LOCUS       sequence-496388-       12072 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of SpCas9-BFP with EEF2 IVS9 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 12072)
  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 12072)
  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: pSA095 EEF2_IVS9-Cas9-BFP
FEATURES             Location/Qualifiers
     source          1..12072
                     /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      5699..5708
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5705..9808
                     /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             5705..9808
                     /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             9812..9838
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9818..9844
                     /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             9857..9877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9884..9904
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9944..10642
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    10694..10815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10705..10724)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10759..10778
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      10930..10976
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      11006..11142
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    11187..11258
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(11484..12072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11564..11583)
                     /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 cgtgagcagc tgggtgatgt gcaggcccct tggcactgtt
     5281 gcgggccctg ccctgctcca agtcttgggc tcctcagtcg ggagctagaa gtaaggagca
     5341 tagaaggaat atagggtgat acttgtgcca gccacaactt ttgttcttag cccagtgagg
     5401 gcatttctgg agaagctcca agccccaggg cacagcacac ccctgctcat ctgtgtccct
     5461 gggcacaggt gggtggcggc cgtcgaccca tacccaggga ggagggcagt gattctgaag
     5521 aggatgtttc ctgacagccc cgacgcagca gtcagggcct tgtggaagcc accagtctgc
     5581 cctgggacct ggcaggaggg ccaccaactc cgggagggac ctgccctggc cctccagggt
     5641 ccagcacaac catccctgcg gcctgctgcc gccttctgct tcttccagcg cgggcccggc
     5701 caccatggac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     5761 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     5821 ccggcacagc atcaagaaga acctgatcgg cgccctgctg ttcgacagcg gagaaacagc
     5881 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     5941 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     6001 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     6061 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     6121 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     6181 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     6241 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     6301 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     6361 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     6421 caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     6481 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     6541 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     6601 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     6661 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     6721 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     6781 cgccggctac atcgatggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     6841 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     6901 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     6961 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     7021 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     7081 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     7141 ggaagtggtg gacaagggcg ccagcgccca gagcttcatc gagcggatga ccaacttcga
     7201 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     7261 cgtgtacaac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     7321 cctgagcggc gagcagaaaa aagccatcgt ggacctgctg ttcaagacca accggaaagt
     7381 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     7441 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     7501 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     7561 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     7621 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     7681 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     7741 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     7801 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     7861 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     7921 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     7981 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     8041 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     8101 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     8161 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     8221 ccatatcgtg cctcagagct ttctgaagga cgactccatc gataacaaag tgctgactcg
     8281 gagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     8341 gaagaactac tggcgccagc tgctgaatgc caagctgatt acccagagga agttcgacaa
     8401 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     8461 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     8521 gaacactaag tacgacgaga acgacaaact gatccgggaa gtgaaagtga tcaccctgaa
     8581 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     8641 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     8701 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     8761 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     8821 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     8881 gcggcctctg atcgagacaa acggcgaaac aggcgagatc gtgtgggata agggccggga
     8941 ctttgccacc gtgcggaaag tgctgtctat gccccaagtg aatatcgtga aaaagaccga
     9001 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgacaagct
     9061 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     9121 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     9181 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     9241 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     9301 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     9361 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     9421 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     9481 ggaacagcac aaacactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     9541 agtgatcctg gccgacgcta atctggacaa ggtgctgagc gcctacaaca agcacagaga
     9601 caagcctatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     9661 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     9721 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     9781 gatcgacctg tctcagctgg gaggcgacgc ctatccctat gacgtgcccg attatgccag
     9841 cctgggcagc ggctccccca agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa
     9901 agtggacggc attggtagtg ggagcaacgg cagcagcgga tccagcgagc tgattaagga
     9961 gaacatgcac atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac
    10021 atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga
    10081 gggcggccct ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa
    10141 gaccttcatc aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg
    10201 cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga
    10261 caccagcctc caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac
    10321 atccaacggc cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct
    10381 gtaccccgct gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg
    10441 gagccatctg atcgcaaaca tcaagaccac atatagatcc aagaaacccg ctaagaacct
    10501 caagatgcct ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa
    10561 cgagacctac gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa
    10621 actggggcac aagcttaatt agtaactagc ttgactgact gagatacagc gtaccttcag
    10681 ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
    10741 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
    10801 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
    10861 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta
    10921 tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc
    10981 cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct
    11041 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
    11101 ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc
    11161 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
    11221 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
    11281 atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
    11341 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
    11401 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    11461 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    11521 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    11581 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    11641 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    11701 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    11761 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    11821 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    11881 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    11941 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    12001 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    12061 aaaggatctc aa
//
