LOCUS       sequence-204303-       10246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-dp300 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  JOURNAL   Unpublished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            "Unpublished"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-S1396R-S1397R-Hygro
FEATURES             Location/Qualifiers
     source          1..10246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     329..352
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        392..771
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        772..975
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     926..946
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1017..1036
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1017..1035
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1043..1052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1049..4450,4451..4453,4454..5050,5051,5052..5054,
                     5055..5056,5057..5059,5060..5152)
                     /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(1049..1075,1076..1078,1079..3565,3566..3568,
                     3569..5152)
                     /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             5159..5179
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5186..5251
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      6179..6188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6461..6478)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6467..6691
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6762..6778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6762..6778)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6775..6797)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6786..6802
                     /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(6810..6816,6817..6834,6835..6840))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6855..6876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6993..7010)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7164..7752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7244..7263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7923..8714,8715..8783))
                     /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     8546..8565
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8784..8888)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8939..9060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8950..8969)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9004..9023
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9091..10116)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(9091..10116)
                     /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(10134..10246,1..387))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10182..10201)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 cgcccctgag cccgcccccg gacccacccc ttcccagcct ctgagcccag aaagcgaagg
       61 agcaaagctg ctattggccg ctgccccaaa ggcctacccg cttccattgc tcagcggtgc
      121 tgtccatctg cacgagacta gtgagacgtg ctacttccat ttgtcacgtc ctgcacgacg
      181 cgagctgcgg ggcggggggg aacttcctga ctaggggagg agtagaaggt ggcgcgaagg
      241 ggccaccaaa gaacggagcc ggttggcgcc taccggtgga tgtggaatgt gtgcgaggcc
      301 agaggccact tgtgtagcgc caagtgccca gcggggctgc taaagcgcat gctccagact
      361 gccttgggaa aagcgcctcc cctacccggt tgacattgat tattgactag ttattaatag
      421 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      481 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      541 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      601 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      661 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      721 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      781 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      841 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      901 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      961 tatataagca gagctggttt agtgaaccgt cagatccgct agagatccgc ggccgctaat
     1021 acgactcact atagggagag ccgccaccat ggataaaaag tattctattg gtttagccat
     1081 cggcactaat tccgttggat gggctgtcat aaccgatgaa tacaaagtac cttcaaagaa
     1141 atttaaggtg ttggggaaca cagaccgtca ttcgattaaa aagaatctta tcggtgccct
     1201 cctattcgat agtggcgaaa cggcagaggc gactcgcctg aaacgaaccg ctcggagaag
     1261 gtatacacgt cgcaagaacc gaatatgtta cttacaagaa atttttagca atgagatggc
     1321 caaagttgac gattctttct ttcaccgttt ggaagagtcc ttccttgtcg aagaggacaa
     1381 gaaacatgaa cggcacccca tctttggaaa catagtagat gaggtggcat atcatgaaaa
     1441 gtacccaacg atttatcacc tcagaaaaaa gctagttgac tcaactgata aagcggacct
     1501 gaggttaatc tacttggctc ttgcccatat gataaagttc cgtgggcact ttctcattga
     1561 gggtgatcta aatccggaca actcggatgt cgacaaactg ttcatccagt tagtacaaac
     1621 ctataatcag ttgtttgaag agaaccctat aaatgcaagt ggcgtggatg cgaaggctat
     1681 tcttagcgcc cgcctctcta aatcccgacg gctagaaaac ctgatcgcac aattacccgg
