LOCUS       sequence-204468-        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  JOURNAL   Unpublished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            "Unpublished"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"
     promoter        complement(160..659)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(208..227)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     601..624
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     enhancer        664..1043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1044..1247
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1198..1218
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1289..1308
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1289..1307
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1315..1324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1321..4722,4723..4725,4726..5322,5323,5324..5326,
                     5327..5328,5329..5331,5332..5424)
                     /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(1321..1347,1348..1350,1351..3837,3838..3840,
                     3841..5424)
                     /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             5431..5451
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5458..5523
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(6454..6471)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6460..6684
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6755..6771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6755..6771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6768..6790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6779..6795
                     /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(6803..6809,6810..6827,6828..6833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6848..6869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6986..7003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7157..7745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7237..7256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7916..8707,8708..8776))
                     /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     8539..8558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8777..8881)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8932..9053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8943..8962)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8997..9016
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(9084..9967,1..142))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(join(9084..9967,1..142))
                     /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"
ORIGIN
        1 ggacatatcc acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga
       61 gctgcatcag gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg
      121 cggtgagttc aggctttttc atggtggcgg gatgcaggtc gaaaggcccg gagatgagga
      181 agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga
      241 ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc cggacccacc
      301 ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc cgctgcccca
      361 aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac tagtgagacg
      421 tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg ggaacttcct
      481 gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag ccggttggcg
      541 cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc gccaagtgcc
      601 cagcggggct gctaaagcgc atgctccaga ctgccttggg aaaagcgcct cccctacccg
      661 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      721 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      781 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      841 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      901 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      961 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1021 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     1081 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     1141 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     1201 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     1261 gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag agccgccacc
     1321 atggataaaa agtattctat tggtttagcc atcggcacta attccgttgg atgggctgtc
     1381 ataaccgatg aatacaaagt accttcaaag aaatttaagg tgttggggaa cacagaccgt
     1441 cattcgatta aaaagaatct tatcggtgcc ctcctattcg atagtggcga aacggcagag
     1501 gcgactcgcc tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa ccgaatatgt
     1561 tacttacaag aaatttttag caatgagatg gccaaagttg acgattcttt ctttcaccgt
     1621 ttggaagagt ccttccttgt cgaagaggac aagaaacatg aacggcaccc catctttgga
     1681 aacatagtag atgaggtggc atatcatgaa aagtacccaa cgatttatca cctcagaaaa
     1741 aagctagttg actcaactga taaagcggac ctgaggttaa tctacttggc tcttgcccat
     1801 atgataaagt tccgtgggca ctttctcatt gagggtgatc taaatccgga caactcggat
     1861 gtcgacaaac tgttcatcca gttagtacaa acctataatc agttgtttga agagaaccct
     1921 ataaatgcaa gtggcgtgga tgcgaaggct attcttagcg cccgcctctc taaatcccga
     1981 cggctagaaa acctgatcgc acaattaccc ggagagaaga aaaatgggtt gttcggtaac
     2041 cttatagcgc tctcactagg cctgacacca aattttaagt cgaacttcga cttagctgaa
     2101 gatgccaaat tgcagcttag taaggacacg tacgatgacg atctcgacaa tctactggca
     2161 caaattggag atcagtatgc ggacttattt ttggctgcca aaaaccttag cgatgcaatc
     2221 ctcctatctg acatactgag agttaatact gagattacca aggcgccgtt atccgcttca
     2281 atgatcaaaa ggtacgatga acatcaccaa gacttgacac ttctcaaggc cctagtccgt
     2341 cagcaactgc ctgagaaata taaggaaata ttctttgatc agtcgaaaaa cgggtacgca
     2401 ggttatattg acggcggagc gagtcaagag gaattctaca agtttatcaa acccatatta
     2461 gagaagatgg atgggacgga agagttgctt gtaaaactca atcgcgaaga tctactgcga
     2521 aagcagcgga ctttcgacaa cggtagcatt ccacatcaaa tccacttagg cgaattgcat
     2581 gctatactta gaaggcagga ggatttttat ccgttcctca aagacaatcg tgaaaagatt
     2641 gagaaaatcc taacctttcg cataccttac tatgtgggac ccctggcccg agggaactct
     2701 cggttcgcat ggatgacaag aaagtccgaa gaaacgatta ctccatggaa ttttgaggaa
     2761 gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccaa ctttgacaag
     2821 aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg
     2881 tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta
     2941 agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca
     3001 gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc tgtcgagatc
     3061 tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct cctaaagata
     3121 attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga agatatagtg
     3181 ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa aacatacgct
     3241 cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac gggctgggga
     3301 cgattgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa aactattctc
     3361 gattttctaa agagcgacgg cttcgccaat aggaacttta tgcagctgat ccatgatgac
     3421 tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg ggactcattg
     3481 cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat actccagaca
     3541 gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga aaacattgta
     3601 atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag tcgagagcgg
     3661 atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa ggagcatcct
