LOCUS       sequence-496522-       13948 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of SpCas9-BFP with EIF4A3 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 13948)
  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 13948)
  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: pSA090 EIF4A3_IVS2-Cas9-BFP
FEATURES             Location/Qualifiers
     source          1..13948
                     /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      7575..7584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7581..11684
                     /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             7581..11684
                     /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             11688..11714
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11694..11720
                     /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             11733..11753
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11760..11780
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11820..12518
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    12570..12691
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12581..12600)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12635..12654
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      12806..12852
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      12882..13018
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    13063..13134
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(13360..13948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13440..13459)
                     /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 cgtaagttgt cctggggtgc ggcgcagggc tggatgtaaa
     5281 tttgggcagc gcagtgcagt tagatgctct gggatgactg tttccctcag cttgtacacg
     5341 ctcagttcca acactttccc tgactttagc cacagggttt aatacttttt taaatttgtg
     5401 ttctctactg tggaaagctt tcattttctt tttgagatgg agtctcgctc tgttgcccag
     5461 gctggagtgc agtggtgcca tctcggctca ctgcaacctt tgcctctcag tttcaagcta
     5521 ttctcctgcc tcaccctccc aagtagctgg gattacaggc atgcaccacc atgcccgact
     5581 aattttcata tttttagtag agacgggggt ttcaccatgt tggtcaggct ggtctcaaac
     5641 tcctgacctc aggtgatcca ccctccttgg cctcctaaag tgctgggatt tcaggcatga
     5701 gttactgcgc ccggccaagg ctttcatttt cttagccagc tttgcttagt ggtttgctgg
     5761 catgtggaat tgtatataac acaatttcgg aaaaacagac tgtgcttttc ctctgctctc
     5821 acaccaccac aacagtcatc agcacagaag aagacttctg tgagcagagg tgtggggctt
     5881 ttccagctgg gtgtcctcca gttcaactca caccacctgc ctggagatag gccatatcct
     5941 gcaggttgac ggctcagtcc ccatgactac cctcttttca gacaccagtt ataagttcgg
     6001 atctctagaa cttctgacca agcagcttca agttgggttc ccaggatccc ctctttaggt
     6061 tggattaatt tgctggaaca gcagctcaaa gaactcaggg aaacacattt agtggtttat
     6121 tataaagggt actgcagagg atacagatat ggggggaggg gtgcagcctc caaggggcgg
     6181 gtggtcacct cccggaacct ccttgtgttc cactgtctgt gttcagccat ctagaagctc
     6241 ttcgaaccca gtcctcttgg gtttttatgg aaaattccat gaggtcagca ttctttcccc
     6301 cataagggaa tagggcagga gcttttctgg gataatacca taatgctttt taaaaaaaaa
     6361 tttgaaattt aaatttttta catttttcga gacagggtct tactctgtct cccagactgg
     6421 agtgcagtcg tgtgattaca gctcatagca gcctcaagcg atcctcccac ctcaccctcc
     6481 caagtagctg ggactacagg cacacatcac catgcccaat taatttttgt attttctgta
     6541 gagacagggt tttgttgtgt tgcccaggct gctctcaaac tcctggcaat ctgtctgcct
     6601 tggcttccca aagccttggg attacaggcg tgagccactg tgcccggcct aaacattttt
     6661 tttgttaaga tatggggcct cacttcgtca cccaggcggg aatgcagtgg tgcagtcata
     6721 gctcactgca acctctaact tcgaggatta aggaatcgtc ccgcctcaac ctcccaagta
     6781 gctgggactg ccagaatgtg ccaccgttcc tggttagttt tttgttttgt ttagaaatgg
     6841 ggtctcttgc tgtattgccc aggctggtct cgaattcctg gcttcaatta atgctgcctc
     6901 tgcgtcccag agtgttggga atacaggcct gagccaccac gccgtgacat gagggtcttt
     6961 atgactcaca atctgaaagg cagggaagat tagagccctg ctgtggagca ggtgaaagga
     7021 gagcagaagg tcagagagat tgtttcctga ggcctaaccc atcaatttaa taagactgta
     7081 ataagggcta tgggagctgt gagccaggaa ctatggatga aaacttgtat atagccatat
     7141 ctcctagcac cacaaatcct cttttaatag ctttattcct cgtttgccta atttcaaaaa
     7201 agactcattt acacaaaaat ttctttggaa gagagagtta gtccaaaatc ctagaaggaa
     7261 aagagttgta tttagataaa ggggagtcct cctttttctg acacctatga aacacctctt
     7321 gaagttagag agtctgtggc tccattgccc ccgtgcagaa gccatgtcta ctctctttct
     7381 tcatggacac ttaacaagct gtgacactgg gcccgagtcc tgcctcctcg cagttaagat
     7441 gttattaggt acgggggctt gctggaagaa ggtacctaag tgcctctgga gcccaggcct
     7501 tggagtgcag ctgtggcgtg gcctgtcctc cctctaatcc cactgctttt gtttccttga
     7561 atagcgcggg cccggccacc atggacaaga agtacagcat cggcctggac atcggcacca
     7621 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
     7681 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggcgcc ctgctgttcg
     7741 acagcggaga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     7801 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     7861 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     7921 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     7981 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     8041 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     8101 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     8161 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     8221 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     8281 agaatggcct gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     8341 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     8401 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     8461 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     8521 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     8581 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     8641 agagcaagaa cggctacgcc ggctacatcg atggcggagc cagccaggaa gagttctaca
