LOCUS       sequence-492907-        8507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 and a sgRNA targeting the human TAF1-A loci for 
            knock-in..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8507)
  AUTHORS   Ide S, Imai R, Ochi H, Maeshima K
  TITLE     Transcriptional suppression of ribosomal DNA with phase separation.
  JOURNAL   Sci Adv. 2020 Oct 14;6(42). pii: 6/42/eabb5953. doi: 
            10.1126/sciadv.abb5953. Print 2020 Oct.
  PUBMED    33055158
REFERENCE   2  (bases 1 to 8507)
  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: "Sci Adv."; 
            date: "2020-10-14"; volume: "6(42). pii: 6/42/eabb5953. doi"; pages:
            " 10.1126/sciadv.abb5953. Print 2020 Oct"
COMMENT     Sequence Label: pX330-TAF1-A
FEATURES             Location/Qualifiers
     source          1..8507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1130..1177
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(1205..1222)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1211..1418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1427..1567
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1427..1556
                     /label=AAV2 ITR
     rep_origin      1642..2097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1729..1748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1939..1960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2114..2133)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2233..2255
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2293..2311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2379..2483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2484..2552,2553..3344)
                     /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(2702..2721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3515..4103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4004..4023
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4165..4413
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4165..4185
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4336..4355
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4435..4510
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        4607..4892
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        4894..5170
                     /label=chicken beta-actin promoter
     intron          5171..5398
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5410..5419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5419..5484
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5491..5511
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5536..8507,1..1129)
                     /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             join(5536..8507,1..1129)
                     /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"
ORIGIN
        1 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
       61 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
      121 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
      181 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
      241 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
      301 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
      361 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
      421 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
      481 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
      541 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
      601 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
      661 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
      721 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
      781 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
      841 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
      901 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
      961 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     1021 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     1081 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc
     1141 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag agctcgctga
     1201 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1261 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1321 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1381 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg
     1441 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     1501 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     1561 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     1621 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1681 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     1741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     1861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     1981 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     2041 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     2101 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     2161 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     2221 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     2281 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     2341 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2401 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2461 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2521 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2581 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2641 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2701 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2761 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2821 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2881 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2941 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3001 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3061 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3121 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3181 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     3241 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3301 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3361 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3421 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3481 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3541 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3601 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3661 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3721 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3781 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3841 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3901 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3961 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4021 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4081 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4141 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
     4201 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     4261 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     4321 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     4381 gctttatata tcttgtggaa aggacgaaac accgtagcta cacagtaagt agctgtttta
     4441 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
     4501 gagtcggtgc ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     4561 tttagcgcgt gcgccaattc tgcagacaaa tggctctaga ggtacccgtt acataactta
     4621 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaatagtaa
     4681 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     4741 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     4801 aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct acttggcagt
     4861 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     4921 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     4981 tttgtgcagc gatgggggcg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     5041 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     5101 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     5161 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     5221 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     5281 cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt
     5341 tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt
     5401 tggaccggtg ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt
     5461 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
     5521 ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
     5581 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     5641 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     5701 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     5761 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     5821 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     5881 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     5941 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     6001 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     6061 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     6121 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     6181 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     6241 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     6301 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     6361 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     6421 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     6481 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     6541 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     6601 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     6661 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     6721 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     6781 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     6841 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     6901 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     6961 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     7021 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     7081 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     7141 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     7201 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     7261 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     7321 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     7381 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     7441 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     7501 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     7561 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     7621 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     7681 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     7741 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     7801 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     7861 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     7921 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     7981 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     8041 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     8101 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     8161 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     8221 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     8281 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     8341 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     8401 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     8461 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgt
//
