11281 tttgtggaac agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc
    11341 aagagagtga tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac
    11401 agagacaagc ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
    11461 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
    11521 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
    11581 acacggatcg acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat
    11641 gccagcctgg gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag
    11701 cggaaagtgg acggcattgg tagtgggagc aacggcagca gcggatccag cgagctgatt
    11761 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag
    11821 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
    11881 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc
    11941 agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct
    12001 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
    12061 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
    12121 ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag
    12181 acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg
    12241 ggcgggagcc atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag
    12301 aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc
    12361 aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct
    12421 agcaaactgg ggcacaagct taattagtaa ctagcttgac tgactgagat acagcgtacc
    12481 ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
    12541 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
    12601 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
    12661 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat
    12721 ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
    12781 gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg
    12841 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
    12901 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat
    12961 cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc
    13021 actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt
    13081 gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg
    13141 gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc
    13201 gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga
    13261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
    13321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
    13381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
    13441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
    13501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
    13561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
    13621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
    13681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
    13741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
    13801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
    13861 gaaaaaaagg atctcaa
//
