LOCUS       sequence-320088-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a sgRNA targeting the 3rd exon of human FMR1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Li M, Shin J, Risgaard RD, Parries MJ, Wang J, Chasman D, Liu S, Roy
            S, Bhattacharyya A, Zhao X
  TITLE     Identification of FMR1-regulated molecular networks in human 
            neurodevelopment.
  JOURNAL   Genome Res. 2020 Mar;30(3):361-374. doi: 10.1101/gr.251405.119. Epub
            2020 Mar 16.
  PUBMED    32179589
REFERENCE   2  (bases 1 to 11590)
  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: 
            "10.1101/gr.251405.119"; journalName: "Genome Res"; date: "2020-03";
            volume: "30"; issue: "3"; pages: "361-374"
COMMENT     Sequence Label: 49535-sgFMR1_E3B
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        113..324
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      344..353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             353..376
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             383..403
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             428..4528
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             428..4528
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4529..4576
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4592..4648
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4649..5248
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4649..4668)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5145..5165
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5264..5852
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5317..5337)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5735..5746)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5924..6157
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6235..6356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6272..6291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6326..6345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6396..6531
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6551..6570)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6552..6570)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6580..6597)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6580..6596)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6589..6611)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6738..7193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6825..6844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7035..7056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7219..7323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7324..7392,7393..8184)
                     /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(7542..7561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8355..8943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8844..8863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9097..9114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9231..9252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9267..9272,9273..9290,9291..9297)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9305..9321
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9310..9332
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9329..9345
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9329..9345
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9364..9384
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9366..9384
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9412..9638
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9639..9819
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9866..9991
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10484..10717
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10902..10946
                     /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"
     promoter        11124..11372
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     11124..11144
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     11295..11314
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        11394..11469
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    join(11517..11590,1..44)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
       61 acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca gtgggcagag
      121 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      181 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      241 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      301 aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca tggattacaa
      361 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      421 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      481 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      541 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
      601 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
      661 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
      721 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
      781 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
      841 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
      901 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
      961 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1021 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1081 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1141 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1201 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     1261 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1321 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1381 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1441 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1501 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1561 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     1621 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     1681 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     1741 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     1801 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     1861 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     1921 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     1981 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     2041 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2101 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2161 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     2221 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     2281 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2341 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2401 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2461 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2521 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     2581 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     2641 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     2701 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     2761 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     2821 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     2881 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     2941 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     3001 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3061 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3121 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3181 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     3241 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3301 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3361 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3421 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3481 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3541 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     3601 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     3661 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     3721 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     3781 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     3841 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     3901 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     3961 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4021 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4081 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4141 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4201 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     4261 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4321 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4381 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4441 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4501 gatcgacctg tctcagctgg gaggcgacaa gcgtcctgct gctactaaga aagctggtca
     4561 agctaagaaa aagaaagcta gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc
     4621 cggcgacgtg gaggagaacc ccggccccat gaccgagtac aagcccacgg tgcgcctcgc
     4681 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4741 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4801 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4861 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4921 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4981 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
     5041 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5101 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5161 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5221 gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gacaatcaac ctctggatta
     5281 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5341 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5401 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5461 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5521 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5581 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5641 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     5701 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5761 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5821 gagtcggatc tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag
     5881 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5941 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     6001 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6061 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6121 ctcagaccct tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat
     6181 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg
     6241 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6301 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6361 gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     6421 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6481 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt
     6541 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     6601 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     6661 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     6721 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     6781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6961 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7021 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7081 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7141 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     7201 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7261 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7321 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7381 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7441 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7501 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7561 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7621 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7681 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7741 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7801 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7861 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7921 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7981 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8041 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8101 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8161 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8221 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8281 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8341 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8401 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8461 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8521 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8581 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8641 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8701 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8761 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8821 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8881 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8941 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     9001 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     9061 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     9121 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     9181 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     9241 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     9301 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     9361 cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt
     9421 atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag
     9481 gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat
     9541 taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca
     9601 gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga
     9661 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     9721 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     9781 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     9841 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     9901 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     9961 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
    10021 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
    10081 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
    10141 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
    10201 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
    10261 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
    10321 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
    10381 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
    10441 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
    10501 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
    10561 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
    10621 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
    10681 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
    10741 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
    10801 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
    10861 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
    10921 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
    10981 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
    11041 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
    11101 tcccaacccc gaggggaccc agagagggcc tatttcccat gattccttca tatttgcata
    11161 tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat
    11221 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
    11281 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
    11341 ctttatatat cttgtggaaa ggacgaaaca ccgtattata acctacagga ggtgttttag
    11401 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
    11461 agtcggtgct tttttgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt
    11521 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
    11581 acagacatac
//
