LOCUS       sequence-204503-       10246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-p300 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10246)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10246)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p-dCas9-p300-Hygro
FEATURES             Location/Qualifiers
     source          1..10246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..126
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        166..545
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        546..749
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     700..720
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     791..810
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        791..809
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      817..826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(823..4224,4225..4227,4228..4824,4825,4826..4828,
                     4829..4830,4831..4833,4834..4926)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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             join(823..849,850..852,853..3339,3340..3342,3343..4926)
                     /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             4933..4953
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4960..5025
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      5953..5962
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6235..6252)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6241..6465
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6536..6552)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6536..6552)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6549..6571)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6560..6576
                     /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)."
     promoter        complement(join(6584..6590,6591..6608,6609..6614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6629..6650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6767..6784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6938..7526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7018..7037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7697..8488,8489..8557))
                     /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     8320..8339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8558..8662)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8713..8834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8724..8743)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8778..8797
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(8865..9890)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8865..9890)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(join(9908..10246,1..161))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9956..9975)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 aggtggcgcg aaggggccac caaagaacgg agccggttgg cgcctaccgg tggatgtgga
       61 atgtgtgcga ggccagaggc cacttgtgta gcgccaagtg cccagcgggg ctgctaaagc
      121 gcatgctcca gactgccttg ggaaaagcgc ctcccctacc cggttgacat tgattattga
      181 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      241 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      301 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      361 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      421 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      481 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      541 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      601 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      661 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      721 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagagat
      781 ccgcggccgc taatacgact cactataggg agagccgcca ccatggataa aaagtattct
      841 attggtttag ccatcggcac taattccgtt ggatgggctg tcataaccga tgaatacaaa
      901 gtaccttcaa agaaatttaa ggtgttgggg aacacagacc gtcattcgat taaaaagaat
      961 cttatcggtg ccctcctatt cgatagtggc gaaacggcag aggcgactcg cctgaaacga
     1021 accgctcgga gaaggtatac acgtcgcaag aaccgaatat gttacttaca agaaattttt
     1081 agcaatgaga tggccaaagt tgacgattct ttctttcacc gtttggaaga gtccttcctt
     1141 gtcgaagagg acaagaaaca tgaacggcac cccatctttg gaaacatagt agatgaggtg
     1201 gcatatcatg aaaagtaccc aacgatttat cacctcagaa aaaagctagt tgactcaact
     1261 gataaagcgg acctgaggtt aatctacttg gctcttgccc atatgataaa gttccgtggg
     1321 cactttctca ttgagggtga tctaaatccg gacaactcgg atgtcgacaa actgttcatc
     1381 cagttagtac aaacctataa tcagttgttt gaagagaacc ctataaatgc aagtggcgtg
     1441 gatgcgaagg ctattcttag cgcccgcctc tctaaatccc gacggctaga aaacctgatc
     1501 gcacaattac ccggagagaa gaaaaatggg ttgttcggta accttatagc gctctcacta
     1561 ggcctgacac caaattttaa gtcgaacttc gacttagctg aagatgccaa attgcagctt
     1621 agtaaggaca cgtacgatga cgatctcgac aatctactgg cacaaattgg agatcagtat
     1681 gcggacttat ttttggctgc caaaaacctt agcgatgcaa tcctcctatc tgacatactg
