LOCUS       sequence-304832-       15497 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Human expression vector containing Ubiquitous Chromatin Opening 
            Element (UCOE) upstream of EF1alpha promoter, dCas9 that is fused to
            NLS, tagBFP and a KRAB domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15497)
  AUTHORS   Chen JJ, Nathaniel DL, Raghavan P, Nelson M, Tian R, Tse E, Hong JY,
            See SK, Mok SA, Hein MY, Southworth DR, Grinberg LT, Gestwicki JE, 
            Leonetti MD, Kampmann M
  TITLE     Compromised function of the ESCRT pathway promotes endolysosomal 
            escape of tau seeds and propagation of tau aggregation.
  JOURNAL   J Biol Chem. 2019 Oct 2. pii: RA119.009432. doi: 
            10.1074/jbc.RA119.009432.
  PUBMED    31578281
REFERENCE   2  (bases 1 to 15497)
  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"; journalName: "J Biol 
            Chem. 2019 Oct 2. pii: RA119.009432. doi: 10.1074/jbc.RA119.009432."
COMMENT     Sequence Label: pMH0006
FEATURES             Location/Qualifiers
     source          1..15497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    678..803
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1300..1533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1718..1762
                     /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    2047..2164
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2896..4073
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3127..4065
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4022..4042
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4123..4132
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4129..4155,4156..4158,4159..6645,6646..6648,
                     6649..8232)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4129..8232
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8236..8262
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8242..8268
                     /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             8281..8301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8308..8328
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8368..9063
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             9109..9294
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     misc_feature    9358..9946
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9411..9431)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9829..9840)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9949..9965)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9950..9966)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10056..10289
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(10314..10332)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(10315..10332)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(10622..10641)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(10855..10874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10974..10996
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(11034..11052)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        11120..11224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11225..11293,11294..12085)
                     /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(11443..11462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12256..12844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12745..12764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12998..13015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(13085..13105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13090..13419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13270..13405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          14553..14618
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             14748..14768
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    15193..15327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(15230..15249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     15284..15303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(15495..15497,1..631)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
       61 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
      121 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
      181 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
      241 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
      301 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
      361 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
      421 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
      481 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      541 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      601 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
      661 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
      721 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
      781 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
      841 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
      901 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
      961 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     1021 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     1081 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     1141 cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     1981 agacagatcc attcgattag tgaacggatc tcgacggtat cgccaaatgg cagtattcat
     2041 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     2101 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     2161 ttttcgggtt tattacaggg acagcagaga tccagtttgg atcgataagc ttgatatcga
     2221 attcgggagg tggtccctgc agttacgcca atgataaccc ccgccagaaa aatcttagta
     2281 gccttccctt tttgttttcc gtgccccaac tcggcggatt gactcggccc cttccggaaa
     2341 cacccgaatc aacttctagt caaattattg ttcacgccgc aatgacccac ccctggcccg
     2401 cgtctgtgga actgacccct ggtgtacagg agagttcgct gctgaaagtg gtcccaaagg
     2461 ggtactagtt tttaagctcc caactccccc tcccccagcg tctggaggat tccacaccct
     2521 cgcaccgcag gggcgaggaa gtgggcggag tccggttttg gcgccagccg ctgaggctgc
     2581 caagcagaaa agccaccgct gaggagactc cggtcactgt cctcgccccg cctccccctt
     2641 ccctcccctt ggggaccacc gggcgccacg ccgcgaacgg taagtgccgc ggtcgtcggc
     2701 gcctccgccc tccccctagg gccccaattc ccagcgggcg cggcgcgcgg cccctccccc
     2761 cgccgggcgc gcgcccgctg ccccgccctt cgtggccgcc cggcgtgggc ggtgccaccc
     2821 ctccccccgg cggccccgcg cgcagctccc ggctccctcc cccttcggat gtggcttgag
     2881 ctgtaggcgc ggaggggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     2941 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
     3001 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3061 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3121 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
     3181 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
     3241 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
     3301 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
     3361 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
     3421 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
     3481 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
     3541 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
     3601 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     3661 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     3721 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     3781 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     3841 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     3901 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     3961 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     4021 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtggaattct
     4081 cacgcgtctg caggatatca agcttgcggt accgcgggcc cggccaccat ggacaagaag
     4141 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     4201 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     4261 aagaacctga tcggcgccct gctgttcgac agcggagaaa cagccgaggc cacccggctg
     4321 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     4381 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     4441 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     4501 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     4561 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     4621 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     4681 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     4741 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     4801 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg
     4861 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     4921 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     4981 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     5041 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     5101 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     5161 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacatcgat
     5221 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     5281 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     5341 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     5401 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     5461 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     5521 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     5581 ggcgccagcg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     5641 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta caacgagctg
     5701 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     5761 aaaaaagcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     5821 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     5881 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     5941 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     6001 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     6061 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     6121 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     6181 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     6241 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     6301 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     6361 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     6421 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     6481 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     6541 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     6601 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag
