LOCUS       sequence-134494-       10709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in Mammalian Cells, Cas9 Nuclease to analize 
            translocations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10709)
  AUTHORS   Mateos-Gomez PA, Gong F, Nair N, Miller KM, Lazzerini-Denchi E, 
            Sfeir A
  TITLE     Mammalian polymerase theta promotes alternative NHEJ and suppresses 
            recombination.
  JOURNAL   Nature. 2015 Feb 12;518(7538):254-7. doi: 10.1038/nature14157. Epub 
            2015 Feb 2.
  PUBMED    25642960
REFERENCE   2  (bases 1 to 10709)
  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/nature14157"; journalName: "Nature"; date: "2015-02-12- 
            12"; volume: "518"; issue: "7538"; pages: "254-7"
COMMENT     Sequence Label: pX330(puro) Rosa26-H3F3B
FEATURES             Location/Qualifiers
     source          1..10709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        704..779
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        876..1161
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1163..1440
                     /label=chicken beta-actin promoter
     intron          1441..1669
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1681..1690
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1690..1755
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1762..1782
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1807..5907
                     /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             1807..5907
                     /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             5908..5955
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5983..6000)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5989..6196
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        6210..6424
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     6345..6364
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        6435..6513
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     primer_bind     6587..6608
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    6684..6717
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6778..7275
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6813..6836)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7208..7227
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7315..7914
                     /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(7315..7334)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7811..7831
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     rep_origin      8187..8642
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8274..8293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8484..8505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8659..8678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8778..8800
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8838..8856)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8924..9028
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9029..9097,9098..9889)
                     /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(9247..9266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10060..10648
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10549..10568
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg cccgtatccg gcgagccaac gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaggta cctttcccat gattccttca tatttgcata tacgatacaa
      481 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
      541 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      601 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      661 cttgtggaaa ggacgaaaca ccgactccag tctttctaga agagttttag agctagaaat
      721 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      781 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
      841 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc
      901 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
      961 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     1021 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     1081 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     1141 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     1201 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     1261 atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
     1321 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
     1381 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
     1441 ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
     1501 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
     1561 tccgggctgt aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt
     1621 attaatgttt aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt
     1681 gccaccatgg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
     1741 gacgatgacg ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca
     1801 gcagccgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
     1861 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     1921 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     1981 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     2041 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     2101 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     2161 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     2221 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     2281 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     2341 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     2401 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     2461 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     2521 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     2581 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     2641 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     2701 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     2761 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     2821 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     2881 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     2941 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     3001 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     3061 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     3121 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     3181 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     3241 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     3301 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     3361 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     3421 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     3481 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     3541 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     3601 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     3661 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     3721 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     3781 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     3841 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     3901 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     3961 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     4021 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     4081 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     4141 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     4201 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     4261 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     4321 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     4381 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     4441 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     4501 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     4561 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     4621 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     4681 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     4741 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     4801 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     4861 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     4921 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     4981 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     5041 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     5101 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     5161 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     5221 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     5281 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     5341 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     5401 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     5461 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     5521 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     5581 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     5641 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     5701 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     5761 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     5821 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     5881 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
     5941 gcaaaaaaga aaaagtaaga attcctagag ctcgctgatc agcctcgact gtgccttcta
     6001 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     6061 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     6121 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata
     6181 gcaggcatgc tggggagcgg ccgcaattcg aacgctgacg tcatcaaccc gctccaagga
     6241 atcgcgggcc cagtgtcact aggcgggaac acccagcgcg cgtgcgccct ggcaggaaga
     6301 tggctgtgag ggacagggga gtggcgccct gcaatatttg catgtcgcta tgtgttctgg
     6361 gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtt ctgtatgaga
     6421 ccacagatct agttggaacc attcaaaaca gcatagcaag ttaaaataag gctagtccgt
     6481 tatcaacttg aaaaagtggc accgagtcgg tgctttttgg cgccgacaac cactacctga
     6541 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     6601 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taagaattcc
     6661 gatcatattc aataaccctt aatataactt cgtataatgt atgctatacg aagttattag
     6721 gtctgaagag gagtttacgt ccagccaagc ttaggatctc gacctcgaaa ttctaccggg
     6781 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctggcac
     6841 ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca
     6901 accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa
     6961 gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct
     7021 cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg
     7081 cagcggccaa tagcagcttt gctccttcgc tttctgggct cagcagctgg gaagggtggg
     7141 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     7201 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     7261 tctccgggcc tttcgacctg catccatcta gatctcgagc agctgaagct taccatgacc
     7321 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     7381 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     7441 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7501 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7561 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7621 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7681 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7741 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     7801 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     7861 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     7921 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatgcatc gatgatatca
     7981 gatccccggg atgcagaaat tgatgatcta ttaaacaata aagatgtcca ctaaaatgga
     8041 agtttttcct gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg
     8101 aaggattgga gcggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     8161 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     8221 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     8281 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     8341 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     8401 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     8461 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     8521 tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     8581 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     8641 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     8701 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     8761 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     8821 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     8881 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     8941 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     9001 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     9061 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     9121 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     9181 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     9241 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     9301 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     9361 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     9421 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     9481 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     9541 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     9601 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     9661 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     9721 taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg
     9781 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     9841 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     9901 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     9961 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    10021 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    10081 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    10141 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    10201 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    10261 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
    10321 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
    10381 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
    10441 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
    10501 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
    10561 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
    10621 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
    10681 ggccttttgc tggccttttg ctcacatgt
//