     1741 agagttaata ctgagattac caaggcgccg ttatccgctt caatgatcaa aaggtacgat
     1801 gaacatcacc aagacttgac acttctcaag gccctagtcc gtcagcaact gcctgagaaa
     1861 tataaggaaa tattctttga tcagtcgaaa aacgggtacg caggttatat tgacggcgga
     1921 gcgagtcaag aggaattcta caagtttatc aaacccatat tagagaagat ggatgggacg
     1981 gaagagttgc ttgtaaaact caatcgcgaa gatctactgc gaaagcagcg gactttcgac
     2041 aacggtagca ttccacatca aatccactta ggcgaattgc atgctatact tagaaggcag
     2101 gaggattttt atccgttcct caaagacaat cgtgaaaaga ttgagaaaat cctaaccttt
     2161 cgcatacctt actatgtggg acccctggcc cgagggaact ctcggttcgc atggatgaca
     2221 agaaagtccg aagaaacgat tactccatgg aattttgagg aagttgtcga taaaggtgcg
     2281 tcagctcaat cgttcatcga gaggatgacc aactttgaca agaatttacc gaacgaaaaa
     2341 gtattgccta agcacagttt actttacgag tatttcacag tgtacaatga actcacgaaa
     2401 gttaagtatg tcactgaggg catgcgtaaa cccgcctttc taagcggaga acagaagaaa
     2461 gcaatagtag atctgttatt caagaccaac cgcaaagtga cagttaagca attgaaagag
     2521 gactacttta agaaaattga atgcttcgat tctgtcgaga tctccggggt agaagatcga
     2581 tttaatgcgt cacttggtac gtatcatgac ctcctaaaga taattaaaga taaggacttc
     2641 ctggataacg aagagaatga agatatctta gaagatatag tgttgactct taccctcttt
     2701 gaagatcggg aaatgattga ggaaagacta aaaacatacg ctcacctgtt cgacgataag
     2761 gttatgaaac agttaaagag gcgtcgctat acgggctggg gacgattgtc gcggaaactt
     2821 atcaacggga taagagacaa gcaaagtggt aaaactattc tcgattttct aaagagcgac
     2881 ggcttcgcca ataggaactt tatgcagctg atccatgatg actctttaac cttcaaagag
     2941 gatatacaaa aggcacaggt ttccggacaa ggggactcat tgcacgaaca tattgcgaat
     3001 cttgctggtt cgccagccat caaaaagggc atactccaga cagtcaaagt agtggatgag
     3061 ctagttaagg tcatgggacg tcacaaaccg gaaaacattg taatcgagat ggcacgcgaa
     3121 aatcaaacga ctcagaaggg gcaaaaaaac agtcgagagc ggatgaagag aatagaagag
     3181 ggtattaaag aactgggcag ccagatctta aaggagcatc ctgtggaaaa tacccaattg
     3241 cagaacgaga aactttacct ctattaccta caaaatggaa gggacatgta tgttgatcag
     3301 gaactggaca taaaccgttt atctgattac gacgtcgatg ccattgtacc ccaatccttt
     3361 ttgaaggacg attcaatcga caataaagtg cttacacgct cggataagaa ccgagggaaa
     3421 agtgacaatg ttccaagcga ggaagtcgta aagaaaatga agaactattg gcggcagctc
     3481 ctaaatgcga aactgataac gcaaagaaag ttcgataact taactaaagc tgagaggggt
     3541 ggcttgtctg aacttgacaa ggccggattt attaaacgtc agctcgtgga aacccgccaa
     3601 atcacaaagc atgttgcaca gatactagat tcccgaatga atacgaaata cgacgagaac
     3661 gataagctga ttcgggaagt caaagtaatc actttaaagt caaaattggt gtcggacttc
     3721 agaaaggatt ttcaattcta taaagttagg gagataaata actaccacca tgcgcacgac
     3781 gcttatctta atgccgtcgt agggaccgca ctcattaaga aatacccgaa gctagaaagt
     3841 gagtttgtgt atggtgatta caaagtttat gacgtccgta agatgatcgc gaaaagcgaa
     3901 caggagatag gcaaggctac agccaaatac ttcttttatt ctaacattat gaatttcttt
     3961 aagacggaaa tcactctggc aaacggagag atacgcaaac gacctttaat tgaaaccaat
     4021 ggggagacag gtgaaatcgt atgggataag ggccgggact tcgcgacggt gagaaaagtt
     4081 ttgtccatgc cccaagtcaa catagtaaag aaaactgagg tgcagaccgg agggttttca
     4141 aaggaatcga ttcttccaaa aaggaatagt gataagctca tcgctcgtaa aaaggactgg
     4201 gacccgaaaa agtacggtgg cttcgatagc cctacagttg cctattctgt cctagtagtg
     4261 gcaaaagttg agaagggaaa atccaagaaa ctgaagtcag tcaaagaatt attggggata
     4321 acgattatgg agcgctcgtc ttttgaaaag aaccccatcg acttccttga ggcgaaaggt
     4381 tacaaggaag taaaaaagga tctcataatt aaactaccaa agtatagtct gtttgagtta
     4441 gaaaatggcc gaaaacggat gttggctagc gccggagagc ttcaaaaggg gaacgaactc
     4501 gcactaccgt ctaaatacgt gaatttcctg tatttagcgt cccattacga gaagttgaaa
     4561 ggttcacctg aagataacga acagaagcaa ctttttgttg agcagcacaa acattatctc
     4621 gacgaaatca tagagcaaat ttcggaattc agtaagagag tcatcctagc tgatgccaat
     4681 ctggacaaag tattaagcgc atacaacaag cacagggata aacccatacg tgagcaggcg
     4741 gaaaatatta tccatttgtt tactcttacc aacctcggcg ctccagccgc attcaagtat
     4801 tttgacacaa cgatagatcg caaacgatac acttctacca aggaggtgct agacgcgaca
     4861 ctgattcacc aatccatcac gggattatat gaaactcgga tagatttgtc acagcttggg
     4921 ggtgacggat cccccaagaa gaagaggaaa gtctcgagcg actacaaaga ccatgacggt
     4981 gattataaag atcatgacat cgattacaag gatgacgatg acaaggctgc aggaggcgga
     5041 ggtagcaaag aaaataagtt ttctgctaaa aggttgccat ctaccagact tggcaccttt
     5101 ctagagaatc gtgtgaatga ctttctgagg cgacagaatc accctgagtc aggagaggtc
     5161 actgttagag tagttcatgc ttctgacaaa accgtggaag taaaaccagg catgaaagca
     5221 aggtttgtgg acagtggaga gatggcagaa tcctttccat accgaaccaa agccctcttt
     5281 gcctttgaag aaattgatgg tgttgacctg tgcttctttg gcatgcatgt tcaagagtat
     5341 ggctctgact gccctccacc caaccagagg agagtataca tatcttacct cgatagtgtt
     5401 catttcttcc gtcctaaatg cttgaggact gcagtctatc atgaaatcct aattggatat
     5461 ttagaatatg tcaagaaatt aggttacaca acagggcata tttgggcatg tccaccaagt
     5521 gagggagatg attatatctt ccattgccat cctcctgacc agaagatacc caagcccaag
     5581 cgactgcagg aatggtacaa aaaaatgctt gacaaggctg tatcagagcg tattgtccat
     5641 gactacaagg atatttttaa acaagctact gaagatagat taacaagtgc aaaggaattg
