LOCUS       sequence-197880-        9967 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient expression of dCas9-DNMT3a fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9967)
  AUTHORS   Stephan Beck
  TITLE     Beck lab Cas9 plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9967)
  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            "Unpublished"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-DNMT3a-Hygro
FEATURES             Location/Qualifiers
     source          1..9967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    86..207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(97..116)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     151..170
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(238..1263)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(238..1263)
                     /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(1281..1780)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1329..1348)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1722..1745
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        1785..2164
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2165..2368
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2319..2339
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2410..2429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2410..2428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2436..2445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2442..5843,5844..5846,5847..6443,6444,6445..6447,
                     6448..6449,6450..6452,6453..6545)
                     /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(2442..2468,2469..2471,2472..4958,4959..4961,
                     4962..6545)
                     /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             6552..6572
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6579..6644
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(7575..7592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7581..7805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7876..7892)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7876..7892)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7889..7911)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7900..7916
                     /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(7924..7930,7931..7948,7949..7954))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7969..7990
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8107..8124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8278..8866)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8358..8377)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9037..9828,9829..9897))
                     /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     9660..9679
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9898..9967,1..35))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
       61 gacgtcgacg ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact
      121 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      181 accattataa gctgcaataa acaagttggg gtgggcgaag aactctcggc atctactcta
      241 ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga gtacttctac
      301 acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc cgacagtccc
      361 ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat catcgaaatt
      421 gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat acgcccggag
      481 ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct gctgctccat
      541 acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg aatccccgaa
      601 catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca ggacattgtt
      661 ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg cccaaagcat
      721 cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca cagtttgcca
      781 gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag tgtattgacc
      841 gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg ccgcagcgat
      901 cgcatccatg gcctccgcga ccggctgcag aacagcgggc agttcggttt caggcaggtc
      961 ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct cgctgaattc
     1021 cccaatgtca agcacttccg gaatcgggag cgcggccgat gcaaagtgcc gataaacata
     1081 acgatctttg tagaaaccat cggcgcagct atttacccgc aggacatatc cacgccctcc
     1141 tacatcgaag ctgaaagcac gagattcttc gccctccgag agctgcatca ggtcggagac
     1201 gctgtcgaac ttttcgatca gaaacttctc gacagacgtc gcggtgagtt caggcttttt
     1261 catggtggcg ggatgcaggt cgaaaggccc ggagatgagg aagaggagaa cagcgcggca
     1321 gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg aggaccttcg ggcgcccgcc