     1741 agagaagaaa aatgggttgt tcggtaacct tatagcgctc tcactaggcc tgacaccaaa
     1801 ttttaagtcg aacttcgact tagctgaaga tgccaaattg cagcttagta aggacacgta
     1861 cgatgacgat ctcgacaatc tactggcaca aattggagat cagtatgcgg acttattttt
     1921 ggctgccaaa aaccttagcg atgcaatcct cctatctgac atactgagag ttaatactga
     1981 gattaccaag gcgccgttat ccgcttcaat gatcaaaagg tacgatgaac atcaccaaga
     2041 cttgacactt ctcaaggccc tagtccgtca gcaactgcct gagaaatata aggaaatatt
     2101 ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac ggcggagcga gtcaagagga
     2161 attctacaag tttatcaaac ccatattaga gaagatggat gggacggaag agttgcttgt
     2221 aaaactcaat cgcgaagatc tactgcgaaa gcagcggact ttcgacaacg gtagcattcc
     2281 acatcaaatc cacttaggcg aattgcatgc tatacttaga aggcaggagg atttttatcc
     2341 gttcctcaaa gacaatcgtg aaaagattga gaaaatccta acctttcgca taccttacta
     2401 tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg atgacaagaa agtccgaaga
     2461 aacgattact ccatggaatt ttgaggaagt tgtcgataaa ggtgcgtcag ctcaatcgtt
     2521 catcgagagg atgaccaact ttgacaagaa tttaccgaac gaaaaagtat tgcctaagca
     2581 cagtttactt tacgagtatt tcacagtgta caatgaactc acgaaagtta agtatgtcac
     2641 tgagggcatg cgtaaacccg cctttctaag cggagaacag aagaaagcaa tagtagatct
     2701 gttattcaag accaaccgca aagtgacagt taagcaattg aaagaggact actttaagaa
     2761 aattgaatgc ttcgattctg tcgagatctc cggggtagaa gatcgattta atgcgtcact
     2821 tggtacgtat catgacctcc taaagataat taaagataag gacttcctgg ataacgaaga
     2881 gaatgaagat atcttagaag atatagtgtt gactcttacc ctctttgaag atcgggaaat
     2941 gattgaggaa agactaaaaa catacgctca cctgttcgac gataaggtta tgaaacagtt
     3001 aaagaggcgt cgctatacgg gctggggacg attgtcgcgg aaacttatca acgggataag
     3061 agacaagcaa agtggtaaaa ctattctcga ttttctaaag agcgacggct tcgccaatag
     3121 gaactttatg cagctgatcc atgatgactc tttaaccttc aaagaggata tacaaaaggc
     3181 acaggtttcc ggacaagggg actcattgca cgaacatatt gcgaatcttg ctggttcgcc
     3241 agccatcaaa aagggcatac tccagacagt caaagtagtg gatgagctag ttaaggtcat
     3301 gggacgtcac aaaccggaaa acattgtaat cgagatggca cgcgaaaatc aaacgactca
     3361 gaaggggcaa aaaaacagtc gagagcggat gaagagaata gaagagggta ttaaagaact
     3421 gggcagccag atcttaaagg agcatcctgt ggaaaatacc caattgcaga acgagaaact
     3481 ttacctctat tacctacaaa atggaaggga catgtatgtt gatcaggaac tggacataaa
     3541 ccgtttatct gattacgacg tcgatgccat tgtaccccaa tcctttttga aggacgattc
     3601 aatcgacaat aaagtgctta cacgctcgga taagaaccga gggaaaagtg acaatgttcc
     3661 aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg cagctcctaa atgcgaaact
     3721 gataacgcaa agaaagttcg ataacttaac taaagctgag aggggtggct tgtctgaact
     3781 tgacaaggcc ggatttatta aacgtcagct cgtggaaacc cgccaaatca caaagcatgt
     3841 tgcacagata ctagattccc gaatgaatac gaaatacgac gagaacgata agctgattcg
     3901 ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg gacttcagaa aggattttca
     3961 attctataaa gttagggaga taaataacta ccaccatgcg cacgacgctt atcttaatgc
     4021 cgtcgtaggg accgcactca ttaagaaata cccgaagcta gaaagtgagt ttgtgtatgg
     4081 tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa agcgaacagg agataggcaa
     4141 ggctacagcc aaatacttct tttattctaa cattatgaat ttctttaaga cggaaatcac
     4201 tctggcaaac ggagagatac gcaaacgacc tttaattgaa accaatgggg agacaggtga
     4261 aatcgtatgg gataagggcc gggacttcgc gacggtgaga aaagttttgt ccatgcccca
     4321 agtcaacata gtaaagaaaa ctgaggtgca gaccggaggg ttttcaaagg aatcgattct
     4381 tccaaaaagg aatagtgata agctcatcgc tcgtaaaaag gactgggacc cgaaaaagta
     4441 cggtggcttc gatagcccta cagttgccta ttctgtccta gtagtggcaa aagttgagaa
     4501 gggaaaatcc aagaaactga agtcagtcaa agaattattg gggataacga ttatggagcg
     4561 ctcgtctttt gaaaagaacc ccatcgactt ccttgaggcg aaaggttaca aggaagtaaa
     4621 aaaggatctc ataattaaac taccaaagta tagtctgttt gagttagaaa atggccgaaa
     4681 acggatgttg gctagcgccg gagagcttca aaaggggaac gaactcgcac taccgtctaa
     4741 atacgtgaat ttcctgtatt tagcgtccca ttacgagaag ttgaaaggtt cacctgaaga
     4801 taacgaacag aagcaacttt ttgttgagca gcacaaacat tatctcgacg aaatcataga
     4861 gcaaatttcg gaattcagta agagagtcat cctagctgat gccaatctgg acaaagtatt
     4921 aagcgcatac aacaagcaca gggataaacc catacgtgag caggcggaaa atattatcca
     4981 tttgtttact cttaccaacc tcggcgctcc agccgcattc aagtattttg acacaacgat
     5041 agatcgcaaa cgatacactt ctaccaagga ggtgctagac gcgacactga ttcaccaatc
     5101 catcacggga ttatatgaaa ctcggataga tttgtcacag cttgggggtg acggatcccc
     5161 caagaagaag aggaaagtct cgagcgacta caaagaccat gacggtgatt ataaagatca
     5221 tgacatcgat tacaaggatg acgatgacaa ggctgcagga ggcggaggta gcaaagaaaa
     5281 taagttttct gctaaaaggt tgccatctac cagacttggc acctttctag agaatcgtgt
     5341 gaatgacttt ctgaggcgac agaatcaccc tgagtcagga gaggtcactg ttagagtagt
     5401 tcatgcttct gacaaaaccg tggaagtaaa accaggcatg aaagcaaggt ttgtggacag
     5461 tggagagatg gcagaatcct ttccataccg aaccaaagcc ctctttgcct ttgaagaaat
     5521 tgatggtgtt gacctgtgct tctttggcat gcatgttcaa gagtatggct ctgactgccc
     5581 tccacccaac cagaggagag tatatataag aaggctcgat agtgttcatt tcttccgtcc
