LOCUS       sequence-341511-       15266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  UCOE-EF1A-Cas9-BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15266)
  AUTHORS   Hein MY, Weissman JS
  TITLE     Functional single-cell genomics of human cytomegalovirus infection.
  JOURNAL   Nat Biotechnol. 2022 Mar;40(3):391-401. doi: 
            10.1038/s41587-021-01059-3. Epub 2021 Oct 25.
  PUBMED    34697476
REFERENCE   2  (bases 1 to 15266)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41587-021-01059-3"; journalName: "Nat Biotechnol"; date: 
            "2022-03"; volume: "40"; issue: "3"; pages: "391-401"
COMMENT     Sequence Label: pMH0007
FEATURES             Location/Qualifiers
     source          1..15266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1258..1323
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1453..1473
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1898..2032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1935..1954)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1989..2008
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2200..2833
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2880..3005
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3502..3735
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3920..3964
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    4249..4366
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5098..6275
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5329..6267
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6224..6244
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      6325..6334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6331..10434
                     /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             6331..10434
                     /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             10438..10464
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10444..10470
                     /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             10483..10503
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10510..10530
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10570..11268
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    11329..11917
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11382..11402)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11800..11811)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11920..11936)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11921..11937)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12027..12260
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(12285..12303)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(12286..12303)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(12593..12612)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(12826..12845)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12945..12967
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(13005..13023)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13091..13195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13196..13264,13265..14056)
                     /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(13414..13433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14227..14815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14716..14735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14969..14986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(15056..15076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(15061..15266,1..124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(15241..15266,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
       61 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      121 caaaaagctt ggacacaaga caggcttgcg agatatgttt gagaatacca ctttatcccg
      181 cgtcagggag aggcagtgcg taaaaagacg cggactcatg tgaaatactg gtttttagtg
      241 cgccagatct ctataatctc gcgcaaccta ttttcccctc gaacactttt taagccgtag
      301 ataaacaggc tgggacactt cacatgagcg aaaaatacat cgtcacctgg gacatgttgc
      361 agatccatgc acgtaaactc gcaagccgac tgatgccttc tgaacaatgg aaaggcatta
      421 ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac tgggtattcg
      481 tcatgtcgat accgtttgta tttccagcta cgatcacgac aaccagcgcg agcttaaagt
      541 gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt attgatgacc tggtggatac
      601 cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg tcaccatctt
      661 cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt gttgatatcc cgcaagatac
      721 ctggattgaa cagccgtggg atatgggcgt cgtattcgtc ccgccaatct ccggtcgcta
      781 atcttttcaa cgcctggcac tgccgggcgt tgttcttttt aacttcaggc gggttacaat
      841 agtttccagt aagtattctg gaggctgcat ccatgacaca ggcaaacctg agcgaaaccc
      901 tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct agtccgccgc tttaatcacg
      961 gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac catccctcac tggtatcgca
     1021 tgattaaccg tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc ctcgacgtcc
     1081 aggcacgtat tgtgatgagc gatgccgaac gtaccgacga tgatttatac gatacggtga
     1141 ttggctaccg tggcggcaac tggatttatg agtgggcccc ggatctttgt gaaggaacct
     1201 tacttctgtg gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta
     1261 aatataaaat ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt
     1321 tagattccaa cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa
     1381 aacctgtttt gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa
     1441 cattctactc ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa
     1501 ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt
     1561 tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta
     1621 acctttataa gtaggcataa cagttataat cataacatac tgttttttct tactccacac
     1681 aggcatagag tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta
     1741 atttgtaaag gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat
     1801 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1861 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat
     2041 aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa
     2101 ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag
     2161 aaattgtatt tgttaaatat gtactacaaa cttagtagtt ggaagggcta attcactccc
     2221 aaagaagaca agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt
     2281 agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca
     2341 agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct
     2401 tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga
     2461 ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca
     2521 agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc
     2581 gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt
     2641 tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     2701 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     2761 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     2821 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
     2881 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     2941 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     3001 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     3061 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     3121 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     3181 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     3241 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     3301 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt
     3361 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     3421 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     3481 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     3541 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     3601 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     3661 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     3721 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     3781 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     3841 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     3901 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     3961 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     4021 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     4081 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     4141 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     4201 agtgaacgga tctcgacggt atcgccaaat ggcagtattc atccacaatt ttaaaagaaa
     4261 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     4321 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     4381 ggacagcaga gatccagttt ggatcgataa gcttgatatc gaattcggga ggtggtccct
     4441 gcagttacgc caatgataac ccccgccaga aaaatcttag tagccttccc tttttgtttt
     4501 ccgtgcccca actcggcgga ttgactcggc cccttccgga aacacccgaa tcaacttcta
     4561 gtcaaattat tgttcacgcc gcaatgaccc acccctggcc cgcgtctgtg gaactgaccc
     4621 ctggtgtaca ggagagttcg ctgctgaaag tggtcccaaa ggggtactag tttttaagct
     4681 cccaactccc cctcccccag cgtctggagg attccacacc ctcgcaccgc aggggcgagg
     4741 aagtgggcgg agtccggttt tggcgccagc cgctgaggct gccaagcaga aaagccaccg
     4801 ctgaggagac tccggtcact gtcctcgccc cgcctccccc ttccctcccc ttggggacca
     4861 ccgggcgcca cgccgcgaac ggtaagtgcc gcggtcgtcg gcgcctccgc cctcccccta
     4921 gggccccaat tcccagcggg cgcggcgcgc ggcccctccc cccgccgggc gcgcgcccgc
     4981 tgccccgccc ttcgtggccg cccggcgtgg gcggtgccac ccctcccccc ggcggccccg
     5041 cgcgcagctc ccggctccct cccccttcgg atgtggcttg agctgtaggc gcggaggggc
     5101 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     5161 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     5221 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     5281 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     5341 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     5401 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     5461 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     5521 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     5581 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     5641 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     5701 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     5761 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     5821 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     5881 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     5941 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     6001 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     6061 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     6121 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     6181 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     6241 gttcaaagtt tttttcttcc atttcaggtg tcgtggaatt ctcacgcgtc tgcaggatat
