LOCUS       sequence-496393-       12310 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of SpCas9-BFP and blasticidin resistance gene. 
            Contains minimal A2UCOE and Cp36 recombination site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12310)
  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 12310)
  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: pSA127 A2UCOE-Cas9-BFP
FEATURES             Location/Qualifiers
     source          1..12310
                     /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        4627..5805
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4858..5796
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5753..5773
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      5937..5946
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5943..10046
                     /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             5943..10046
                     /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             10050..10076
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10056..10082
                     /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             10095..10115
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10122..10142
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10182..10880
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    10932..11053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10943..10962)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10997..11016
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      11168..11214
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      11244..11380
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    11425..11496
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(11722..12310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11802..11821)
                     /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 agatcgcatg ccgggaggtg gtccctgcag
     3961 ttacgccaat gataaccccc gccagaaaaa tcttagtagc cttccctttt tgttttccgt
     4021 gccccaactc ggcggattga ctcggcccct tccggaaaca cccgaatcaa cttctagtca
     4081 aattattgtt cacgccgcaa tgacccaccc ctggcccgcg tctgtggaac tgacccctgg
     4141 tgtacaggag agttcgctgc tgaaagtggt cccaaagggg tactagtttt taagctccca
     4201 actccccctc ccccagcgtc tggaggattc cacaccctcg caccgcaggg gcgaggaagt
     4261 gggcggagtc cggttttggc gccagccgct gaggctgcca agcagaaaag ccaccgctga
     4321 ggagactccg gtcactgtcc tcgccccgcc tcccccttcc ctccccttgg ggaccaccgg
     4381 gcgccacgcc gcgaacggta agtgccgcgg tcgtcggcgc ctccgccctc cccctagggc
     4441 cccaattccc agcgggcgcg gcgcgcggcc cctccccccg ccgggcgcgc gcccgctgcc
     4501 ccgcccttcg tggccgcccg gcgtgggcgg tgccacccct ccccccggcg gccccgcgcg
     4561 cagctcccgg ctccctcccc cttcggatgt ggcttgagct gtaggcgcgg aggggctagc
     4621 tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4681 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     4741 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4801 agtgcactag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     4861 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     4921 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt
     4981 gggtgggaga gttcgtggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgtg
     5041 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     5101 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     5161 ttctggcaag atagtcttgt aaatgcgggc caagatcagc acactggtat ttcggttttt
     5221 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     5281 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctgcccg gcctgctctg
     5341 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     5401 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagcacaaaa
     5461 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc
     5521 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     5581 ctcgattagt tctccagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     5641 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     5701 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     5761 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaaaact acccctaaaa
     5821 gccaaaagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgctgcca
     5881 agcttgctag gaattccgcg tctgcaggat atcaagcttg cggtaccgcg ggcccggcca
     5941 ccatggacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     6001 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     6061 ggcacagcat caagaagaac ctgatcggcg ccctgctgtt cgacagcgga gaaacagccg
     6121 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     6181 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     6241 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     6301 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     6361 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     6421 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     6481 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     6541 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     6601 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggca
     6661 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     6721 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     6781 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     6841 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     6901 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     6961 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     7021 ccggctacat cgatggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     7081 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     7141 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     7201 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     7261 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     7321 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     7381 aagtggtgga caagggcgcc agcgcccaga gcttcatcga gcggatgacc aacttcgata
     7441 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     7501 tgtacaacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     7561 tgagcggcga gcagaaaaaa gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     7621 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     7681 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     7741 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     7801 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     7861 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     7921 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     7981 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     8041 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     8101 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     8161 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     8221 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     8281 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     8341 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     8401 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     8461 atatcgtgcc tcagagcttt ctgaaggacg actccatcga taacaaagtg ctgactcgga
     8521 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     8581 agaactactg gcgccagctg ctgaatgcca agctgattac ccagaggaag ttcgacaatc
     8641 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     8701 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     8761 acactaagta cgacgagaac gacaaactga tccgggaagt gaaagtgatc accctgaagt
     8821 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     8881 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     8941 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     9001 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     9061 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     9121 ggcctctgat cgagacaaac ggcgaaacag gcgagatcgt gtgggataag ggccgggact
     9181 ttgccaccgt gcggaaagtg ctgtctatgc cccaagtgaa tatcgtgaaa aagaccgagg
     9241 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gacaagctga
     9301 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     9361 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     9421 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     9481 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     9541 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     9601 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     9661 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     9721 aacagcacaa acactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     9781 tgatcctggc cgacgctaat ctggacaagg tgctgagcgc ctacaacaag cacagagaca
     9841 agcctatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     9901 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     9961 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
    10021 tcgacctgtc tcagctggga ggcgacgcct atccctatga cgtgcccgat tatgccagcc
    10081 tgggcagcgg ctcccccaag aaaaaacgca aggtggaaga tcctaagaaa aagcggaaag
    10141 tggacggcat tggtagtggg agcaacggca gcagcggatc cagcgagctg attaaggaga
    10201 acatgcacat gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat
    10261 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
    10321 gcggccctct ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga
    10381 ccttcatcaa ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct
    10441 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
    10501 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat
    10561 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt
    10621 accccgctga cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga
    10681 gccatctgat cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca
    10741 agatgcctgg cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg
    10801 agacctacgt cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac
    10861 tggggcacaa gcttaattag taactagctt gactgactga gatacagcgt accttcagct
    10921 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
    10981 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
    11041 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
    11101 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
    11161 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
    11221 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
    11281 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
    11341 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
    11401 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
    11461 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
    11521 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
    11581 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
    11641 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    11701 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
    11761 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
    11821 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
    11881 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
    11941 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
    12001 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
    12061 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
    12121 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
    12181 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
    12241 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
    12301 aggatctcaa
//