     1381 ccgcccctga gcccgcccct gagcccgccc ccggacccac cccttcccag cctctgagcc
     1441 cagaaagcga aggagcaaag ctgctattgg ccgctgcccc aaaggcctac ccgcttccat
     1501 tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac gtgctacttc catttgtcac
     1561 gtcctgcacg acgcgagctg cggggcgggg gggaacttcc tgactagggg aggagtagaa
     1621 ggtggcgcga aggggccacc aaagaacgga gccggttggc gcctaccggt ggatgtggaa
     1681 tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc ccagcggggc tgctaaagcg
     1741 catgctccag actgccttgg gaaaagcgcc tcccctaccc ggttgacatt gattattgac
     1801 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     1861 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1921 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     1981 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     2041 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     2101 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     2161 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     2221 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     2281 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     2341 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
     2401 cgcggccgct aatacgactc actataggga gagccgccac catggataaa aagtattcta
     2461 ttggtttagc catcggcact aattccgttg gatgggctgt cataaccgat gaatacaaag
     2521 taccttcaaa gaaatttaag gtgttgggga acacagaccg tcattcgatt aaaaagaatc
     2581 ttatcggtgc cctcctattc gatagtggcg aaacggcaga ggcgactcgc ctgaaacgaa
     2641 ccgctcggag aaggtataca cgtcgcaaga accgaatatg ttacttacaa gaaattttta
     2701 gcaatgagat ggccaaagtt gacgattctt tctttcaccg tttggaagag tccttccttg
     2761 tcgaagagga caagaaacat gaacggcacc ccatctttgg aaacatagta gatgaggtgg
     2821 catatcatga aaagtaccca acgatttatc acctcagaaa aaagctagtt gactcaactg
     2881 ataaagcgga cctgaggtta atctacttgg ctcttgccca tatgataaag ttccgtgggc
     2941 actttctcat tgagggtgat ctaaatccgg acaactcgga tgtcgacaaa ctgttcatcc
     3001 agttagtaca aacctataat cagttgtttg aagagaaccc tataaatgca agtggcgtgg
     3061 atgcgaaggc tattcttagc gcccgcctct ctaaatcccg acggctagaa aacctgatcg
     3121 cacaattacc cggagagaag aaaaatgggt tgttcggtaa ccttatagcg ctctcactag
     3181 gcctgacacc aaattttaag tcgaacttcg acttagctga agatgccaaa ttgcagctta
     3241 gtaaggacac gtacgatgac gatctcgaca atctactggc acaaattgga gatcagtatg
     3301 cggacttatt tttggctgcc aaaaacctta gcgatgcaat cctcctatct gacatactga
     3361 gagttaatac tgagattacc aaggcgccgt tatccgcttc aatgatcaaa aggtacgatg
     3421 aacatcacca agacttgaca cttctcaagg ccctagtccg tcagcaactg cctgagaaat
     3481 ataaggaaat attctttgat cagtcgaaaa acgggtacgc aggttatatt gacggcggag
     3541 cgagtcaaga ggaattctac aagtttatca aacccatatt agagaagatg gatgggacgg
     3601 aagagttgct tgtaaaactc aatcgcgaag atctactgcg aaagcagcgg actttcgaca
     3661 acggtagcat tccacatcaa atccacttag gcgaattgca tgctatactt agaaggcagg
     3721 aggattttta tccgttcctc aaagacaatc gtgaaaagat tgagaaaatc ctaacctttc
     3781 gcatacctta ctatgtggga cccctggccc gagggaactc tcggttcgca tggatgacaa
     3841 gaaagtccga agaaacgatt actccatgga attttgagga agttgtcgat aaaggtgcgt
     3901 cagctcaatc gttcatcgag aggatgacca actttgacaa gaatttaccg aacgaaaaag
     3961 tattgcctaa gcacagttta ctttacgagt atttcacagt gtacaatgaa ctcacgaaag
     4021 ttaagtatgt cactgagggc atgcgtaaac ccgcctttct aagcggagaa cagaagaaag
     4081 caatagtaga tctgttattc aagaccaacc gcaaagtgac agttaagcaa ttgaaagagg
     4141 actactttaa gaaaattgaa tgcttcgatt ctgtcgagat ctccggggta gaagatcgat
     4201 ttaatgcgtc acttggtacg tatcatgacc tcctaaagat aattaaagat aaggacttcc
     4261 tggataacga agagaatgaa gatatcttag aagatatagt gttgactctt accctctttg
     4321 aagatcggga aatgattgag gaaagactaa aaacatacgc tcacctgttc gacgataagg
     4381 ttatgaaaca gttaaagagg cgtcgctata cgggctgggg acgattgtcg cggaaactta
     4441 tcaacgggat aagagacaag caaagtggta aaactattct cgattttcta aagagcgacg
     4501 gcttcgccaa taggaacttt atgcagctga tccatgatga ctctttaacc ttcaaagagg
     4561 atatacaaaa ggcacaggtt tccggacaag gggactcatt gcacgaacat attgcgaatc
     4621 ttgctggttc gccagccatc aaaaagggca tactccagac agtcaaagta gtggatgagc
     4681 tagttaaggt catgggacgt cacaaaccgg aaaacattgt aatcgagatg gcacgcgaaa
     4741 atcaaacgac tcagaagggg caaaaaaaca gtcgagagcg gatgaagaga atagaagagg
     4801 gtattaaaga actgggcagc cagatcttaa aggagcatcc tgtggaaaat acccaattgc
     4861 agaacgagaa actttacctc tattacctac aaaatggaag ggacatgtat gttgatcagg
     4921 aactggacat aaaccgttta tctgattacg acgtcgatgc cattgtaccc caatcctttt
     4981 tgaaggacga ttcaatcgac aataaagtgc ttacacgctc ggataagaac cgagggaaaa
     5041 gtgacaatgt tccaagcgag gaagtcgtaa agaaaatgaa gaactattgg cggcagctcc
