LOCUS       sequence-496518-       12476 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of SpCas9-BFP with HBB IVS2 intron in 5' UTR and 
            blasticidin resistance gene. Flanked by cHS4 core insulators. 
            Contains Cp36 recombination site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12476)
  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 12476)
  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: pSA144 HBB_IVS2-Cas9-BFP with core cHS4 insula...
FEATURES             Location/Qualifiers
     source          1..12476
                     /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        4185..5363
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4416..5354
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5311..5331
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6345..6354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6351..10454
                     /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             6351..10454
                     /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             10458..10484
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10464..10490
                     /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             10503..10523
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10530..10550
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10590..11288
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     polyA_signal    11340..11461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11351..11370)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11405..11424
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(11888..12476)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11968..11987)
                     /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 cttttccccg tatcccccca
     3961 ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc
     4021 ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg
     4081 accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac
     4141 atccctgggg gctttggggg ggggctgtcc ctgctagctc gtgaggctcc ggtgcccgtc
     4201 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     4261 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     4321 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcactagtc gccgtgaacg
     4381 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     4441 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     4501 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgtggcctt
     4561 gcgcttaagg agccccttcg cctcgtgctt gagttgtggc ctggcctggg cgctggggcc
     4621 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     4681 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     4741 atgcgggcca agatcagcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     4801 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     4861 tcggacgggg gtagtctcaa gctgcccggc ctgctctggt gcctggcctc gcgccgccgt
     4921 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     4981 gatggccgct tcccggccct gctgcaggga gcacaaaatg gaggacgcgg cgctcgggag
     5041 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     5101 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tccagctttt
     5161 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     5221 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     5281 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     5341 ttcttccatt tcaggtgtcg tgaaaactac ccctaaaagc caaaagatct ttgtcgatcc
     5401 taccatccac tcgacacacc cgccagcggc cgctgccaag cttgctagga attccgcgtc
     5461 tgcaggatat caagcttgcg gtacgtgagt ctatgggacg cttgatgttt tctttcccct
     5521 tcttttctat ggttaagttc atgtcatagg aaggggataa gtaacagggt acagtttaga
     5581 atgggaaaca gacgaatgat tgcatcagtg tggaagtctc aggatcgttt tagtttcttt
     5641 tatttgctgt tcataacaat tgttttcttt tgtttaattc ttgctttctt tttttttctt
     5701 ctccgcaatt tttactatta tacttaatgc cttaacattg tgtataacaa aaggaaatat
     5761 ctctgagata cattaagtaa cttaaaaaaa aactttacac agtctgccta gtacattact
     5821 atttggaata tatgtgtgct tatttgcata ttcataatct ccctacttta ttttctttta
     5881 tttttaattg atacataatc attatacata tttatgggtt aaagtgtaat gttttaatat
     5941 gtgtacacat attgaccaaa tcagggtaat tttgcatttg taattttaaa aaatgctttc
     6001 ttcttttaat atactttttt gtttatctta tttctaatac tttccctaat ctctttcttt
     6061 cagggcaata atgatacaat gtatcatgcc tctttgcacc attctaaaga ataacagtga
     6121 taatttctgg gttaaggcaa tagcaatatc tctgcatata aatatttctg catataaatt
     6181 gtaactgatg taagaggttt catattgcta atagcagcta caatccagct accattctgc
     6241 ttttatttta tggttgggat aaggctggat tattctgagt ccaagctagg cccttttgct
     6301 aatcatgttc atacctctta tcttcctccc acagcgcggg cccggccacc atggacaaga
     6361 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
     6421 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     6481 agaagaacct gatcggcgcc ctgctgttcg acagcggaga aacagccgag gccacccggc
     6541 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     6601 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     6661 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     6721 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     6781 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     6841 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     6901 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     6961 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     7021 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc
     7081 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     7141 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     7201 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     7261 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     7321 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     7381 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacatcg
     7441 atggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     7501 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     7561 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     7621 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     7681 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     7741 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     7801 agggcgccag cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     7861 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tacaacgagc
     7921 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     7981 agaaaaaagc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     8041 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     8101 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     8161 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     8221 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     8281 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     8341 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     8401 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     8461 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     8521 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     8581 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     8641 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     8701 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     8761 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     8821 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     8881 agagctttct gaaggacgac tccatcgata acaaagtgct gactcggagc gacaagaacc
     8941 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     9001 gccagctgct gaatgccaag ctgattaccc agaggaagtt cgacaatctg accaaggccg
     9061 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     9121 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     9181 acgagaacga caaactgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     9241 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     9301 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     9361 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     9421 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     9481 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     9541 agacaaacgg cgaaacaggc gagatcgtgt gggataaggg ccgggacttt gccaccgtgc
     9601 ggaaagtgct gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     9661 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga caagctgatc gccagaaaga
     9721 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     9781 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     9841 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     9901 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     9961 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
    10021 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
    10081 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaaac
    10141 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
    10201 acgctaatct ggacaaggtg ctgagcgcct acaacaagca cagagacaag cctatcagag
    10261 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
    10321 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
    10381 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
    10441 agctgggagg cgacgcctat ccctatgacg tgcccgatta tgccagcctg ggcagcggct
    10501 cccccaagaa aaaacgcaag gtggaagatc ctaagaaaaa gcggaaagtg gacggcattg
    10561 gtagtgggag caacggcagc agcggatcca gcgagctgat taaggagaac atgcacatga
    10621 agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag
    10681 gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc
    10741 ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc
    10801 acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga
    10861 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
    10921 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg
    10981 tgatgcagaa gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg
    11041 gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg
    11101 caaacatcaa gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg
    11161 tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg
    11221 agcagcacga ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc
    11281 ttaattagta actagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
    11341 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
    11401 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
    11461 tagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
    11521 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
    11581 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
    11641 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aattaattaa
    11701 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
    11761 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
    11821 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
    11881 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
    11941 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
    12001 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
    12061 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
    12121 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
    12181 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    12241 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    12301 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
    12361 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    12421 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaa
//