     3721 gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca aaatggaagg
     3781 gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga cgtcgatgcc
     3841 attgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct tacacgctcg
     3901 gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa gaaaatgaag
     3961 aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt cgataactta
     4021 actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat taaacgtcag
     4081 ctcgtggaaa cccgccaaat cacaaagcat gttgcacaga tactagattc ccgaatgaat
     4141 acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac tttaaagtca
     4201 aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga gataaataac
     4261 taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact cattaagaaa
     4321 tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga cgtccgtaag
     4381 atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt cttttattct
     4441 aacattatga atttctttaa gacggaaatc actctggcaa acggagagat acgcaaacga
     4501 cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg ccgggacttc
     4561 gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa aactgaggtg
     4621 cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga taagctcatc
     4681 gctcgtaaaa aggactggga cccgaaaaag tacggtggct tcgatagccc tacagttgcc
     4741 tattctgtcc tagtagtggc aaaagttgag aagggaaaat ccaagaaact gaagtcagtc
     4801 aaagaattat tggggataac gattatggag cgctcgtctt ttgaaaagaa ccccatcgac
     4861 ttccttgagg cgaaaggtta caaggaagta aaaaaggatc tcataattaa actaccaaag
     4921 tatagtctgt ttgagttaga aaatggccga aaacggatgt tggctagcgc cggagagctt
     4981 caaaagggga acgaactcgc actaccgtct aaatacgtga atttcctgta tttagcgtcc
     5041 cattacgaga agttgaaagg ttcacctgaa gataacgaac agaagcaact ttttgttgag
     5101 cagcacaaac attatctcga cgaaatcata gagcaaattt cggaattcag taagagagtc
     5161 atcctagctg atgccaatct ggacaaagta ttaagcgcat acaacaagca cagggataaa
     5221 cccatacgtg agcaggcgga aaatattatc catttgttta ctcttaccaa cctcggcgct
     5281 ccagccgcat tcaagtattt tgacacaacg atagatcgca aacgatacac ttctaccaag
     5341 gaggtgctag acgcgacact gattcaccaa tccatcacgg gattatatga aactcggata
     5401 gatttgtcac agcttggggg tgacggatcc cccaagaaga agaggaaagt ctcgagcgac
     5461 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     5521 aaggctgcag gaggcggagg tagcaaccac gaccaggaat ttgaccctcc aaaggtttac
     5581 ccacctgtcc cagctgagaa gaggaagccc atccgggtgc tgtctctctt tgatggaatc
     5641 gctacagggc tcctggtgct gaaggacttg ggcattcagg tggaccgcta cattgcctcg
     5701 gaggtgtgtg aggactccat cacggtgggc atggtgcggc accaggggaa gatcatgtac
     5761 gtcggggacg tccgcagcgt cacacagaag catatccagg agtggggccc attcgatctg
     5821 gtgattgggg gcagtccctg caatgacctc tccatcgtca accctgctcg caagggcctc
     5881 tacgagggca ctggccggct cttctttgag ttctaccgcc tcctgcatga tgcgcggccc
     5941 aaggagggag atgatcgccc cttcttctgg ctctttgaga atgtggtggc catgggcgtt
     6001 agtgacaaga gggacatctc gcgatttctc gagtccaacc ctgtgatgat tgatgccaaa
     6061 gaagtgtcag ctgcacacag ggcccgctac ttctggggta accttcccgg tatgaacagg
     6121 ccgttggcat ccactgtgaa tgataagctg gagctgcagg agtgtctgga gcatggcagg
     6181 atagccaagt tcagcaaagt gaggaccatt actacgaggt caaactccat aaagcagggc
     6241 aaagaccagc attttcctgt cttcatgaat gagaaagagg acatcttatg gtgcactgaa
     6301 atggaaaggg tatttggttt cccagtccac tatactgacg tctccaacat gagccgcttg
     6361 gcgaggcaga gactgctggg ccggtcatgg agcgtgccag tcatccgcca cctcttcgct
     6421 ccgctgaagg agtattgaaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     6481 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     6541 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     6601 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     6661 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctcgatac
     6721 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6781 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctagg
     6841 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6901 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6961 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     7021 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     7081 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7141 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7201 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7261 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7321 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7381 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7441 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7501 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7561 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7621 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7681 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7741 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7801 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7861 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7921 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7981 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8041 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8101 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8161 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8221 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8281 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8341 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8401 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8461 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8521 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8581 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8641 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8701 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8761 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8821 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8881 gcacatttcc ccgaaaagtg ccacctgacg tcgacgggta tacagacatg ataagataca
     8941 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     9001 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg
     9061 gcgaagaact ctcggcatct actctattcc tttgccctcg gacgagtgct ggggcgtcgg
     9121 tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc gcttctgcgg
     9181 gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc gcatcgaccc
     9241 tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag ttggtcaaga
     9301 ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg caagctccgg atgcctccgc
     9361 tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa gaagatgttg
     9421 gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac cgctgttatg
     9481 cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag gtgccggact
     9541 tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac ggacgcactg
     9601 acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat cgcgcatatg
     9661 aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc
     9721 gtctggctaa gatcggccgc agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca
     9781 gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgcacg gcgggagatg
     9841 caataggtca ggctctcgct gaattcccca atgtcaagca cttccggaat cgggagcgcg
     9901 gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc gcagctattt
     9961 acccgca
//