     5101 taaatgcgaa actgataacg caaagaaagt tcgataactt aactaaagct gagaggggtg
     5161 gcttgtctga acttgacaag gccggattta ttaaacgtca gctcgtggaa acccgccaaa
     5221 tcacaaagca tgttgcacag atactagatt cccgaatgaa tacgaaatac gacgagaacg
     5281 ataagctgat tcgggaagtc aaagtaatca ctttaaagtc aaaattggtg tcggacttca
     5341 gaaaggattt tcaattctat aaagttaggg agataaataa ctaccaccat gcgcacgacg
     5401 cttatcttaa tgccgtcgta gggaccgcac tcattaagaa atacccgaag ctagaaagtg
     5461 agtttgtgta tggtgattac aaagtttatg acgtccgtaa gatgatcgcg aaaagcgaac
     5521 aggagatagg caaggctaca gccaaatact tcttttattc taacattatg aatttcttta
     5581 agacggaaat cactctggca aacggagaga tacgcaaacg acctttaatt gaaaccaatg
     5641 gggagacagg tgaaatcgta tgggataagg gccgggactt cgcgacggtg agaaaagttt
     5701 tgtccatgcc ccaagtcaac atagtaaaga aaactgaggt gcagaccgga gggttttcaa
     5761 aggaatcgat tcttccaaaa aggaatagtg ataagctcat cgctcgtaaa aaggactggg
     5821 acccgaaaaa gtacggtggc ttcgatagcc ctacagttgc ctattctgtc ctagtagtgg
     5881 caaaagttga gaagggaaaa tccaagaaac tgaagtcagt caaagaatta ttggggataa
     5941 cgattatgga gcgctcgtct tttgaaaaga accccatcga cttccttgag gcgaaaggtt
     6001 acaaggaagt aaaaaaggat ctcataatta aactaccaaa gtatagtctg tttgagttag
     6061 aaaatggccg aaaacggatg ttggctagcg ccggagagct tcaaaagggg aacgaactcg
     6121 cactaccgtc taaatacgtg aatttcctgt atttagcgtc ccattacgag aagttgaaag
     6181 gttcacctga agataacgaa cagaagcaac tttttgttga gcagcacaaa cattatctcg
     6241 acgaaatcat agagcaaatt tcggaattca gtaagagagt catcctagct gatgccaatc
     6301 tggacaaagt attaagcgca tacaacaagc acagggataa acccatacgt gagcaggcgg
     6361 aaaatattat ccatttgttt actcttacca acctcggcgc tccagccgca ttcaagtatt
     6421 ttgacacaac gatagatcgc aaacgataca cttctaccaa ggaggtgcta gacgcgacac
     6481 tgattcacca atccatcacg ggattatatg aaactcggat agatttgtca cagcttgggg
     6541 gtgacggatc ccccaagaag aagaggaaag tctcgagcga ctacaaagac catgacggtg
     6601 attataaaga tcatgacatc gattacaagg atgacgatga caaggctgca ggaggcggag
     6661 gtagcaacca cgaccaggaa tttgaccctc caaaggttta cccacctgtc ccagctgaga
     6721 agaggaagcc catccgggtg ctgtctctct ttgatggaat cgctacaggg ctcctggtgc
     6781 tgaaggactt gggcattcag gtggaccgct acattgcctc ggaggtgtgt gaggactcca
     6841 tcacggtggg catggtgcgg caccagggga agatcatgta cgtcggggac gtccgcagcg
     6901 tcacacagaa gcatatccag gagtggggcc cattcgatct ggtgattggg ggcagtccct
     6961 gcaatgacct ctccatcgtc aaccctgctc gcaagggcct ctacgagggc actggccggc
     7021 tcttctttga gttctaccgc ctcctgcatg atgcgcggcc caaggaggga gatgatcgcc
     7081 ccttcttctg gctctttgag aatgtggtgg ccatgggcgt tagtgacaag agggacatct
     7141 cgcgatttct cgagtccaac cctgtgatga ttgatgccaa agaagtgtca gctgcacaca
     7201 gggcccgcta cttctggggt aaccttcccg gtatgaacag gccgttggca tccactgtga
     7261 atgataagct ggagctgcag gagtgtctgg agcatggcag gatagccaag ttcagcaaag
     7321 tgaggaccat tactacgagg tcaaactcca taaagcaggg caaagaccag cattttcctg
     7381 tcttcatgaa tgagaaagag gacatcttat ggtgcactga aatggaaagg gtatttggtt
     7441 tcccagtcca ctatactgac gtctccaaca tgagccgctt ggcgaggcag agactgctgg
     7501 gccggtcatg gagcgtgcca gtcatccgcc acctcttcgc tccgctgaag gagtattgaa
     7561 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     7621 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     7681 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     7741 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     7801 tatggcttct gaggcggaaa gaaccagctg gggctcgata ccgtcgacct ctagctagag
     7861 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7921 acacaacata cgagccggaa gcataaagtg taaagcctag ggtgcctaat gagtgagcta
     7981 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8041 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8101 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8161 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8221 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8281 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8341 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8401 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8461 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8521 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8581 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8641 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8701 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8761 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     9001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     9061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     9121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     9181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9961 atttgaa
//