     6301 caagcttgcg gtaccgcggg cccggccacc atggacaaga agtacagcat cggcctggac
     6361 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     6421 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggcgcc
     6481 ctgctgttcg acagcggaga aacagccgag gccacccggc tgaagagaac cgccagaaga
     6541 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     6601 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     6661 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     6721 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     6781 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     6841 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     6901 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     6961 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     7021 ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg cctgaccccc
     7081 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     7141 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     7201 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     7261 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     7321 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     7381 ttcttcgacc agagcaagaa cggctacgcc ggctacatcg atggcggagc cagccaggaa
     7441 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     7501 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     7561 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     7621 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     7681 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     7741 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgccag cgcccagagc
     7801 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     7861 cacagcctgc tgtacgagta cttcaccgtg tacaacgagc tgaccaaagt gaaatacgtg
     7921 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaagc catcgtggac
     7981 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     8041 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     8101 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     8161 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     8221 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     8281 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     8341 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     8401 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     8461 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     8521 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     8581 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     8641 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     8701 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     8761 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     8821 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     8881 tccatcgata acaaagtgct gactcggagc gacaagaacc ggggcaagag cgacaacgtg
     8941 ccctccgaag aggtcgtgaa gaagatgaag aactactggc gccagctgct gaatgccaag
     9001 ctgattaccc agaggaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     9061 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     9121 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaacga caaactgatc
     9181 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     9241 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     9301 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     9361 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     9421 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     9481 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaacaggc
     9541 gagatcgtgt gggataaggg ccgggacttt gccaccgtgc ggaaagtgct gtctatgccc
     9601 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     9661 ctgcccaaga ggaacagcga caagctgatc gccagaaaga aggactggga ccctaagaag
     9721 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     9781 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     9841 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     9901 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     9961 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
    10021 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
    10081 gataatgagc agaaacagct gtttgtggaa cagcacaaac actacctgga cgagatcatc
    10141 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaggtg
    10201 ctgagcgcct acaacaagca cagagacaag cctatcagag agcaggccga gaatatcatc
    10261 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
    10321 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
    10381 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacgcctat
    10441 ccctatgacg tgcccgatta tgccagcctg ggcagcggct cccccaagaa aaaacgcaag
    10501 gtggaagatc ctaagaaaaa gcggaaagtg gacggcattg gtagtgggag caacggcagc
    10561 agcggatcca gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
    10621 gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
    10681 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
    10741 gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg catccccgac
    10801 ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagac
    10861 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
    10921 gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa gaaaacactc
    10981 ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga aggcagaaac
    11041 gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa gaccacatat
    11101 agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt ggactacaga
    11161 ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga ggtggcagtg
    11221 gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattagta aggccgcgac
    11281 tctagagtcg acctgcaggc atgcaagctt gatatcaagc ttatcgataa tcaacctctg
    11341 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
    11401 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
    11461 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
    11521 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
    11581 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
    11641 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
    11701 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
    11761 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
    11821 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
    11881 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgaggg
    11941 aattaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
    12001 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
    12061 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
    12121 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
    12181 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
    12241 agtgtggaaa atctctagca gcatctagaa ttaattccgt gtattctata gtgtcaccta
    12301 aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat
    12361 atgtacaagc ctaattgtgt agcatctggc ttactgaagc agaccctatc atctctctcg
    12421 taaactgccg tcagagtcgg tttggttgga cgaaccttct gagtttctgg taacgccgtc
    12481 ccgcacccgg aaatggtcag cgaaccaatc agcagggtca tcgctagcca gatcctctac
    12541 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
    12601 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
    12661 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
    12721 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
    12781 atgcaggagt cgcataaggg agagcgtcga atggtgcact ctcagtacaa tctgctctga
    12841 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
    12901 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
    12961 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
    13021 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
    13081 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
    13141 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
    13201 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
    13261 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
    13321 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
    13381 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
    13441 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    13501 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    13561 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    13621 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    13681 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    13741 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    13801 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    13861 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    13921 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    13981 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    14041 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    14101 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    14161 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    14221 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    14281 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    14341 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    14401 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    14461 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    14521 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    14581 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    14641 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    14701 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    14761 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    14821 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    14881 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    14941 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    15001 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctgtggaat
    15061 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    15121 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
    15181 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    15241 atcccgcccc taactccgcc cagttc
//