     8701 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     8761 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     8821 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     8881 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     8941 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     9001 ccccctggaa cttcgaggaa gtggtggaca agggcgccag cgcccagagc ttcatcgagc
     9061 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     9121 tgtacgagta cttcaccgtg tacaacgagc tgaccaaagt gaaatacgtg accgagggaa
     9181 tgagaaagcc cgccttcctg agcggcgagc agaaaaaagc catcgtggac ctgctgttca
     9241 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     9301 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     9361 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     9421 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     9481 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     9541 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     9601 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     9661 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     9721 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     9781 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     9841 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     9901 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     9961 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
    10021 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
    10081 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgata
    10141 acaaagtgct gactcggagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
    10201 aggtcgtgaa gaagatgaag aactactggc gccagctgct gaatgccaag ctgattaccc
    10261 agaggaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
    10321 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
    10381 tcctggactc ccggatgaac actaagtacg acgagaacga caaactgatc cgggaagtga
    10441 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
    10501 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
    10561 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
    10621 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
    10681 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
    10741 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaacaggc gagatcgtgt
    10801 gggataaggg ccgggacttt gccaccgtgc ggaaagtgct gtctatgccc caagtgaata
    10861 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
    10921 ggaacagcga caagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
    10981 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
    11041 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
    11101 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
    11161 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
    11221 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
    11281 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
    11341 agaaacagct gtttgtggaa cagcacaaac actacctgga cgagatcatc gagcagatca
    11401 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaggtg ctgagcgcct
    11461 acaacaagca cagagacaag cctatcagag agcaggccga gaatatcatc cacctgttta
    11521 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
    11581 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
    11641 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacgcctat ccctatgacg
    11701 tgcccgatta tgccagcctg ggcagcggct cccccaagaa aaaacgcaag gtggaagatc
    11761 ctaagaaaaa gcggaaagtg gacggcattg gtagtgggag caacggcagc agcggatcca
    11821 gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc
    11881 atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga
    11941 gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct
    12001 tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc
    12061 agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc
    12121 tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca
    12181 gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg
    12241 ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc
    12301 tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga
    12361 aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa
    12421 tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact
    12481 gcgacctccc tagcaaactg gggcacaagc ttaattagta actagcttga ctgactgaga
    12541 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
    12601 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
    12661 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
    12721 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
    12781 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
    12841 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
    12901 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
    12961 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
    13021 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
    13081 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
    13141 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
    13201 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
    13261 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
    13321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    13381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    13441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    13501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    13561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    13621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    13681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    13741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    13801 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    13861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    13921 gattacgcgc agaaaaaaag gatctcaa
//