     6661 agctttctga aggacgactc catcgataac aaagtgctga ctcggagcga caagaaccgg
     6721 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgc
     6781 cagctgctga atgccaagct gattacccag aggaagttcg acaatctgac caaggccgag
     6841 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     6901 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     6961 gagaacgaca aactgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     7021 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     7081 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     7141 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     7201 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     7261 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     7321 acaaacggcg aaacaggcga gatcgtgtgg gataagggcc gggactttgc caccgtgcgg
     7381 aaagtgctgt ctatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     7441 ttcagcaaag agtctatcct gcccaagagg aacagcgaca agctgatcgc cagaaagaag
     7501 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     7561 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     7621 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     7681 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     7741 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     7801 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     7861 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaaacac
     7921 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     7981 gctaatctgg acaaggtgct gagcgcctac aacaagcaca gagacaagcc tatcagagag
     8041 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     8101 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     8161 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     8221 ctgggaggcg acgcctatcc ctatgacgtg cccgattatg ccagcctggg cagcggctcc
     8281 cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt
     8341 agtgggagca acggcagcag cggatccagc gagctgatta aggagaacat gcacatgaag
     8401 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
     8461 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
     8521 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
     8581 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
     8641 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     8701 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
     8761 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
     8821 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
     8881 aacatcaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
     8941 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag
     9001 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt
     9061 aatggcggtg gcggagggat ggatgctaag tcactaactg cctggtcccg gacactggtg
     9121 accttcaagg atgtatttgt ggacttcacc agggaggagt ggaagctgct ggacactgct
     9181 cagcagatcg tgtacagaaa tgtgatgctg gagaactata agaacctggt ttccttgggt
     9241 tatcagctta ctaagccaga tgtgatcctc cggttggaga agggagaaga gccctagtaa
     9301 ggccgcgact ctagagtcga cctgcaggca tgcaagcttg atatcaagct tatcgataat
     9361 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     9421 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     9481 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     9541 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     9601 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     9661 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     9721 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     9781 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     9841 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     9901 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
     9961 cctcgaggga attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt
    10021 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
    10081 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
    10141 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    10201 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    10261 cttttagtca gtgtggaaaa tctctagcag catctagaat taattccgtg tattctatag
    10321 tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct
    10381 aacgacaata tgtacaagcc taattgtgta gcatctggct tactgaagca gaccctatca
    10441 tctctctcgt aaactgccgt cagagtcggt ttggttggac gaaccttctg agtttctggt
    10501 aacgccgtcc cgcacccgga aatggtcagc gaaccaatca gcagggtcat cgctagccag
    10561 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
    10621 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
    10681 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
    10741 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
    10801 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc tcagtacaat
    10861 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
    10921 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
    10981 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
    11041 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
    11101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
    11161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
    11221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
    11281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
    11341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
    11401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
    11461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
    11521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
    11581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
    11641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
    11701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
    11761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
    11821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
    11881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
    11941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
    12001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
    12061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
    12121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
    12181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
    12241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
    12301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
    12361 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
    12421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    12481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    12541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    12601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
    12661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
    12721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
    12781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
    12841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
    12901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
    12961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
    13021 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
    13081 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
    13141 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
    13201 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
    13261 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
    13321 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
    13381 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggaca caagacaggc
    13441 ttgcgagata tgtttgagaa taccacttta tcccgcgtca gggagaggca gtgcgtaaaa
    13501 agacgcggac tcatgtgaaa tactggtttt tagtgcgcca gatctctata atctcgcgca
    13561 acctattttc ccctcgaaca ctttttaagc cgtagataaa caggctggga cacttcacat
    13621 gagcgaaaaa tacatcgtca cctgggacat gttgcagatc catgcacgta aactcgcaag
    13681 ccgactgatg ccttctgaac aatggaaagg cattattgcc gtaagccgtg gcggtctgta
    13741 ccgggtgcgt tactggcgcg tgaactgggt attcgtcatg tcgataccgt ttgtatttcc
    13801 agctacgatc acgacaacca gcgcgagctt aaagtgctga aacgcgcaga aggcgatggc
    13861 gaaggcttca tcgttattga tgacctggtg gataccggtg gtactgcggt tgcgattcgt
    13921 gaaatgtatc caaaagcgca ctttgtcacc atcttcgcaa aaccggctgg tcgtccgctg
    13981 gttgatgact atgttgttga tatcccgcaa gatacctgga ttgaacagcc gtgggatatg
    14041 ggcgtcgtat tcgtcccgcc aatctccggt cgctaatctt ttcaacgcct ggcactgccg
    14101 ggcgttgttc tttttaactt caggcgggtt acaatagttt ccagtaagta ttctggaggc
    14161 tgcatccatg acacaggcaa acctgagcga aaccctgttc aaaccccgct ttaaacatcc
    14221 tgaaacctcg acgctagtcc gccgctttaa tcacggcgca caaccgcctg tgcagtcggc
    14281 ccttgatggt aaaaccatcc ctcactggta tcgcatgatt aaccgtctga tgtggatctg
    14341 gcgcggcatt gacccacgcg aaatcctcga cgtccaggca cgtattgtga tgagcgatgc
    14401 cgaacgtacc gacgatgatt tatacgatac ggtgattggc taccgtggcg gcaactggat
    14461 ttatgagtgg gccccggatc tttgtgaagg aaccttactt ctgtggtgtg acataattgg
    14521 acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta agtgtataat
    14581 gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat ggaactgatg
    14641 aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca gaagaaatgc
    14701 catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca aaaaagaaga
    14761 gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg agtcatgctg
    14821 tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa aaagctgcac
    14881 tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg cataacagtt
    14941 ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct gctattaata
    15001 actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt aataaggaat
    15061 atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt tgtagaggtt
    15121 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
    15181 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    15241 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    15301 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
    15361 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
    15421 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
    15481 acaaacttag tagttgg
//