     5641 taaatgcttg aggactgcag tctatcatga aatcctaatt ggatatttag aatatgtcaa
     5701 gaaattaggt tacacaacag ggcatatttg ggcatgtcca ccaagtgagg gagatgatta
     5761 tatcttccat tgccatcctc ctgaccagaa gatacccaag cccaagcgac tgcaggaatg
     5821 gtacaaaaaa atgcttgaca aggctgtatc agagcgtatt gtccatgact acaaggatat
     5881 ttttaaacaa gctactgaag atagattaac aagtgcaaag gaattgcctt atttcgaggg
     5941 tgatttctgg cccaatgttc tggaagaaag cattaaggaa ctggaacagg aggaagaaga
     6001 gagaaaacga gaggaaaaca ccagcaatga aagcacagat gtgaccaagg gagacagcaa
     6061 aaatgctaaa aagaagaata ataagaaaac cagcaaaaat aagagcagcc tgagtagggg
     6121 caacaagaag aaacccggga tgcccaatgt atctaacgac ctctcacaga aactatatgc
     6181 caccatggag aagcataaag aggtcttctt tgtgatccgc ctcattgctg gccctgctgc
     6241 caactccctg cctcccattg ttgatcctga tcctctcatc ccctgcgatc tgatggatgg
     6301 tcgggatgcg tttctcacgc tggcaaggga caagcacctg gagttctctt cactccgaag
     6361 agcccagtgg tccaccatgt gcatgctggt ggagctgcac acgcagagcc aggaccgctt
     6421 tgtctacacc tgcaatgaat gctgaaaacc cgctgatcag cctcgactgt gccttctagt
     6481 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6541 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6601 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6661 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     6721 tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6781 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6841 gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6901 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6961 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7021 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7081 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7141 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7201 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7261 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7321 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7381 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7441 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7501 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7561 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7621 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7681 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7741 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7801 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7861 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7921 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7981 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8041 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8101 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8161 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8221 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8281 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8341 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8401 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8461 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8521 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8581 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8641 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8701 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8761 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8821 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8881 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acgggtatac agacatgata
     8941 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     9001 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     9061 ggggtgggcg aagaactctc ggcatctact ctattccttt gccctcggac gagtgctggg
     9121 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct
     9181 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca
     9241 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg
     9301 gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa gctccggatg
     9361 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa
     9421 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc
     9481 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg
     9541 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga
     9601 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc
     9661 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
     9721 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg cgaccggctg
     9781 cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg
     9841 ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt ccggaatcgg
     9901 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
     9961 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
    10021 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
    10081 ctcgacagac gtcgcggtga gttcaggctt tttcatggtg gcgggatgca ggtcgaaagg
    10141 cccggagatg aggaagagga gaacagcgcg gcagacgtgc gcttttgaag cgtgcagaat
    10201 gccgggcctc cggaggacct tcgggcgccc gccccgcccc tgagcc
//