     5701 ccttatttcg agggtgattt ctggcccaat gttctggaag aaagcattaa ggaactggaa
     5761 caggaggaag aagagagaaa acgagaggaa aacaccagca atgaaagcac agatgtgacc
     5821 aagggagaca gcaaaaatgc taaaaagaag aataataaga aaaccagcaa aaataagagc
     5881 agcctgagta ggggcaacaa gaagaaaccc gggatgccca atgtatctaa cgacctctca
     5941 cagaaactat atgccaccat ggagaagcat aaagaggtct tctttgtgat ccgcctcatt
     6001 gctggccctg ctgccaactc cctgcctccc attgttgatc ctgatcctct catcccctgc
     6061 gatctgatgg atggtcggga tgcgtttctc acgctggcaa gggacaagca cctggagttc
     6121 tcttcactcc gaagagccca gtggtccacc atgtgcatgc tggtggagct gcacacgcag
     6181 agccaggacc gctttgtcta cacctgcaat gaatgctgaa aacccgctga tcagcctcga
     6241 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     6301 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     6361 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     6421 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     6481 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     6541 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     6601 gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc
     6661 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     6721 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     6781 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     6841 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     6901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     7021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     7081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     7141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     7201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     7261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     7321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     7381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     7441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     7501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     7561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     7621 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     7681 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     7741 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     7801 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     7861 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     7921 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7981 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8041 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8101 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     8161 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     8221 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     8281 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     8341 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     8401 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     8461 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     8521 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     8581 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     8641 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacgggt
     8701 atacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     8761 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     8821 gcaataaaca agttggggtg ggcgaagaac tctcggcatc tactctattc ctttgccctc
     8881 ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc
     8941 cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg
     9001 acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag
     9061 ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct
     9121 gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac
     9181 ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc
     9241 cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc
     9301 gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga
     9361 gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg
     9421 ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt
     9481 ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc
     9541 tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca
     9601 ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc
     9661 acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag
     9721 aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg
     9781 aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt
     9841 tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat ggtggcggga
     9901 tgcaggtcga aaggcccgga gatgaggaag aggagaacag cgcggcagac gtgcgctttt
     9961 gaagcgtgca gaatgccggg cctccggagg accttcgggc gcccgccccg cccctgagcc
    10021 cgcccctgag cccgcccccg gacccacccc ttcccagcct ctgagcccag aaagcgaagg
    10081 agcaaagctg ctattggccg ctgccccaaa ggcctacccg cttccattgc tcagcggtgc
    10141 tgtccatctg cacgagacta gtgagacgtg ctacttccat ttgtcacgtc ctgcacgacg
    10201 cgagctgcgg ggcggggggg aacttcctga ctaggggagg agtaga
//
